Agriculturally Important Microbes For Su
Agriculturally Important Microbes For Su
Agriculturally Important Microbes For Su
Agriculturally
Important Microbes
for Sustainable
Agriculture
Volume I: Plant-soil-microbe nexus
Agriculturally Important Microbes
for Sustainable Agriculture
Vijay Singh Meena • Pankaj Kumar Mishra
Jaideep Kumar Bisht • Arunava Pattanayak
Editors
Agriculturally Important
Microbes for Sustainable
Agriculture
Volume I: Plant-soil-microbe nexus
Editors
Vijay Singh Meena Pankaj Kumar Mishra
ICAR-Vivekananda Institute of Hill ICAR-Vivekananda Institute of Hill
Agriculture Agriculture
Almora, Uttarakhand, India Almora, Uttarakhand, India
v
vi Contents
vii
Part I
Role of Soil Microbe Interaction
Plant Beneficial Rhizospheric Microbes
(PBRMs): Prospects for Increasing 1
Productivity and Sustaining
the Resilience of Soil Fertility
Abstract
The efficient microorganisms in the vicinity of plant roots that exert positive
effects on plant growth are known as plant beneficial rhizospheric microbes
(PBRMs). One of the salient features for the effectiveness of PBRMs is their
ability to proliferate at hosts’ rhizosphere, rhizoplane or the root interior. They
are directly or indirectly involved in plant growth promotion and development by
releasing particular compounds, increasing the uptake of certain nutrients from
the soil and lessening or preventing the plants from deleterious pathogens.
Indirect plant growth promotion is achieved by the inhibition of the detrimental
effects of phytopathogenic organisms, which can be obtained by the production
of siderophores, i.e. metal-chelating agent. Several bacterial species also have
been reported in biocontrol of soil-borne phytopathogens and the synthesis
of antibiotics. Production of hydrogen cyanide (HCN) and/or fungal cell wall
degrading enzymes (e.g. chitinase and β-1, 3-glucanase) help PBRMs to sup-
A. Sarkar (*)
ICAR-Indian Institute of Water Management, Bhubaneswar 751023, Odisha, India
ICAR-Indian Agricultural Research Institute, Pusa Campus, New Delhi 110012, India
e-mail: asiari2012@gmail.com
M. Saha
Department of Agricultural Chemistry and Soil Science, Faculty of Agriculture, Bidhan
Chandra KrishiViswavidyalaya, Mohanpur 741252, West Bengal, India
Department of Soil Science and Agricultural Chemistry, Institute of Agricultural Sciences,
Banaras Hindu University, Varanasi 221005, Uttar Pradesh, India
e-mail: madhumonti2609@gmail.com
V.S. Meena
ICAR-Vivekananda Institute of Hill Agriculture, Almora 263601, Uttarakhand, India
Department of Soil Science and Agricultural Chemistry, Institute of Agricultural Sciences,
Banaras Hindu University, Varanasi 221005, Uttar Pradesh, India
e-mail: vijay.meena@icar.gov.in
press the effect of phytopathogens. Direct plant growth promotion involves sym-
biotic and non-symbiotic PBRMs which function through production of plant
hormones such as auxins, gibberellins, ethylene, cytokinins and abscisic acid.
Some of the PBRMs are functioning as a sink for 1-aminocyclopropane-1-
carboxylate (ACC), the immediate precursor of ethylene in higher plants, by
hydrolyzing it into α-ketobutyrate and ammonia and in this way promoting
root growth by lowering indigenous ethylene levels in the rhizospheric environ-
ment. PBRMs also help in solubilization of nutrient-bearing minerals (indige-
nous sources of phosphates, potash and other nutrients), enhance resistance to
plant stress, stabilize soil aggregates and improve soil structure and organic mat-
ter content. PBRMs could able to retain more soil organic N and other nutrients
in the plant-soil system, which can reduce the need for fertilizer N and P and help
in nutrient acquisition. Evidently, PBRMs maintain enormous prospects in
advanced and sustainable plant production, including, improved plant tolerance
to stress, better uptake of plant nutrient from soils and reduced application of
chemical inputs. Besides, PBRMs in the soil bear a peculiar relation to soil fertil-
ity. Various investigations have documented the increased health and productiv-
ity of different plant species by the application of plant beneficial rhizospheric
microorganisms under both the normal and abiotic-stressed conditions. Attempts
should be directed towards maximizing the identified benefits of PBRMs in all
developing countries. If the benefits of PBRMs in crop production can be
maximized, this will certainly help to fight against hunger. This chapter dis-
cusses the major functions of PBRMs in broad terms, but efforts were made to
present specific usage of PBRMs to enhance plant nutrient uptake, for better
plant response to environmental stress, and unexplored potentials in developing
countries.
Keywords
Rhizosphere • PBRMs • Stress resistance • Nutrient acquisition • Sustainability
1.1 Introduction
Plants are the sources of food, oxygen, fuel, fibre, medicine and residence for every
animal on this planet Earth. Whereas, these plants uptake their own requirements
mainly from soil through its root system. Thus, root-soil interaction is running
simultaneously from the beginning and evolution of life, where they assemble ben-
efits from their symbiotic association with each other (Meena et al. 2013a, 2016a;
Bahadur et al. 2014; Maurya et al. 2014; Jat et al. 2015; Ahmad et al. 2016; Parewa
et al. 2014; Kumar et al. 2015, 2016b). This root-soil-associated phase is known as
rhizosphere (rhiza = root, sphere = area of influence).The rhizosphere is the narrow
region of soil that is directly influenced by plant roots in association with root hairs,
plant-produced materials and concerted soil microorganisms (Hartmann et al.
2009). It may expand up to 2–3 mm from the root surface, which can be
1 Plant Beneficial Rhizospheric Microbes (PBRMs): Prospects for Increasing… 5
Fig. 1.1 Distribution of microbes in rhizospheric zoo (Coyne 1999; Mendes et al. 2013)
Priyadharsini and Muthukumar 2016; Meena et al. 2015f, 2016b; Jaiswal et al.
2016; Jha and Subramanian 2016; Kumar et al. 2016a).
As seeds germinate and roots grow throughout the soil, they secrete organic
metabolites which supply the driving force for the activation of microbial popula-
tions in a zone which includes plant root and the adjacent soil in a few millimetre
(mm) of thickness. This is referred as the rhizosphere effect (Morgan et al. 2011)
and considered as the qualitative estimation of the degree or extent of plant growth
that are affected by those microorganisms, while R:S ratio is used as quantitative
measure. These plant beneficial microbes (PBMs) are very indispensable in biogeo-
chemical cycles which are the determinants of plant health and soil fertility and
have been used for crop production for decades. Generally, the microbe functions in
different ways such as secreting regulatory chemical compounds for the plants,
facilitating resource acquisition of certain nutrients (N, P, K, Fe, S, etc.) from the
soil and reducing or impeding the plants from the inhibitory effects of various
pathogens in the forms of a biocontrol agent (Kumar et al. 2017; Raghavendra et al.
2016; Zahedi 2016; Meena et al. 2015a, b; Das and Pradhan 2016; Dominguez-
Nunez et al. 2016; Dotaniya et al. 2016).
Evidently, these rhizospheric microbes hold infinite potential in advanced and
sustainable plant production, including reinforced plant tolerance to stress, modu-
lated plant hormone levels and limited use of chemical inputs. Regular use of enor-
mous amounts of chemical inputs is causing unsuitable environment. Now, try to
better understand the function of inoculants as biofertilizers in nutrient absorption,
and plant response to environmental stress is more enthralling. The plant beneficial
rhizobacteria may reduce the worldwide dependence on precarious agricultural
chemicals which weaken the agroecosystems. This chapter emphasizes the compre-
hension of the rhizosphere and plant beneficial rhizospheric microorganisms under
the present aspects (Rawat et al. 2016; Yasin et al. 2016; Meena et al. 2015e, 2016c,
d; Saha et al. 2016b; Yadav and Sidhu 2016; Teotia et al. 2016; Bahadur et al.
2016b).
The chemicals which are released by the roots within the soils are generally known
as root exudates. Then the root products contributed to the surrounding soil are
generally called as rhizodeposits. Root exudates, a fraction of rhizodeposition, are
abundant in carbon and energy sources that govern the occurrence and growth of the
microbes in the rhizosphere. Other fractions of rhizodeposition are lysates, muci-
lage, secretions and dead cell materials that may take part in plant development
(Sommers et al. 2004; Dardanelli et al. 2010).
It is learned that both the qualitative and quantitative formulations of root exu-
dates are influenced by different environmental factors such as pH, soil type, oxy-
gen status, light intensity, soil temperature, nutrient accessibility and the presence
1 Plant Beneficial Rhizospheric Microbes (PBRMs): Prospects for Increasing… 7
Table 1.1 Various compounds in root exudates (rhizodeposits) of different plant species
Amino acids α-Alanine, β-alanine, asparagines, aspartate, cysteine, cystine,
glutamate, glycine, isoleucine, leucine, lysine, methionine,
serine, threonine, proline, valine, tryptophan, ornithine, histidine,
arginine, homoserine, phenylalanine, aminobutyric acid,
α-aminoadipic acid
Enzymes Acid/alkaline-phosphatase, invertase, amylase, protease
Inorganic ions and gaseous HCO3−,OH−, H+, CO2, H2
molecules
Organic acids Citric acid, oxalic acid, malic acid, fumaric acid, succinic acid,
acetic acid, butyric acid, valeric acid, glycolic acid, piscidic acid,
formic acid, aconitic acid, lactic acid, pyruvic acid, glutaric acid,
malonic acid, tetronic acid, aldonic acid, erythronic acid
Purines/nucleosides Adenine, guanine, cytidine, uridine
Sugars Glucose, fructose, galactose, ribose, xylose, rhamnose,
arabinose, desoxyribose, oligosaccharides, raffinose, maltose
Vitamins Biotin, thiamin, pantothenate, riboflavin, niacin
Adapted from Dakora and Phillips 2002
2013; Saha et al. 2016b; Verma et al. 2014, 2015c; Meena et al. 2014a, 2016e;
Sharma et al. 2016).
Fig. 1.2 Schematic diagram representing the direct and indirect effects of plant beneficial rhizo-
spheric microbes (PBRMs)
1 Plant Beneficial Rhizospheric Microbes (PBRMs): Prospects for Increasing… 9
much convenient in making unavailable form of soil nutrients into readily available
form by exploring the large volumes of soil (Velazquez et al. 2016; Meena et al.
2014b, 2015c; Sindhu et al. 2016; Bahadur et al. 2016a).
Their hyphae developed superficially from the roots ranging from a few centime-
tres (cm) up to several meters in the soil. As a consequences, it increases the effec-
tive absorbing surface area of the host plant root by as much as ten times, resulting
in increased absorption of static nutrients such as phosphorus, zinc, copper, etc. in
the soil by ~60 times.
In India, inoculation with VAM fungi (Glomus mosseae, G. fasciculatum,
Acaulospora laevis and Gigaspora gilmorei) resulted increased plant productivity
(Kumar et al. 2009). These PSB and PSF (fungi) correspondingly represent ~1–50%
and 0.1–0.5% of the total population of microorganisms present in the soil (Chabot
et al. 1993; Khan et al. 2009).
The PSB solubilize inorganic soil phosphates, being Ca3(PO4)2, FePO4 and
AlPO4, through the production of organic acids, siderophores and hydroxyl and
carboxyl groups that chelate the cation bound to phosphate and the latter being con-
verted into soluble forms (Sharma et al. 2013). Bacterial isolates belonging to gen-
era Enterobacter, Pantoea and Klebsiella solubilize Ca3(PO4)2 to a greater extent
than FePO4 and AlPO4 (Chung et al. 2005). Hence, microorganisms play an impor-
tant role in the soil phosphorus cycle by mediating phosphorus availability to plants,
increasing the capacity of plants to acquire phosphorus from the P-deficient soil
directly by solubilizing inorganic phosphorus and facilitating their mobilization
through the enlargement of root surface area (Richardson and Simpson 2011).
India is blessed with sufficient supply of potassium (K) in soil, though the K reserve
is limited, which results to a huge foreign exchange to import K fertilizers. In this con-
text utilization of native sources such as K-bearing minerals (K-feldspar, phlogopite,
biotite, muscovite and green sands), waste mica as enriched compost (Nishanth and
Biswas 2008) and bio-intervention of these K-bearing minerals could be a viable and
alternative technology to solubilize insoluble K into plant available pool.
The usefulness of K-bearing minerals as K fertilizers in agriculture is less due to
their limited accessibility. Some efficient microorganisms play significant induction
in K solubilization and K release from K-bearing minerals (Reitemeir 1951); those
microorganisms are commonly described as potassium-solubilizing bacteria (KSB),
potassium-solubilizing rhizobacteria (KSR) and potassium-dissolving bacteria
(KDB). A broad group of bacteria named Pseudomonas sp., Acidithiobacillus fer-
rooxidans, Bacillus circulans, B. mucilaginosus, Burkholderia, B. edaphicus and
Paenibacillus sp. have been reported to release potassium in available form from
potassium-bearing minerals in soils (Liu et al. 2012; Bagyalakshmi et al. 2012;
Saiyad et al. 2015; Verma et al. 2015a).
As per the finding of Hutchens et al. (2003), they reported that some specific
bacteria generate mucilage like exopolysaccharides which form a cover on all the
sides of the bacterial cell and invade the silicate minerals chelating with silicon and
hence facilitate K from those silicate structure. It was found that various organic
ligands include exudates, extracellular enzymes, metabolic by-products and che-
lates, and both light and compound organic acids (oxalic, citric, gluconic acids) and
1 Plant Beneficial Rhizospheric Microbes (PBRMs): Prospects for Increasing… 11
1.3.2 B
iocontrol Activities of Plant Beneficial Rhizospheric
Microbes
Plant growth and development can be obtained indirectly via biocontrol activity in
opposition to plant pathogens. Representatives of the bacterial genera Bacillus,
Pseudomonas, Serratia, Stenotrophomonas and Streptomyces and the fungal genera
Ampelomyces, Coniothyrium and Trichoderma are well-defined microorganisms
which are responsible for resistant action and include hindrance of the pathogen by
the production of antibiotics, toxins and surface-active compounds (biosurfactants);
competition for nutrients, minerals and colonization sites (Table 1.3); and a
1 Plant Beneficial Rhizospheric Microbes (PBRMs): Prospects for Increasing… 13
Table 1.3 Different biocontrol agents produced by PBRM that control disease, pathogen and
insect at different crops
Disease/pathogen/insect Crops PBRM
Myzus persicae (Sulzer) Bell pepper Bacillus amyloliquefaciens
Fusarium avenaceum Chickpea Enterobacter sp.
Cotton aphids Cucumber Bacillus sp.
Root rot Mung bean Pseudomonas aeruginosa
Myzus persicae Pepper Bacillus licheniformis
Myzus persicae Pepper Bacillus subtilis G803
Rhizosphere fungi Prunus cerasifera L. Azospirillum brasilense
Myzus persicae Red pepper Bacillus cereus MJ-1
Fungal disease Sesame Paenibacillus polymyxa E681
Blue mould Tobacco Bacillus pumilus SE 34
Rhizoctonia bataticola Tobacco Pseudomonas sp.
Tobacco necrosis virus Tobacco Pseudomonas fluorescens
Tomato mottle virus Tomato Bacillus amyloliquefaciens
Acyrthosiphon kondoi White clover Medicago Pseudomonas sp.
Adopted from Bhattacharyya and Jha 2012
procedure that grows extracellular cell wall deteriorating enzymes such as chitinase
and β-1,3-glucanase (Whipps 2001; Compant et al. 2005; Haas and Defago 2005).
Effective plant growth acceleration and biological control of plant maladies by
rhizobacteria include one or more procedures (Fig. 1.3), which involve the genera-
tion of phytohormones, parasitism, antibiosis, competition for niches and nutrients
and induced host defence (Lugtenberg and Kamilova 2004; Adesemoye et al. 2009).
In general, niche ejection, induced systemic resistance (ISR) and production of
antifungal metabolites are the cardinal functions of biocontrol activity in
PBRMs (Lugtenberg and Kamilova 2009). Many rhizobacteria have been
described to grow antifungal metabolites such as HCN, phenazines, pyrrolnitrin,
2,4-diacetylphloroglucinol, pyoluteorin, viscosinamide and tensin (Bhattacharyya
and Jha 2012). The ISR is the circumstance in which the mutual cooperation of
some rhizobacteria with the host root protects the plants from some pathogenic
bacteria, viruses and fungi (Lugtenberg and Kamilova 2009). Several strains from
Pseudomonas, Bacillus and Azospirillum genera are the dominant group of that
have been characterized for generating the ISR response.
Furthermore, ISR comprises jasmonate and ethylene signalling inside the
plants, and these hormones activate resistance capacity of the host plant against a
range of plant pathogens (Glick 2012). Many discrete bacterial ingredients induce
ISR, such as lipopolysaccharides (LPS), siderophores, flagella, cyclic lipopeptides,
2, 4-diacetylphloroglucinol, homoserine lactones and volatiles like acetoin and
2,3-butanediol (Lugtenberg and Kamilova 2009). With the development of materi-
alistic PGPR in India, the sheath blight of rice due to Rhizoctonia solani was alleviated
over the control ~38%, and grain yields were significantly increased (~3901
and 1938 kg ha−1) over control (~2690 and 1550 kg ha−1) (Kumar et al. 2009). The
14 A. Sarkar et al.
Fig. 1.3 Nutrient acquisition and induced systemic resistance (ISR) against draught, salt and fer-
tility stress by different efficient PBRMs (Coyne 1999; Yang et al. 2009). The dashed arrows rep-
resent PBRMs secreted compounds; solid arrows indicate the components affected by PBRMs
secreted compounds (VAM vesicular arbuscular mycorrhiza, ABA abscisic acid, ROS reactive oxy-
gen species, ACC 1-aminocyclopropane-1-carboxylate, PBRM plant beneficial rhizospheric
microbes, IAA indole acetic acid)
‘Stress’ in plants can be defined as any external factor that negatively influences
plant growth, productivity, reproductive capacity or survival. This includes a wide
range of factors which can be broadly divided into two main categories: abiotic or
environmental stress factors and biotic or biological stress factors. Biotic stress can
be defined as the stress which is induced by living organisms, such as other plants,
weeds, fungi, bacteria, different faunal groups, etc., whereas abiotic stress is the
result of nonliving stressors, such as high winds, extreme temperature and rainfall,
drought and unwanted and devastated natural calamities, like cyclone, tornado, flash
flood, etc.
1 Plant Beneficial Rhizospheric Microbes (PBRMs): Prospects for Increasing… 15
Under these abiotic and biotic stresses, endogenous ethylene production is sig-
nificantly increased which adversely affects the growth of the roots and as a whole
the growth of the plant. In this concern, some distinct soil microorganisms produce
some growth factors and metabolic products that positively influence plant metabo-
lism which is not directly associated with pest or pathogen resistance (biotic stress).
A number of mechanisms have been investigated aiming to reduce the levels of
ethylene in plants. One of these mechanisms involves the activity of the enzyme
1-aminocyclopropane-1-carboxylase (ACC) produced by soil bacteria and degrades
an ethylene precursor that, in turn, depresses the plant’s stress response to a variety
of biotic and abiotic stress factors (Farajzadeh et al. 2012). Soil bacteria also pro-
duce the auxin, indole acetic acid (IAA) which influences plant physiology, often
resulting in enhanced root growth which stimulates plant growth. Naturally, micro-
bial metabolites that positively influence plant vigour also impact plant resistance to
pests and pathogens (Kim et al. 2011).
The AA’s rhizosphere is the immediate adjacent zone of plant root and a nutrient-
rich environment due to adequate supply of sugars, amino acids, organic acids, fla-
vonoids, iso-flavonoids, phytohormones and enzymes; it is also the most vulnerable
environment during activity of stressors and stress conditions. Thus, controlling and
sustaining rhizosphere in its healthy manner are a major challenge for agricultural
science societies. The applications of beneficial microbes in agricultural production
systems were introduced long time ago, and there was enhancing manifestation that
beneficial microbes can increase plants’ tolerance to unfavourable environmental
difficulties, which comprises salt stress (Egamberdiyeva and Islam 2008), drought
stress (Zahir et al. 2008), weed infestation (Babalola 2010), nutrient deficiency and
heavy metal contaminations (Sheng 2005).
The term ‘induced systemic tolerance’ (Fig. 1.3) has been applied to define the
potentiality of PGPR to induce tolerance to salt and drought conditions (Yang et al.
2009). In this concern Rhizobium trifolii inoculated with Trifolium alexandrinum
(berseem) has shown excessive biomass and large number of nodulation under
salinity stress condition (Hussain et al. 2002; Antoun and Prevost 2005). Marulanda
et al. (2009) rightly observed that the combination of Pseudomonas putida or
Bacillus megaterium and AM fungi had been efficiently mitigating the drought
stress. Photosynthetic ability and the anti-oxidative activity of rice plants liable to
drought stress had shown to increase after inoculation with the arbuscular mycor-
rhiza (Ruiz-Sanchez et al. 2010).
The auspicious effects of mycorrhizae have also been described under both the
saline and drought stress conditions (Aroca et al. 2013). Not only mycorrhizae but
also identification of diverse salt-tolerant rhizobacteria has displayed favourable
interactions with plants under stressed conditions. These PGPR (Rhizobium,
Azospirillum, Pseudomonas, Flavobacterium, Arthrobacter and Bacillus) exert
osmoregulation, resistance to starvation, oligotrophic and endogenous metabolism
and useful metabolic methods to acclimatize under arid and brackish environments
(Lugtenberg et al. 2001; Egamberdiyeva and Islam 2008). When plants are illumi-
nated to the stress situations, they may increase ethylene inside the cell and cause
plant damage (Argueso et al. 2009).
16 A. Sarkar et al.
nutrients from the surrounding soils. Indole-3-acetic acid (IAA) is a commonly pro-
duced phytohormone by growth-promoting rhizospheric microorganisms and which
is believed to be associated with cell division, cell enlargement and root initiation
(Salisbury 1996). Auxin is a group of plant hormones significant in the advance-
ment of lateral root formation. Enhanced lateral root leads to an increased ability to
absorb the essential nutrients for the plant.
Auxins are mostly secreted by Azospirillum, and it is commonly described that
their formation, rather than the nitrogen fixation, is the main feature responsible for
the stimulation of lateral root growth and, hence, advanced plant development
(Bloemberg and Lugtenberg 2001). Other groups of plant hormones include gib-
berellins and cytokinins, both of which regulate shoot development; howsoever
their effects on root enlargement are not well researched (Fig. 1.3). They meet
important functions for plant growth and health by several processes. Immediate
plant growth promotion occurs either from enhanced nutrient availability or from
hormonal incitement. Variant methods are engaged in the inhibition of plant patho-
gens, which is mostly indirectly related with plant growth.
Plant growth and productivity are greatly affected by the mutual associations
between plant roots and the surrounding soil, including the microbial communities.
These microbes are either autochthonous or applied as crop inoculation such as
Azospirillum (Cassan and Garcia de Salamone 2008), Bacillus (Jacobsen et al.
2004), Pseudomonas (Loper 1988), Rhizobium (Long 2001), Serratia (De
18 A. Sarkar et al.
1.3.5 R
ole of Rhizospheric Microbes in Soil Fertility
and Sustainability
‘Zero Hunger Challenge’ is the major goal of human efforts. This goal can be
achieved by sustainable increment in the arena of agricultural productivity to satisfy
the demands of abruptly spreading human population. Growing more and more
foods from less and less farm holdings is a demand feature of our economy. Not
only we have touched the limits of land ready for cultivation, but agricultural mar-
gin is also narrowing at the proportion of a million acres per annum due to enlarge-
ment of our cities, industrial areas and highways construction. Then we should
consider our soil with great concern. Microorganisms in the soil show a unique
relation to soil fertility. The presence of rhizosperic microbial communities in asso-
ciation with PBRMs has pivotal role to prolong soil function, in both natural and
maintained agricultural soils (Bharti et al. 2016) (Table 1.5).
Soil organisms exceed soil fertility by doing a number of functions that are
advantageous for plants. This chapter assesses five of these functions of PBRMs:
1. PBRMs can accelerate the nutrient release from native source of nutrient in soil
and applied organic residues. Phosphate-solubilizing PBRMs and potassium-
solubilizing PBRMs accelerate the weathering reactions of P- and K-bearing
minerals, especially when in direct contact with mineral surfaces either by pro-
duction of organic acids or formation of biofilms or may take part in nutrient
mobilization by proliferation of root length as mycorrhizal fungi (Basak and
Biswas 2009; Saha et al. 2016a; Hodge 2017).
2. In the symbiotic association, Rhizobia or Bradyrhizobia fix atmospheric nitro-
gen gas and make it available to the leguminous plants. Free-living (Enterobacter,
Azotobacter) and associative (Azospirillum) nitrogen-fixing rhizobacteria, nev-
ertheless, absorb this gas and transform it into cell protein. When the cells waste
away, other microbes invade the protein and convert the nitrogenous components
to ammonium, so which becomes readily available for the plants.
3. The process of ‘the priming effect’ has described as early as 1926 but still poorly
understood. Positive priming (extra decomposition of SOM) or negative priming
(reduction of SOM decomposition) is depending on the metabolic activity of
PBRMs (van der Wal and de Boer 2017). In terrestrial ecosystems, large quanti-
ties of carbon are held in SOM ‘the key to soil fertility’ and altered by soil
microbes that supply SOC and organically bounded nutrients to plants.
4. Bioremediation of agricultural toxic chemical inputs (pesticides) in soil is mostly
acted by microorganisms (Zhuang et al. 2007). Some PBRMs having enzymatic
activity in soil, which can disintegrate agricultural pesticides or other toxic
1 Plant Beneficial Rhizospheric Microbes (PBRMs): Prospects for Increasing… 19
Table 1.5 Different PBRMs of vegetable and food grains and their activities
Host plants PBRM inoculants PBRM abilities
Vegetables Brinjal Azotobacter spp., Azospirillum Biofertilizer
spp.
Pseudomonas fluorescens Biocontrol against bacterial
wilt
Tomato Bacillus subtilis, B. cereus, Induced systemic resistance
Arthrobacter spp., Trichoderma and biocontrol against late
viride, Pseudomonas blight
fluorescens, P. putida, P.
marginalis, P. syringae
Potato Bacillus subtilis, B. cereus, Biofertilizer, induced systemic
Azotobacter spp. resistance against late blight
Broccoli Bacillus cereus, Brevibacillus Biofertilizer
reuszeri, Rhizobium rubi
Cabbage Bacillus subtilis, B. Improved nutrient uptake,
megaterium, Pantoea induced systemic resistance
agglomerans
Okra Bacillus firmus, B. subtilis Induced systemic resistance
against root knot nematode
Radish Pseudomonas fluorescens, Induced systemic resistance
Bacillus subtilis
Chilli Bacillus spp., Arthrobacter and Induced systemic resistance
Serratia spp. against anthracnose disease of
chilli (Colletotrichum
gloeosporioides) and
endophytic bacteria
Spinach Bacillus megaterium Induced systemic resistance
B. subtilis against Rhizoctonia, Pythium,
B. licheniformis Fusarium and Phytopthora
B. cereus
B. subtilis
Paenibacillus polymyxa
Pseudomonas putida
Cucumber Bacillus pumilus Biocontrol against angular leaf
Bacillus spp. spot (Pseudomonas syringae)
Biocontrol against root rot
(Pythium sp.)
Food grains Rice Pseudomonas fluorescence Biocontrol against
Aur6, Magnaporthe grisea
Chryseobacterium balustinum, Salinity
BGA N fixation
Wheat Bacillus circulans, Glomas Biofertilization
spp., Cladosporium herbarum, Salinity controller
Calothrix sp., Anabaena sp.
Arthrobacter sp., Bacillus
subtilis
(continued)
20 A. Sarkar et al.
Table 1.5 (continued)
Host plants PBRM inoculants PBRM abilities
Maize Serratia liquefaciens, Bacillus Biofertilization
sp. Biocontrol to root pathogens
Pseudomonas spp. Phytostimulation
Azospirillum brasilense,
Bradyrhizobium japonicum
Soybean Bradyrhizobium japonicum, Biofertilization enhancer
Bacillus subtilis, Bacillus Phytostimulation
thuringiencis
Pseudomonas flaviporus,
Bradyrhizobium japonicum
Bean Rhizobium tropici, Azotobacter Biofertilization enhancer (N
brasilence, Glomus sinuosum, and P)
Glomus intraradices, Biocontrol against Rhizoctonia
Gigaspora albida, solani
Pseudomonas fluorescens
Adopted from Choudhary and Johri 2009; Zaidi et al. 2015; Montano et al. 2014
Being an inherent part of soil and as most of the soil function is microbial
imparted, soil organisms play an essential role in conserving soil health and sustain-
able crop production even after decades. The main target in the upcoming decades
would be on secured and eco-friendly approaches by accomplishing the beneficial
microorganisms in sustainable crop production. Commonly, there are divergent in
naturally occurring microorganisms, whose inoculation to the soil ecological envi-
ronment improves soil physicochemical properties, soil microbial proliferation, soil
quality, plant growth and crop productivity (Morris and Blackwood 2015).
microorganisms in crop production can be widen, this will certainly take an action
against hunger. The production performance of a specific rhizospheric microorgan-
ism may be further enlarged with the development and adaptation according to the
prevalent soil conditions. Subsequently, they are supposed to partly substitute the
chemical fertilizers, pesticides and synthetic growth regulators which have numer-
ous side effects to sustainable agriculture (Verma et al. 2015c). Notwithstanding the
fact that a lot of associative and endophytic bacteria have indicated plant growth-
promoting properties at the point of laboratory and greenhouse level, but these bac-
teria have dimed to show their consistency under natural conditions. Accordingly, a
challenge is affixed for standard optimization of the applied efficient PGPR
(PBRMs) strains and the amount of fertilizers to be added to get the maximum out-
put. One of the major challenges encompasses the specification of plant genotype,
age and suitable associative bacteria. Considering this suitability, it would help to
increase productivity by adopting the specific strain for inoculation.
1.5 Conclusions
References
Adesemoye A, Torbert H, Kloepper JW (2009) Plant growth-promoting rhizo-bacteria allow
reduced application rates of chemical fertilizers. Microbial Ecol 58:921–929
Ahemad M, Khan MS (2012) Evaluation of plant growth promoting activities of rhizobacterium
Pseudomonas putida under herbicide-stress. Ann Microbiol 62:1531–1540
22 A. Sarkar et al.
Ahmad M, Nadeem SM, Naveed M, Zahir ZA (2016) Potassium-solubilizing bacteria and their
application in agriculture. In: Meena VS, Maurya BR, Verma JP, Meena RS (eds) Potassium
solubilizing microorganisms for sustainable agriculture. Springer, India, pp 293–313.
doi:10.1007/978-81-322-2776-2_21
Alves BJR, Boddey RM, Urquiaga S (2004) The success of BNF in soybean in Brazil. Plant Soil
252:1–9
Antoun H, Prevost D (2005) Ecology of plant growth promoting rhizobacteria. In: Siddiqui ZA
(ed) PGPR: biocontrol and biofertilization. Springer, Dordrecht, pp 1–38
Argueso CT, Ferreira FJ, Kieber JJ (2009) Environmental perception avenues: the interaction of
cytokinin and environmental response pathways. Plant Cell Environ 32(9):1147–1160
Aroca R, Ruiz-Lozano JM, Zamarreno AM, Paz JA, García-Mina JM, Pozo MJ, Lopez-Raez JA
(2013) Arbuscular mycorrhizal symbiosis influences strigolactone production under salinity
and alleviates salt stress in lettuce plants. J Plant Physiol 170:47–55
Babalola OO (2010) Beneficial bacteria of agricultural importance. Biotechnol Lett 32:1559–1570
Bagyalakshmi B, Ponmurugan P, Marimuthu S (2012) Influence of potassium solubilizing bacteria
on crop productivity and quality of tea (Camellia sinensis). Afr J Agric Res 7:4250–4259
Bahadur I, Meena VS, Kumar S (2014) Importance and application of potassic biofertilizer in
Indian agriculture. Int Res J Biol Sci 3:80–85
Bahadur I, Maurya BR, Kumar A, Meena VS, Raghuwanshi R (2016a) Towards the soil sustain-
ability and potassium-solubilizing microorganisms. In: Meena VS, Maurya BR, Verma JP,
Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture. Springer,
India, pp 225–266. doi:10.1007/978-81-322-2776-2_18
Bahadur I, Maurya BR, Meena VS, Saha M, Kumar A, Aeron A (2016b) Mineral release dynamics
of tricalcium phosphate and waste muscovite by mineral-solubilizing rhizobacteria isolated
from indo-gangetic plain of India. Geomicrobiol J. doi:10.1080/01490451.2016.1219431
Basak BB, Biswas DR (2009) Influence of potassium solubilizingmicroorganisms (Bacillus muci-
laginosus) and waste mica on potassium uptake dynamics by sudan grass (Sorghum vulgare
Pers.) grown under two Alfisols. Plant Soil 317(1–2):235–255
Bharti N, Barnawal D, Wasnik K, Tewari SK, Kalra A (2016) Co-inoculation of
Dietzianatronolimnaea and Glomus intraradiences with vermicompost positively influences
Ocimumbasilicum growth and resident microbial community structure in salt affected low fer-
tility soils. Appl Soil Ecol 100:211–225
Bhattacharyya P, Jha D (2012) Plant growth-promoting rhizobacteria (PGPR): emergence in agri-
culture. W J Microbiol Biotech 28:1327–1350
Bloemberg GV, Lugtenberg BJJ (2001) Molecular basis of plant growth promotion and biocontrol
by rhizobacteria. Curr Opin Plant Biol 4:343–350
Boddey RM, Urquiaga S, Reis V, Döbereiner J (1991) Biological nitrogen fixation associated with
sugar cane. Plant Soil 137:111–117
Cassán FD, Garcia de Salamone I (2008) Azospirillum sp.: Cell physiology, plant interactions and
agronomic research in Argentina. Asociación Argentina de Microbiologia, Argentina, 276 p
Chabot R, Antoun H, Cescas MP (1993) Stimulation de la croissance du maiset de la laitue romaine
par desmicroor- ganismesdissolvant le phosphoreinorganique. Can J Microbiol 39:941–947
Choudhary DK, Johri BN (2009) Interactions of Bacillus spp. and plants with special reference to
induced systemic resistance (IRS). Microbiol Res 164:493–513
Chung H, Park M, MadhaiyanaM SS, Song J, Cho H, Sa T (2005) Isolation and characterization
of phosphate solubilizing bacteria from the rhizosphere of crop plants of Korea. Soil Biol
Biochem 3:1970–1974
Compant SW, Duffy B, Nowak J, Clement C, Barka EA (2005) Use of plant growth-promoting
bacteria for biocontrol of plant diseases: principles, mechanisms of action, and future pros-
pects. Appl Environ Microb 71:4951–4959
Coyne MS (1999) Soil microbiology: an exploratory approach. Delmar publishers. pp 139–158;
279–289
Dakora FD, Phillips DA (2002) Root exudates as mediators of mineral acquisition in low-nutrient
environments. Plant Soil 245:35–47. doi:10.1023/A:1020809400075
1 Plant Beneficial Rhizospheric Microbes (PBRMs): Prospects for Increasing… 23
Dardanelli MS, Carletti SM, Paulucci NS, Medeot DB, Rodriguez Caceres EA, Vita FA, Bueno
M, Fumero MV, Garcia MB (2010) Benefits of plant growth-promoting rhizobacteria and rhi-
zobia in agriculture. In: Maheshwari DK (ed) Plant growth and health promoting bacteria,
Microbiology monographs, vol 18. Springer, Berlin, pp 1–20
Das I, Pradhan M (2016) Potassium-solubilizing microorganisms and their role in enhancing
soil fertility and health. In: Meena VS, Maurya BR, Verma JP, Meena RS (eds) Potassium
solubilizing microorganisms for sustainable agriculture. Springer, India, pp 281–291.
doi:10.1007/978-81-322-2776-2_20
Desbrosses G, Kopka C, Ott T, Udvardi MK (2004) Lotus japonicusLjKUP is induced late during
nodule development and encodes a potassium transporter of the plasma membrane. Mol Plant-
Microbe Interact 17:789–797
Dimkpa CO, Merten D, Svatos A, Büchel G, Kothe E (2009) Siderophores mediate reduced and
increased uptake of cadmium by Streptomyces tendae F4 and sunflower (Helianthus annuus),
respectively. J Appl Microbiol 5:687–1696
Dominguez-Nunez JA, Benito B, Berrocal-Lobo M, Albanesi A (2016) Mycorrhizal fungi: role
in the solubilization of potassium. In: Meena VS, Maurya BR, Verma JP, Meena RS (eds)
Potassium solubilizing microorganisms for sustainable agriculture. Springer, India, pp 77–98.
doi:10.1007/978-81-322-2776-2_6
Dotaniya ML, Meena VD, Basak BB, Meena RS (2016) Potassium uptake by crops as well
as microorganisms. In: Meena VS, Maurya BR, Verma JP, Meena RS (eds) Potassium
solubilizing microorganisms for sustainable agriculture. Springer, India, pp 267–280.
doi:10.1007/978-81-322-2776-2_19
Drake JE, Darby BA, Giasson MA, Kramer MA, Phillips RP, Finzi AC (2013) Stoichiometry con-
strains microbial response to root exudation- insights from a model and a field experiment in a
temperate forest. Biogeosciences 10:821–838
Drogue B, Sanguin H, Borland S, Prigent-Combaret C, Wisniewski-Dyé F (2014) Genome wide
profiling of Azospirillumlipoferum 4B gene expression during interaction with rice roots.
FEMS Microbiol Ecol 87:543–555
Egamberdiyeva D (2007) The effect of plant growth promoting bacteria on growth and nutrient
uptake of maize in two different soils. Appl Soil Ecol 36:184–189
Egamberdiyeva D, Islam KR (2008) Salt tolerant rhizobacteria: plant growth promoting traits and
physiological characterization within ecologically stressed environment. In: Ahmad I, Pichtel
J, Hayat S (eds) Plant-bacteria interactions: strategies and techniques to promote plant growth.
Wiley-VCH, Weinheim, pp 257–281
Farajzadeh D, Yakhchali B, Aliasgharzad N, Sokhandan-Bashir N, Farajzadeh M (2012) Plant
growth promoting characterization of indigenous Azotobacteria isolated from soils in Iran.
Curr Microbiol 64:397–403
Glick B (2012) Plant growth-promoting bacteria: Mechanisms and applications. Scientifica
2012:1–15
Glick BR, Todorovic B, Czarny J, Cheng Z, Duan J, McConkey B (2007) Promotion of plant
growth by bacterial ACC deaminase. Crit Rev Plant Sci 26:227–242
Govindarajan M, Balandreau J, Kwon SW, Weon HY, Lakshminarasimhan C (2008) Effects of the
inoculation of Burkholderiavietnamensis and related endophytic diazotrophic bacteria on grain
yield of rice. Microb Ecol 55:21–37
Haas D, Defago G (2005) Biological control of soil-borne pathogens by fluorescent pseudomo-
nads. Nat Rev Microbiol 3:307–319
Haichar F, Marol C, Berge O, Rangel-Castro JI, Prosser JI, Balesdent J, Heulin T, Achouak W
(2008) Plant host habitat and root exudates shape soil bacterial community structure. ISME
J:1221–1230
Harman WL, Gerik TJ, Williams JR (2004) CROPMAN: a crop production/environmental simu-
lation tool to enhance agricultural sustainability. In: Dilawari VK, Brar LS, Jalota SK (eds)
Proceedings of sustaining agriculture: problems and prospects. Punjab Agricultural University,
Ludhiana., November 9–11, 2004, pp 22–30
24 A. Sarkar et al.
Hartmann A, Schmid M, van Tuinen D, Berg G (2009) Plant-driven selection of microbes. Plant
Soil 321:235–257
Hider RC, Kong X (2010) Chemistry and biology of siderophores. Nat Prod Rep 27(5):637
Hodge A (2017) Accessibility of inorganic and organic nutrients for mycorrhizas. Mycorrhizal
mediation of soil fertility, structure and carbon storage Chapter-8 pp. 129–148
Hussain N, Mujeeb F, Tahir M, Khan GD, Hassan NM, Bari A (2002) Effectiveness of Rhizobium
under salinity stress. Asian J Plant Sci 1:12–14
Hutchens SE, Valsami JE, Eldowney MS (2003) The role of heterotrophic bacteria in feldspar dis-
solution. Min Mag 67:1151–1170
Ikeda AG, Bassani LL, Adamoski D, Stringari D, Cordeiro VK, Glienke C, Maria Steffens BR,
Hungria M, Galli-Terasawa LV (2013) Morphological and genetic character- ization of endo-
phytic bacteria isolated from roots of differ- ent maize genotypes. MicrobEcol 65:154–160
Jacobsen BJ, Zidack NK, Larson BJ (2004) The role of -based biological control agents in inte-
grated pest management systems: plant diseases. Phytopathology 94(11):1272–1275
Jaiswal DK, Verma JP, Prakash S, Meena VS, Meena RS (2016) Potassium as an important plant
nutrient in sustainable agriculture: a state of the art. In: Meena VS, Maurya BR, Verma JP,
Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture. Springer,
India, pp 21–29. doi:10.1007/978-81-322-2776-2_2
Jat LK, Singh YV, Meena SK, Meena SK, Parihar M, Jatav HS, Meena RK, Meena VS (2015)
Does integrated nutrient management enhance agricultural productivity? J Pure Appl Microbiol
9(2):1211–1221
Jha Y, Subramanian RB (2016) Regulation of plant physiology and antioxidant enzymes for
alleviating salinity stress by potassium-mobilizing bacteria. In: Meena VS, Maurya BR,
Verma JP, Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture.
Springer, India, pp 149–162. doi:10.1007/978-81-322-2776-2_11
Katznelson H, Lochhead AG, Timonin MI (1948) Soil microorganisms and the rhizosphere. Bot
Rev 14:543–587
Khan MS, Zaidi A, Wani PA (2009) Role of phosphate- solubilizing microorganisms in sustainable
agriculture – a review. Agron Sustain Dev 27:29–43
Kim H, Park J, Choi SW, Choi KH, Lee G, Ban S, Lee C, Kim CS (2003) Isolation and character-
ization of Bacillus strains for biological control. J Microbiol 41(3):196–201
Kim YC, Leveau J, Gardener BBM, PiersonEA PLS, Ryu CM (2011) The multifactorial basis for
plant health promotion by plant-associated bacteria. Appl Environ Microbiol 77:1548–1555
Kobayashi T, Nishizawa NK (2012) Iron uptake, translocation and regulation in higher plants.
Annu Rev Plant Bio 63:11–152
Krewulak HD, Vogel HJ (2008) Structural biology of bacterial iron uptake. Biochim Biophys Acta
1778:1781–1804
Kumar VK, Raju SK, Reddy MS, Kloepper JW, Lawrence KS, Groth DE, Miller ME, Sudini H,
Binghai D (2009) Evaluation of commercially available PGPR for control of rice sheath blight
caused by Rhizoctoniasolani. J Pure Appl Microbiol 3(2):485–488
Kumar A, Bahadur I, Maurya BR, Raghuwanshi R, Meena VS, Singh DK, Dixit J (2015) Does
a plant growth-promoting rhizobacteria enhance agricultural sustainability? J Pure Appl
Microbiol 9:715–724
Kumar A, Meena R, Meena VS, Bisht JK, Pattanayak A (2016a) Towards the stress management
and environmental sustainability. J Clean Prod 137:821–822
Kumar A, Patel JS, Bahadur I, Meena VS (2016b) The molecular mechanisms of KSMs for
enhancement of crop production under organic farming. In: Meena VS, Maurya BR, Verma JP,
Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture. Springer,
India, pp 61–75. doi:10.1007/978-81-322-2776-2_5
Kumar A, Maurya BR, Raghuwanshi R, Meena VS, Islam MT (2017) Co-inoculation with
Enterobacter and Rhizobacteria on yield and nutrient uptake by wheat (Triticum aestivum L.)
in the alluvial soil under indo-gangetic plain of India. J Plant Growth Regul. doi:10.1007/
s00344-016-9663-5
1 Plant Beneficial Rhizospheric Microbes (PBRMs): Prospects for Increasing… 25
Meena RS, Meena VS, Meena SK, Verma JP (2015a) The needs of healthy soils for a healthy
world. J Clean Prod 102:560–561
Meena RS, Meena VS, Meena SK, Verma JP (2015b) Towards the plant stress mitigate the agricul-
tural productivity: a book review. J Clean Prod 102:552–553
Meena VS, Maurya BR, Meena RS (2015c) Residual impact of wellgrow formulation and NPK on
growth and yield of wheat (Triticum aestivum L.) Bangladesh J Bot 44(1):143–146
Meena VS, Maurya BR, Verma JP, Aeron A, Kumar A, Kim K, Bajpai VK (2015d) Potassium
solubilizing rhizobacteria (KSR): isolation, identification, and K-release dynamics from waste
mica. Ecol Eng 81:340–347
Meena VS, Meena SK, Verma JP, Meena RS, Ghosh BN (2015e) The needs of nutrient use efficiency
for sustainable agriculture. J Clean Prod 102:562–563. doi:10.1016/j.jclepro.2015.04.044
Meena VS, Verma JP, Meena SK (2015f) Towards the current scenario of nutrient use efficiency in
crop species. J Clean Prod 102:556–557. doi:10.1016/j.jclepro.2015.04.030
Meena RK, Singh RK, Singh NP, Meena SK, Meena VS (2016a) Isolation of low temperature sur-
viving plant growth-promoting rhizobacteria (PGPR) from pea (Pisum sativum L.) and docu-
mentation of their plant growth promoting traits. Biocatalysis and Agricultural. Biotechnology
4:806–811
Meena RS, Bohra JS, Singh SP, Meena VS, Verma JP, Verma SK, Sihag SK (2016b) Towards the
prime response of manure to enhance nutrient use efficiency and soil sustainability a current
need: a book review. J Clean Prod 112(1):1258–1260
Meena SK, Rakshit A, Meena VS (2016c) Effect of seed bio-priming and N doses under varied
soil type on nitrogen use efficiency (NUE) of wheat (Triticum aestivum L.) under greenhouse
conditions. Biocatal Agric Biotechnol 6:68–75
Meena VS, Bahadur I, Maurya BR, Kumar A, Meena RK, Meena SK, Verma JP (2016d) Potassium-
solubilizing microorganism in evergreen agriculture: an overview. In: Meena VS, Maurya BR,
Verma JP, Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture.
Springer, India, pp 1–20. doi:10.1007/978-81-322-2776-2_1
Meena VS, Meena SK, Bisht JK, Pattanayak A (2016e) Conservation agricultural practices in
sustainable food production. J Clean Prod 137:690–691
Mendes R, Garbeva P, Raaijmakers JM (2013) The rhizosphere microbiome: significance of plant
beneficial plant pathogenic and human pathogenic microorganisms. FEMS Microbiol Rev
37:634–663
Montano FP, Villegas CA, Bellogin RA, del Cerro P, Espuny MR, Guerrero IJ, FJL B, Ollero FJ,
Cubo T (2014) Plant growth promotion in cereal and leguminous agricultural important plants:
from microorganism capacities to crop production. Microbiol Res 169:325–336
Morgan JA, Lecain DR, Pendall E, Blumenthal DM, Kimball BA, Carrillo Y et al (2011) C4 grasses
prosper as carbon dioxide eliminates desiccation in warmed semi-arid grassland. Nature
476:202–205. doi:10.1038/nature10274
Morris SJ, Blackwood CB (2015) The ecology of soil biota and their function. Soil Microbiology,
Ecology and Biodiversity (Fourth Edition) Chapter-10 pp. 273–309
Nishanth D, Biswas DR (2008) Kinetics of phosphorus and potassium release from rockphos-
phate and waste mica enriched compost and their effect on yield and nutrient uptake by wheat
(Triticumaestivum). Bioresour Technol 99:3342–3354
Parewa HP, Yadav J, Rakshit A, Meena VS, Karthikeyan N (2014) Plant growth promoting rhizo-
bacteria enhance growth and nutrient uptake of crops. Agric Sustain Dev 2(2):101–116
Prakash S, Verma JP (2016) Global perspective of potash for fertilizer production. In: Meena VS,
Maurya BR, Verma JP, Meena RS (eds) Potassium solubilizing microorganisms for sustainable
agriculture. Springer, India, pp 327–331. doi:10.1007/978-81-322-2776-2_23
Priyadharsini P, Muthukumar T (2016) Interactions between arbuscular mycorrhizal fungi and
potassium-solubilizing microorganisms on agricultural productivity. In: Meena VS, Maurya
BR, Verma JP, Meena RS (eds) Potassium solubilizing microorganisms for sustainable agricul-
ture. Springer, India, pp 111–125. doi:10.1007/978-81-322-2776-2_8
Raghavendra MP, Nayaka NC, Nuthan BR (2016) Role of rhizosphere microflora in potas-
sium solubilization. In: Meena VS, Maurya BR, Verma JP, Meena RS (eds) Potassium
1 Plant Beneficial Rhizospheric Microbes (PBRMs): Prospects for Increasing… 27
Singh BK, Millard P, Whiteley AS, Murrell JC (2004) Unravelling rhizosphere-microbial inter-
actions: opportunitiesand limitations. Trends Microbiol 12:386–393. PMID:15276615; http://
dx.doi. org/10.1016/j.tim.2004.06.008
Singh NP, Singh RK, Meena VS, Meena RK (2015) Can we use maize (Zea mays) rhizobacteria as
plant growth promoter? Vegetos 28(1):86–99. doi:10.5958/2229-4473.2015.00012.9
Singh M, Dotaniya ML, Mishra A, Dotaniya CK, Regar KL, Lata M (2016) Role of biofertil-
izers in conservation agriculture. In: Bisht JK, Meena VS, Mishra PK, Pattanayak A (eds)
Conservation agriculture: an approach to combat climate change in Indian Himalaya. Springer,
Singapore, pp 113–134. doi:10.1007/978-981-10-2558-7_4
Sommers E, Vanderleyden J, Srinivasan M (2004) Rhizosphere bacterial signalling: a love parade
beneath our feet. Crit Rev Microbiol 30:205–240
Steenhoudt O, Vanderleyden J (2000) Azospirillum, a free-living nitrogen-fixing bacterium closely
associated with the grasses: genetic, biochemical and ecological aspect. FEMS Microbiol Rev
24(4):487–506
Takagi S, Nomoto K, Takemoto T (1984) Physiological aspect of mugineic acid, a possible phyto-
siderophore of graminaceous plants. J Plant Nutri 7:469–477
Teotia P, Kumar V, Kumar M, Shrivastava N, Varma A (2016) Rhizosphere microbes: potassium
solubilization and crop productivity-present and future aspects. In: Meena VS, Maurya BR,
Verma JP, Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture.
Springer, India, pp 315–325. doi:10.1007/978-81-322-2776-2_22
Velazquez E, Silva LR, Ramírez-Bahena MH, Peix A (2016) Diversity of potassium-solubilizing
microorganisms and their interactions with plants. In: Meena VS, Maurya BR, Verma JP,
Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture. Springer,
India, pp 99–110. doi:10.1007/978-81-322-2776-2_7
Verma R, Maurya BR, Meena VS (2014) Integrated effect of bio-organics with chemical fertilizer
on growth, yield and quality of cabbage (Brassica oleracea var capitata). Indian J Agric Sci
84(8):914–919
Verma P, Yadav AN, Khannam KS, Panjiar N, Kumar S, Saxena AK, Suman A (2015a) Assessment
of genetic diversity and plant growth promoting attributes of psychrotolerant bacteria allied with
wheat (Triticumaestivum) from the northern hills zone of India. Ann Microbiol. doi:10.1007/
s13213-014-1027-4
Verma JP, Jaiswa DK, Meena VS, Meena RS (2015b) Current need of organic farming for enhanc-
ing sustainable agriculture. J Clean Prod 102:545–547
Verma JP, Jaiswal DK, Meena VS, Kumar A, Meena RS (2015c) Issues and challenges about sus-
tainable agriculture production for management of natural resources to sustain soil fertility and
health. J Clean Prod 107:793–794
van der Wal A, de Boer W (2017) Dinner in the dark: illuminating drivers of soil organic matter
decomposition. Soil Biol Biochem 105:45–48
Vleesschauwer D (2007) Using Serratia plymuthica to control fungal pathogens of plants. CAB
Reviews 2(046)
Wani SA, Chand S, Ali T (2013) Potential use of Azotobacter chroococcum in crop production: an
overview. Curr Agric Res J 1:35–38
Whipps J (2001) Microbial interactions and biocontrol in the rhizosphere. J Exp Bot 52:487–511
Wiren N, Khodr H, Hider RC (2000) Hydroxylated phytosiderophore species possess and enhanced
chelate stability and affinity for iron(III). Plant Physiol 124:1149–1157
Yadav BK, Sidhu AS (2016) Dynamics of potassium and their bioavailability for plant nutrition.
In: Meena VS, Maurya BR, Verma JP, Meena RS (eds) Potassium solubilizing microorganisms
for sustainable agriculture. Springer, India, pp 187–201. doi:10.1007/978-81-322-2776-2_14
Yang J, Kloepper JW, Ryu CM (2009) Rhizosphere bacteria help plants tolerate abiotic stress.
Trends Plant Sci 14(1):1–4
Yasin M, Munir I, Faisal M (2016) Can Bacillus spp. enhance K+ uptake in crop species. In:
Meena VS, Maurya BR, Verma JP, Meena RS (eds) Potassium solubilizing microorganisms
for sustainable agriculture. Springer, India, pp 163–170. doi:10.1007/978-81-322-2776-2_12
1 Plant Beneficial Rhizospheric Microbes (PBRMs): Prospects for Increasing… 29
Abstract
The importance of soils and its function is conclusive; the soil is a living entity
that needs to be improved under sustainable approach. It is apparent that soil
biota contributes substantially to effective soil functioning and maintenance of
soil sustainability. The increase of food production to meet out the food security
demand of an ever-increasing population has been highly dependent on inte-
grated nutrient management input but ignores the potential of bioagents. The
significance of rhizosphere microorganism can be enormous as they are able in
managing nutrient transformation, nutrient acquisition and use, and thus crop
sustainability. The soil sustainability and nutrient use efficiency (NUE) can be
enhanced by modification of rhizosphere through the best management
approaches such as agronomic practices, nutrient management, use of organic
and biofertilizers, and regulation of rhizosphere processes.
Keywords
Rhizosphere • Soil sustainability • Nutrient availability • PGPR • Bioremediation
2.1 Introduction
Agriculture in the twenty-first century faces more challenges, like decline in pro-
ductivity and degradation in soil quality. As per the United Nations estimates, the
global human population is projected to reach ~9 billion by 2050 (Wood 2001). The
feeding of our ever-increasing population is one of the largest challenges for main-
taining the worldwide agricultural sustainability. The soil being a critical compo-
nent on the earth is not only for sufficient food production but also for maintaining
worldwide environmental sustainability for the next generation (Meena et al. 2013a;
Bahadur et al. 2014; Maurya et al. 2014; Kumar et al. 2015, 2017a; Ahmad et al.
2016; Zahedi 2016). These existing technologies have reached to its plateau and are
hitherto extremely difficult to further increase the food production. Therefore,
exploring nonconventional resources is an urgent need not only to increase the
demand of ever-increasing population but also to sustain our ecosystem from the
further degradation. Sustainability in the agriculture production cannot be attained
without microbiological population in soil under the present circumstances
(Vaxevanidou et al. 2015; Patil et al. 2014).
These efficient microorganisms are key factor of the living biomass of the soil
system (Whitman et al. 1998). Soil microorganisms play significant roles in agricul-
ture mainly by improving plant nutrition and soil health, as well as soil quality
(Barea 2004; Barea et al. 2013; Lugtenberg 2015) and degrading organic pollutants.
The function of agriculture ecosystem depends largely on soil microbial dynamics
(Barea 2004). These efficient soil microorganisms maintain the organic matter
(OM) mineralization and thus regulate nutrient cycling. The beneficial saprophytic
rhizosphere microbes improve plant performance work as a decomposer of OM
substances, plant growth-promoting bacteria/rhizobacteria (PGPB/PGPR), and
other plant growth-promoting (PGP) activities. The efficient PGPR must have the
ability to survive and multiply in rhizosphere microbes with indigenous microbes,
at least for the time needed to express their beneficial PGP (Martinez-Viveros et al.
2010). The rhizosphere microorganisms improve nutrient cycling processes that
include N fixation, P mobilization, and the release of other nutrients to soil solution
for plant nutrition (Richardson et al. 2009; Barea and Richardson 2015).
The rhizosphere soils contain efficient microbes, and researcher differentiates
them into beneficial and harmful categories based on their beneficial effect on soil
sustainability, crop growth, and yield (Welbaum et al. 2004; Bais et al. 2006;
Brimecombe et al. 2007; Jat et al. 2015). Microbes play vital roles in soil sustain-
ability and nutrient acquisition. Without soil microbes life on the planet would not
be possible. An increased knowledge of rhizospheric chemistry-based symbioses in
plants could provide potential ways of developing sustainable agriculture with mini-
mal risk to the environment. Therefore, rhizosphere microorganisms have a key role
to improve NUE and crop sustainability simultaneously (Bahadur et al. 2016a, b;
Das and Pradhan 2016; Dominguez-Nunez et al. 2016; Verma et al. 2014, 2015b;
Yadav and Sidhu 2016; Yasin et al. 2016).
2 Rhizosphere Microorganisms Towards Soil Sustainability and Nutrient Acquisition 33
Providing as well as feeding rapidly growing human population is one of the great-
est challenges worldwide. The requirement of food will be ~29 Mt, while their
availability will be only ~22 Mt by 2020. Best management practices (BMPs) and
recommended management practices (RMPs) of soil system ensure that mineral
nutrients do not become deficient or toxic to plants as well as humans and that
appropriate crop/mineral nutrients enter the food chain. Soil management is impor-
tant, both directly and indirectly, to crop productivity, environmental sustainability,
and human health. Because of the projected increase in worldwide population and
the consequent necessity for the nutritional security, the management of soils will
become increasingly important in the coming years because the traditional agricul-
ture will not be able to meet the needs of the next generation (Altieri 2004).
The rhizosphere is the narrow region of soil system that is immediately near to the
root surface and that is more affected by rhizospheric phenomena. The term “rhizo-
sphere” was first time used by Lorenz Hiltner in 1904. The root exudates include
carbohydrates, sugar, organic acids, vitamins, flavonoids, nucleotides, enzymes,
hormones, and volatile compounds, inorganic ions, and gaseous molecules. The
exudates act as messengers that stimulate interactions between roots and soil organ-
isms. Therefore, rhizosphere is the most dynamic environment in the soil with a
high microbial diversity. It is well established that microorganism’s population is
higher in rhizosphere (Table 2.1) than bulk soil (Brimecombe et al. 2001; Bahadur
et al. 2017; Verma et al. 2017b; Kumar et al. 2017a).
In the rhizosphere, the populations of microorganisms may fluctuate from few thou-
sands to millions (Nihorimbere et al. 2011). This plant-microbe interaction has shown
many beneficial effects on plant growth and developments (Singh et al. 2004), PGP
increased availability and uptakes of nutrients, and disease suppression (Morrissey
et al. 2004; Haas and Defago 2005; Mendes et al. 2011). However, increased resistance
to abiotic and biotic stresses (Zolla et al. 2013; Badri et al. 2013), all these leads to
increases in crop productivity and soil sustainability as well as nutrient availability
(Bardgett et al. 2014; Nath et al. 2017; Sarkar et al. 2017; Verma et al. 2017a).
Table 2.1 Presence of microbes in the rhizosphere and non-rhizosphere soil system
Rhizosphere Non-rhizosphere
Microorganisms Microbes g−1 dry soil R:S ratio
Bacteria 1200 × 106 53 × 106 23.0
Fungi 12 × 105 1 × 105 12.0
Actinomycetes 46 × 106 7 × 106 7.0
Algae 5 × 103 27 × 103 0.2
34 M. Prasad et al.
The OM has been considered as one of the most important components in improv-
ing soil health and agricultural sustainability. The rhizosphere microorganisms play
an important role in regulating soil organic matter (SOM) decomposition and nutri-
ent cycling due to the direct influences by roots that are often referred as rhizosphere
processes. These rhizosphere processes may include exudates, water uptake, nutri-
ent mobilization by roots and microorganisms, and SOM decomposition. These effi-
cient rhizosphere processes are major paths for nutrient and water availability.
Therefore, rhizosphere effect on the SOM decomposition is significant in mediating
plant-soil interactions.
SOM decomposition is mediated by rhizosphere processes (Sallih and Bottner
1988; Cheng and Coleman 1990). This OM in the soil system is more or less con-
tinuously transformed by different chemical and biochemical mechanisms into vari-
ous ranges of compounds (Marschner and Rengel 2007). The SOM contains various
compounds which are the main energy source for the soil biota. Soil organisms have
2 Rhizosphere Microorganisms Towards Soil Sustainability and Nutrient Acquisition 35
appropriate biochemistries and life processes to embezzle such energy for their
growth; in doing so, the compounds are further transformed and cycled between
compartments (Masood and Bano 2016; Meena et al. 2016a; Shrivastava et al. 2016;
Sindhu et al. 2016; Singh et al. 2016; Kumar et al. 2017b).
Table 2.3 Microbiological properties in bulk and rhizosphere soils of different plantations
MBC MBN MBP
Tree species mg kg−1 Microbial C/N
Siberian elm Bulk soil 36.8(4.9) 7.0(0.6) 1.8(0.1) 5.2(0.8)
Rhizosphere soil 52.7(5.2) 9.4(1.0) 2.1(0.2) 5.6(1.0)
T-test ** * NS NS
Simon poplar Bulk soil 73.6(7.2) 6.7(0.2) 2.6(0.1) 10.9(1.2)
Rhizosphere soil 90.7(6.4) 8.2(0.6) 2.8(0.2) 11.1(1.1)
T-test * * NS NS
Mongolian pine Bulk soil 45.6(3.2) 4.9(0.5) 1.6(0.1) 9.4(1.0)
Rhizosphere soil 48.4(4.2) 8.0(0.9) 1.8(0.2) 6.1(0.7)
T-test NS ** NS *
Adopted from Zhao et al. (2010)
Data are means with standard errors given in parentheses (n = 5). NS denotes no significant differ-
ence (by paired samples T-test)
* Significant differences between bulk and rhizosphere soils at P < 0.05
** Significant differences between bulk and rhizosphere soils at P < 0.01
The bacteria present in the rhizosphere act as PGPR; they have the ability to enhance
PGP atmospheric N fixation, hormone production, specific enzymatic activity, and
plant and insect protection by producing antibiotic and other pathogen-depressing
substances (Kamnev and Lelie 2000). However, the N2-fixing, PSB, and KSB are
important for crop plants as they increase N, P, and K uptake and play a crucial role
as PGPR in the biofertilization (Zhang et al. 2013). In general, huge bulk inorganic/
chemical fertilization is applied to replenish soil N and P demand with the resultant
in high cost and environmental risk. Most of the P-fixed in the form of insoluble
compounds is unavailable to plants. Thus, the application of such efficient microbes
as eco-friendly biological agents may contribute to minimize the use of expensive P
and K fertilizers.
A broad range of efficient microorganisms carry out biocontrol activities in the rhi-
zosphere soils. These microbes play a major role to control diseases, which is a
form of biological control and is an environment-friendly approach. Many rhizo-
sphere bacteria have been reported to produce antifungal compounds like HCN,
phenazines, pyrrolnitrin, 2,4-diacetylphloroglucinol, and pyoluteorin (Bhattacharyya
and Jha 2012). The biocontrol ability of the PGPR depends on a wide variety of
traits, such as the production of various antibiotic compounds, iron chelators, and
exoenzymes (Meena et al. 2014b, 2015c, d; Prakash and Verma 2016).
2 Rhizosphere Microorganisms Towards Soil Sustainability and Nutrient Acquisition 37
Table 2.4 Effect of PGPR on various PGP activities under sustainable agriculture
PGPRs Responses Crop References
Pseudomonas and Stimulates growth and Chickpea Rokhzadi et al. (2008)
Azotobacter yield
P. putida strain R-168 Enhance seed Maize Gholami et al. (2009)
germination, growth
parameters
P. fluorescens strain R-93, Increase growth, leaf Banana Kavino et al. (2010)
P. fluorescens DSM 50090, nutrient contents and
P. putida DSM291, and yield
A. lipoferum DSM 1691
P. putida strains R-168 and Improves seed Maize Nezarat and Gholami
DSM-291 germination, seedling (2009)
growth and yield
R. leguminosarum and Enhance yield and P Wheat Afzal and Asghari (2008)
Pseudomonas sp. uptake
P. putida strain Growth stimulation Tomato Gravel et al. (2007)
Azospirillum brasilense Improve growth Wheat and Dobbelaere et al. (2002)
attributes maize
P. fluorescens strain Growth and yield Pearl millet Niranjan et al. (2003)
Azotobacter and Growth and productivity Canola Yasari and Patwardhan
Azospirillum strains (2007)
P. alcaligenes PsA15 and Enhance nutrient uptake Maize Egamberdiyeva (2007)
B. polymyxa
Rhizobium leguminosarum Direct growth promotion Canola and Noel et al. (1996)
lettuce
The soil is an important habitat for thousands of microbes (Table 2.4); these effi-
cient microbes are habitats between soil particles and soil organic matter in rhizo-
sphere (Akhtar et al. 2013). Many soil microbes are capable of degrading toxic
compounds including herbicides, pesticides, solvents, and other organic compounds
and might provide a reasonable and effective measure of disposing toxic compounds
(Murali and Mehar 2014).
Due to the degrading and sequestration capability of the microbial communities
toward the toxic compounds and heavy metals (HMs), microbes have been used for
bioremediation of soil contaminated with HMs (Table 2.5) (Dell’Amico et al. 2005;
Kao et al. 2006; Umrania 2006).
The organic pollutants can be degraded by plants/crops/microbes through bio-
chemical reactions taking place within the rhizosphere. The phytodegradation of
organic pollutants may also be enhanced by rhizospheric activity (Mirsal 2004). The
AMF were also demonstrated to PGP and degradation of toxic pollutants in soils
with increased pollutant concentrations especially ectomycorrhizal fungi (Table 2.6).
They promote the establishment of plants at HM-contaminated soil by immobiliz-
ing heavy metals in the soil, thereby reducing the availability of metals to plants
(Jentschke and Godbold 2000; Meharg and Cairney 2000; Fomina et al. 2005).
38 M. Prasad et al.
Diverse environmental stresses like drought, salinity, and nutrient deficiencies are
connected to soil-derived limitations. These stresses are inducing a series of mor-
phological, physiological, biochemical, and molecular changes in the plant root sys-
tem to adapt to the stress conditions and/or to modify the environment from which
the stress is originated. Variation in the soil nutrient composition is a common con-
dition in agricultural systems. Indeed, only ~30 to 50% of applied N fertilizers and
~10 to 20% of P fertilizers are used in crops (Adesemoye and Kloepper 2009).
Furthermore, there is a general trend worldwide in reducing the use of chemical
inputs in agriculture due to environmental concerns (Meena et al. 2013c, 2016e;
Parewa et al. 2014). Water and nutrient acquisition from soils is governed by plant
roots and their interaction with the abiotic and biotic components of soil (Hartmann
2 Rhizosphere Microorganisms Towards Soil Sustainability and Nutrient Acquisition 39
et al. 2008). The soil biotic component includes microorganisms that are living
symbiotically or in association with plant roots. These microorganisms might
enhance acquisition of nutrients in low-nutrient soils and alleviate the effects of low
water availability. But there are only few studies that provided information regard-
ing rhizosphere soil microorganisms to plants in mitigating with stresses derived
from nutrient and water availability (Saleem et al. 2007; Hayat et al. 2010; Kang
et al. 2010; Smith et al. 2010; Meena et al. 2015e).
It is well known that nutrient has unsounded effects on many aspects of root
development, thus accompanying rhizosphere process (Zhang et al. 2010; Meena
et al. 2015f). Most of the mineral nutrients were taken up by plants through the
rhizosphere where root exudates play a dominant role in driving interactions among
soil-plant-microbe system. However, the plant root exudates consist of a complex
mixture of various acids, sugars, and enzymes, which have major direct or indirect
effects on the acquisition of mineral nutrients required for PGP activities (Dakora
and Phillips 2002). The rhizospheric chemistry/phenomena can enhance P mobili-
zation and acquisition from soil by plants (Zhang et al. 2010). It is suggested that
optimizing crop combination and BMPs, RMPs, and INM in intercropping systems
through better understanding of rhizosphere interactions provides an effective
approach to improve sustainable crop production with high yield, significantly
higher NUE (Zhang et al. 2012).
40 M. Prasad et al.
Rhizospheric modification and root distribution along with the cropping system
play a significant role toward the sustainable rhizosphere management. The subsur-
face irrigation systems can be adopted to improve soil characteristics and root dis-
tribution/modification, in relation to the physical properties of the soil and
fertilization practices (Clothier and Green 1994; Yang et al. 2004).
The organic farming (OF) system has become popular due to consumer concern
with human health and the environment safety. Organic fertilization for the nutrient
supply is more balanced, which helps to maintain soil sustainability and crop pro-
ductivity. Among the beneficial effects of organic manures include (1) enhancement
of soil biological activity, (2) enhancement of colonization of mycorrhizae, (3)
improved root growth due to better soil structure, and (4) slow release of nutrients
which contribute to the residual pool of organic N and P in the soil, reducing N
leaching loss and P fixation (Singh and Mandal 2000).
References
Adesemoye AO, Kloepper JW (2009) Plant-microbes interactions in enhanced fertilizer use effi-
ciency. Appl Microbiol Biotechnol 85:1–12
Adesemoye AO, Torbert HA, Kloepper JW (2008) Enhanced plant nutrient use efficiency with
PGPR and AMF in an integrated nutrient management system. Can J Microbiol 54:876–886
Afzal A, Asghari B (2008) Rhizobium and phosphate solubilizing bacteria improve the yield and
phosphorus uptake in wheat. Int J Agric Biol 10:85–88
Ahmad M, Nadeem SM, Naveed M, Zahir ZA (2016) Potassium-solubilizing bacteria and their
application in agriculture. In: Meena VS, Maurya BR, Verma JP, Meena RS (eds) Potassium
solubilizing microorganisms for sustainable agriculture. Springer, India, pp 293–313.
doi:10.1007/978-81-322-2776-2_21
Akhtar MS, Panwar J (2011) Arbuscular mycorrhizal fungi and opportunistic fungi: efficient root
symbionts for the management of plant parasitic nematodes. Adv Sci Eng Med 3:165–175
Akhtar MS, Siddiqui ZA (2008) Arbuscular mycorrhizal fungi as potential biprotectants against
plant pathogens. In: Siddiqui ZA, Akhtar MS, Futai K (eds) Mycorrhizae: sustainable agricul-
ture and forestry. Springer, Dordrecht, pp 61–98
Akhtar MS, Siddiqui ZA (2010) Role of plant growth promoting rhizobacteria in biocontrol of
plant diseases and sustainable agriculture. In: Maheshwari DK (ed) Plant growth and health
promoting bacteria, Microbiology monographs 18. Springer, Berlin, pp 157–196
Akhtar MS, Chali B, Azam T (2013) Bioremediation of arsenic and lead by plants and microbes
from contaminated soil. Res Plant Sci 1:68–73
Altieri MA (2004) Linking ecologists and traditional farmers in the search for sustainable agricul-
ture. Front Ecol Environ 2:35–42
Badri DV, Chaparro JM, Zhang R, Shen Q, Vivanco JM (2013) Application of natural blends
of phytochemicals derived from the root exudates of Arabidopsis to the soil reveal that
phenolic-related compounds predominantly modulate the soil microbiome. J Biol Chem
288(7):4502–4512
Bahadur I, Meena VS, Kumar S (2014) Importance and application of potassic biofertilizer in
Indian agriculture. Int Res J Biol Sci 3:80–85
Bahadur I, Maurya BR, Kumar A, Meena VS, Raghuwanshi R (2016a) Towards the soil sustain-
ability and potassium-solubilizing microorganisms. In: Meena VS, Maurya BR, Verma JP,
Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture. Springer,
India, pp 225–266. doi:10.1007/978-81-322-2776-2_18
Bahadur I, Maurya BR, Meena VS, Saha M, Kumar A, Aeron A (2016b) Mineral release dynamics
of tricalcium phosphate and waste muscovite by mineral-solubilizing rhizobacteria isolated
from indo-gangetic plain of India. Geomicrobiol J. doi:10.1080/01490451.2016.1219431
Bahadur I, Maurya BR, Meena VS, Saha M, Kumar A, Aeron A (2017) Mineral release dynamics
of tricalcium phosphate and waste muscovite by mineral-solubilizing rhizobacteria isolated
from indo-gangetic plain of India. Geomicrobiol J. doi:10.1080/01490451.2016.1219431
42 M. Prasad et al.
Bais HP, Weir TL, Perry LG, Gilroy S, Vivanco JM (2006) The role of root exudates in rhizosphere
interactions with plants and other organisms. Annu Rev Plant Biol 57(1):233–266
Bakker MG, Manter DK, Sheflin AM, Weir TL, Vivanco JM (2012) Harnessing the rhizosphere
microbiome through plant breeding and agricultural management. Plant Soil 360:1–13
Bardgett RD, Mommer L, De Vries FT (2014) Going underground: root traits as drivers of ecosys-
tem processes. Trends Ecol Evol 29:692–699
Barea JM (2004) Impacto de las micorrizas en la calidad del suelo y la productividad vegetal en
sistemas agrícolas y espacios naturales. In: Monzón de Asconegui MA, García de Salamone
IE, Miyazaki SS (eds) Biología del suelo. Transformaciones de la materia orgánica, usos y
biodiversidad de los organismos edáficos. Editorial FAUBA, Universidad de Buenos Aires,
Argentina, pp 7–11
Barea JM, Richardson AE (2015) Phosphate mobilisation by soil microorganisms. In: Lugtenberg
B (ed) Principles of plant-microbe interactions. Springer International Publishing Switzerland,
Heidelberg, pp 225–234
Barea JM, Pozo MJ, Azcón R, AzcónAguilar C (2013) Microbial interactions in the rhizosphere.
In: de Bruijn FJ (ed) Molecular microbial ecology of the rhizosphere, vol 1. Wiley Blackwell,
Hoboken, pp 29–44
Bhattacharyya PN, Jha DK (2012) Plant growth-promoting rhizobacteria (PGPR): emergence in
agriculture. World J Microbiol Biotechnol 28:1327–1350
Bianciotto V, Bonfante P (2002) Arbuscular mycorrhizal fungi: a specialized niche for rhizospheric
and endocellular bacteria. Antonie Van Leeuwenhoek 81:365–371
Bisht S, Pandey P, Aggarwal H et al (2014) Utilization of endophytic strain Bacillus sp. SBER3
for biodegradation of polyaromatic hydrocarbons (PAH) in soil model system. Eur J Soil Biol
60:67–76
Brazil GM, Kenefick L, Callanan M et al (1995) Construction of a rhizosphere Pseudomonad with
potential to degrade polychlorinated biphenyls and detection of bph gene expression in the
rhizosphere. Appl Environ Microbiol 61:1946–1952
Brimecombe MJ, De Lelj FA, Lynch JM (2001) The rhizosphere. The effect of root exudates
on rhizosphere microbial populations. In: Pinton R, Varanini Z, Nannipieri P (eds) The
rhizosphere. Biochemistry and organic substances at the soil-plant Interface. Marcel Dekker,
New York, pp 95–140
Brimecombe MJ, De Leij FAAM, Lynch JM (2007) Rhizodeposition and microbial populations.
In: Pinton R, Veranini Z, Nannipieri P (eds) The rhizosphere biochemistry and organic sub-
stances at the soil-plant interface. Taylor & Francis Group, New York
Cheng W, Coleman DC (1990) Effect of living roots on soil organic matter decomposition. Soil
Biol Biochem 22:781–787
Clark RB, Zeto SK (2000) Mineral acquisition by arbuscular mycorrhizal plants. J Plant Nutr
23:867–902
Clothier BE, Green SR (1994) Rootzone processes and the efficient use of irrigation water. Agric
Water Manag 25:1–12
Dakora FD, Phillips DA (2002) Root exudates as mediators of mineral acquisition in low nutrient
environments. Plant Soil 245:35–47
Das I, Pradhan M (2016) Potassium-solubilizing microorganisms and their role in enhancing
soil fertility and health. In: Meena VS, Maurya BR, Verma JP, Meena RS (eds) Potassium
solubilizing microorganisms for sustainable agriculture. Springer, India, pp 281–291.
doi:10.1007/978-81-322-2776-2_20
Dell’Amico E, Cavalca L, Andreoni V (2005) Analysis of rhizobacterial communities in perennial
Graminaceae from polluted water meadow soil, and screening of metal-resistant, potentially
plant growth-promoting bacteria. FEMS Microbiol Ecol 52:153–162
Dobbelaere S, Croonenborgh A, Thys A, Ptacek D, Okon Y, Vanderleyden J (2002) Effect of
inoculation with wild type Azospirillum brasilense and A. irakense strains on development and
nitrogen uptake of spring wheat and grain maize. Biol Fertil Soils 36:284–297
Dominguez-Nunez JA, Benito B, Berrocal-Lobo M, Albanesi A (2016) Mycorrhizal fungi: role
in the solubilization of potassium. In: Meena VS, Maurya BR, Verma JP, Meena RS (eds)
2 Rhizosphere Microorganisms Towards Soil Sustainability and Nutrient Acquisition 43
Kao PH, Huang CC, Hseu ZY (2006) Response of microbial activities to heavy metals in a neutral
loamy soil treated with biosolid. Chemosphere 64:63–70
Kavino M, Harish S, Kumar N, Saravanakumar D, Samiyappan R (2008) Induction of systemic
resistance in banana (Musa spp.) against banana bunchy top virus (BBTV) by combining chitin
with root-colonizing Pseudomonas fluorescens strain CHA0. Eur J Plant Pathol 120:353–362
Kavino M, Harish S, Kumar N, Saravanakumar D, Samiyappan R (2010) Effect of chitinolytic
PGPR on growth, yield and physiological attributes of banana (Musa spp.) under field condi-
tions. Appl Soil Ecol 45:71–77
Kingsley MT, Fredrickson JK, Metting FB et al (1994) Environmental restoration using
plant-microbe bioaugmentation. In: Hinchee RE, Leeson A, Semprini L, Ong SK (eds)
Bioremediation of chlorinated and polyaromatic hydrocarbon compounds. Lewis Publishers,
Boca Raton, pp 287–292
Kuiper I, Bloemberg GV, Lugtenberg BJJ (2001) Selection of a plant-bacterium pair as a novel
tool for rhizostimulation of polycyclic aromatic hydrocarbon-degrading bacteria. Mol Plant
Microbe Interact 14:1197–1205
Kumar A, Bahadur I, Maurya BR, Raghuwanshi R, Meena VS, Singh DK, Dixit J (2015) Does
a plant growth-promoting rhizobacteria enhance agricultural sustainability? J Pure Appl
Microbiol 9:715–724
Kumar A, Meena R, Meena VS, Bisht JK, Pattanayak A (2016a) Towards the stress management
and environmental sustainability. J Clean Prod 137:821–822
Kumar A, Patel JS, Bahadur I, Meena VS (2016b) The molecular mechanisms of KSMs for
enhancement of crop production under organic farming. In: Meena VS, Maurya BR, Verma JP,
Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture. Springer,
India, pp 61–75. doi:10.1007/978-81-322-2776-2_5
Kumar A, Maurya BR, Raghuwanshi R, Meena VS, Islam MT (2017a) Co-inoculation with
Enterobacter and Rhizobacteria on yield and nutrient uptake by wheat (Triticum aestivum L.)
in the alluvial soil under indo-gangetic plain of India. J Plant Growth Regul. doi:10.1007/
s00344-016-9663-5
Kumar A, Meena VS, Maurya BR, Raghuwanshi R, Bisht JK, Pattanayak A (2017b) Towards the
biological nitrogen fixation and nitrogen management in legume under sustainable agriculture.
Appl Soil Ecol. doi:10.1016/j.apsoil.2017.05.013
Kuzyakov Y, Xu X (2013) Competition between roots and microorganisms for nitrogen: mecha-
nisms and ecological relevance. New Phytol 198:656–669
Liste HHM (2000) Plant-promoted pyrene degradation in soil. Chemosphere 40:7–10
Lugtenberg B (2015) Life of microbes in the rhizosphere. In: Lugtenberg B (ed) Principles of plant-
microbe interactions. Springer International Publishing Switzerland, Heidelberg, pp 7–15
Marschner P, Rengel Z (2007) Nutrient cycling in terrestrial ecosystems. Springer, Berlin.
(AgEng96, Madrid, 23–26 September 1996)
Marschner P, Fu QL, Rengel Z (2003) Manganese availability and microbial populations in the
rhizosphere of wheat genotypes differing in tolerance to Mn deficiency. J Plant Nutr Soil Sci
166:712–718
Martinez-Viveros O, Jorquera MA, Crowley DE, Gajardo G, Mora ML (2010) Mechanisms and
practical considerations involved in plant growth promotion by rhizobacteria. J Soil Sci Plant
Nutr 10:293–319
Masood S, Bano A (2016) Mechanism of potassium solubilization in the agricultural soils by
the help of soil microorganisms. In: Meena VS, Maurya BR, Verma JP, Meena RS (eds)
Potassium solubilizing microorganisms for sustainable agriculture. Springer, India, pp 137–
147. doi:10.1007/978-81-322-2776-2_10
Maurya BR, Meena VS, Meena OP (2014) Influence of inceptisol and alfisol’s potassium solubiliz-
ing bacteria (KSB) isolates on release of K from waste mica. Vegetos 27:181–187
Meena OP, Maurya BR, Meena VS (2013a) Influence of K-solubilizing bacteria on release of
potassium from waste mica. Agric Sustain Dev 1:53–56
2 Rhizosphere Microorganisms Towards Soil Sustainability and Nutrient Acquisition 45
Meena VS, Maurya BR, Bohra JS, Verma R, Meena MD (2013b) Effect of concentrate manure
and nutrient levels on enzymatic activities and microbial population under submerged rice in
alluvium soil of Varanasi. Crop Res 45(1,2 & 3):6–12
Meena VS, Maurya BR, Verma R, Meena RS, Jatav GK, Meena SK, Meena SK (2013c) Soil
microbial population and selected enzyme activities as influenced by concentrate manure and
inorganic fertilizer in alluvium soil of Varanasi. The Bioscan 8(3):931–935
Meena VS, Maurya BR, Bahadur I (2014a) Potassium solubilization by bacterial strain in waste
mica. Bangladesh J Bot 43:235–237
Meena VS, Maurya BR, Verma JP (2014b) Does a rhizospheric microorganism enhance K+ avail-
ability in agricultural soils? Microbiol Res 169:337–347
Meena RS, Meena VS, Meena SK, Verma JP (2015a) The needs of healthy soils for a healthy
world. J Clean Prod 102:560–561
Meena RS, Meena VS, Meena SK, Verma JP (2015b) Towards the plant stress mitigate the agricul-
tural productivity: a book review. J Clean Prod 102:552–553
Meena VS, Maurya BR, Meena RS (2015c) Residual impact of well grow formulation and NPK on
growth and yield of wheat (Triticum aestivum L.) Bangladesh J Bot 44(1):143–146
Meena VS, Maurya BR, Verma JP, Aeron A, Kumar A, Kim K, Bajpai VK (2015d) Potassium
solubilizing rhizobacteria (KSR): isolation, identification, and K-release dynamics from waste
mica. Ecol Eng 81:340–347
Meena VS, Meena SK, Verma JP, Meena RS, Ghosh BN (2015e) The needs of nutrient use efficiency
for sustainable agriculture. J Clean Prod 102:562–563. doi:10.1016/j.jclepro.2015.04.044
Meena VS, Verma JP, Meena SK (2015f) Towards the current scenario of nutrient use efficiency in
crop species. J Clean Prod 102:556–557. doi:10.1016/j.jclepro.2015.04.030
Meena RK, Singh RK, Singh NP, Meena SK, Meena VS (2016a) Isolation of low temperature sur-
viving plant growth-promoting rhizobacteria (PGPR) from pea (Pisum sativum L.) and docu-
mentation of their plant growth promoting traits. Biocatal Agric Biotechnol 4:806–811
Meena RS, Bohra JS, Singh SP, Meena VS, Verma JP, Verma SK, Sihag SK (2016b) Towards the
prime response of manure to enhance nutrient use efficiency and soil sustainability a current
need: a book review. J Clean Prod 112(1):1258–1260
Meena SK, Rakshit A, Meena VS (2016c) Effect of seed bio-priming and N doses under varied
soil type on nitrogen use efficiency (NUE) of wheat (Triticum aestivum L.) under greenhouse
conditions. Biocatal Agric Biotechnol 6:68–75
Meena VS, Bahadur I, Maurya BR, Kumar A, Meena RK, Meena SK, Verma JP (2016d) Potassium-
solubilizing microorganism in evergreen agriculture: an overview. In: Meena VS, Maurya BR,
Verma JP, Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture.
Springer, India, pp 1–20. doi:10.1007/978-81-322-2776-2_1
Meena VS, Meena SK, Bisht JK, Pattanayak A (2016e) Conservation agricultural practices in
sustainable food production. J Clean Prod 137:690–691
Meharg AA, Cairney JWG (2000) Ectomycorrhizas – extending the capabilities of rhizosphere
remediation? Soil Biol Biochem 32:1475–1484
Mendes R, Kruijt M, de Bruijn I, Dekkers E, van der Voort M, Schneider JH, Piceno YM, DeSantis
TZ, Andersen GL, Bakker PA, Raaijmakers JM (2011) Deciphering the rhizosphere microbi-
ome for disease-suppressive bacteria. Science 332(6033):1097–1100
Meyer J, Schmidt A, Michalke K, Hensel R (2007) Volatilisation of metals and metalloids by the
microbial population of an alluvial soil. Syst Appl Microbiol 30:229–238
Mirsal I (2004) Soil pollution: origin, monitoring and remediation. Springer, New York
Morgan JAW, Bending GD, White PJ (2005) Biological costs and benefits to plant–microbe inter-
actions in the rhizosphere. J Exp Bot 56:1729–1739
Morrissey JP, Dow JM, Mark GL, O’Gara F (2004) Are microbes at the root of a solution to world
food production? EMBO Rep 5(10):922–926
Murali O, Mehar SK (2014) Bioremediation of heavy metals using spirulina. Int J Geol Earth
Environ Sci 4(1):244–249
46 M. Prasad et al.
Murphy JF, Zehnder GW, Schuster DJ, Sikora EJ, Polston JE, Kloepper JW (2000) Plant growth-
promoting rhizobacterial mediated protection in tomato against tomato mottle virus. Plant Dis
84:779–784
Nath D, Maurya BR, Meena VS (2017) Documentation of five potassium- and phosphorus-
solubilizing bacteria for their K and P-solubilization ability from various minerals. Biocatal
Agric Biotechnol 10:174–181
Naureen Zakira P, Hafeez Adam H, Roberts Fauzia Y, Michael R (2009) Identification of rice blast
disease-suppressing bacterial strains from the rhizosphere of rice grown in Pakistan. Crop Prot
28:1052–1060
Nezarat S, Gholami A (2009) Screening plant growth promoting rhizobacteria for improving seed
germination, seedling growth and yield of maize. Pak J Biol Sci 12:26–32
Nihorimbere V, Ongena M, Smargiassi M, Thonart P (2011) Beneficial effect of the rhizosphere
microbial community for plant growth and health. Biotechnol Agron Soc 15:327–337
Niranjan SR, Deepak SA, Basavaraju P, Shetty HS, Reddy MS, Kloepper JW (2003) Comparative
performance of formulations of plant promoting rhizobacteria in growth promotion and sup-
pression of downy mildew in pearl millet. Crop Prot 22:579–588
Noel TC, Sheng C, Yost CK, Pharis RP, Hynes MF (1996) Rhizobium leguminosarum as a plant
growth-promoting rhizobacterium: direct growth promotion of canola and lettuce. Can
J Microbiol 42:279–283
Palada MC, Wu DL (2005) Influence of rain shelter and irrigation method on yield, water and
fertilizer use efficiency of chili pepper. Hortic Sci 40:1143. (abst.)
Parewa HP, Yadav J, Rakshit A, Meena VS, Karthikeyan N (2014) Plant growth promoting rhizo-
bacteria enhance growth and nutrient uptake of crops. Agric Sustain Dev 2(2):101–116
Patil SS, Adetutu EM, Rochow J, Mitchell JG, Ball AS (2014) Sustainable remediation: electro-
chemically assisted microbial dechlorination of tetrachloroethene-contaminated groundwater.
Microb Biotechnol 7(1):54–63
Peix A, Rivas-Boyero AA, Mateos PF, Rodriguez-Barrueco C, Martínez-Molina E, Velazquez
E (2001) Growth promotion of chickpea and barley by a phosphate solubilizing strain of
Mesorhizobium mediterraneum under growth chamber conditions. Soil Biol Biochem
33:103–110
Postel S, Polak P, Gonzales F, Keller J (2003) Drip irrigation for small farmers: a new initiative to
alleviate hunger and poverty. International Development Enterprises (IDE-India) Report 014
Prakash S, Verma JP (2016) Global perspective of potash for fertilizer production. In: Meena VS,
Maurya BR, Verma JP, Meena RS (eds) Potassium solubilizing microorganisms for sustainable
agriculture. Springer, India, pp 327–331. doi:10.1007/978-81-322-2776-2_23
Priyadharsini P, Muthukumar T (2016) Interactions between arbuscular mycorrhizal fungi and
potassium-solubilizing microorganisms on agricultural productivity. In: Meena VS, Maurya
BR, Verma JP, Meena RS (eds) Potassium solubilizing microorganisms for sustainable agricul-
ture. Springer, India, pp 111–125. doi:10.1007/978-81-322-2776-2_8
Qiu X, Shah SJ, Kendall EW et al (1994) Grass enhanced bioremediation for clay soils contaminated
with polynuclear aromatic hydrocarbons. In: Anderson TA, Coates JR (eds) Bioremediation
through rhizosphere technology. American Chemical Society, Washington, DC, pp 142–157
Radjacommare R, Nandakumar R, Kandan A, Suresh S, Bharathi M, Raguchander T, Samiyappan
R (2002) Pseudomonas fluorescens based bio-formulation for the management of sheath blight
disease and leaf folder insect in rice. Crop Prot 21:671–677
Radvan S, Sorkhoh N, El Nemr L (1995) Oil biodegradation around roots. Nature:376–302
Raghavendra MP, Nayaka NC, Nuthan BR (2016) Role of rhizosphere microflora in potas-
sium solubilization. In: Meena VS, Maurya BR, Verma JP, Meena RS (eds) Potassium
solubilizing microorganisms for sustainable agriculture. Springer, India, pp 43–59.
doi:10.1007/978-81-322-2776-2_4
Rawat J, Sanwal P, Saxena J (2016) Potassium and its role in sustainable agriculture. In: Meena
VS, Maurya BR, Verma JP, Meena RS (eds) Potassium solubilizing microorganisms for sus-
tainable agriculture. Springer, India, pp 235–253. doi:10.1007/978-81-322-2776-2_17
2 Rhizosphere Microorganisms Towards Soil Sustainability and Nutrient Acquisition 47
Richardson AE, Barea JM, McNeill AM, PrigentCombaret C (2009) Acquisition of phosphorus
and nitrogen in the rhizosphere and plant growth promotion by microorganisms. Plant Soil
321:305–339
Rodriguez Hernandez A, Heydrich Perez M, Acebo Guerrero Y, Velazquez del Valle MG,
Hernandez Lauzardo AN (2008) Antagonistic activity of Cuban native rhizobacteria against
Fusarium verticillioides (Sacc.) Nirenb. In maize (Zea mays L.). Appl Soil Ecol 39:180–186
Rokhzadi A, Ahmad A, Farrokh D, Ghorban NM, Eslam M (2008) Influence of plant growth-
promoting rhizobacteria on dry matter accumulation and yield of chickpea (Cicer arietinum
L.) under field conditions. Am Eur J Agric Environ Sci 3:253–257
Saha M, Maurya BR, Bahadur I, Kumar A, Meena VS (2016a) Can potassium-solubilising bac-
teria mitigate the potassium problems in India? In: Meena VS, Maurya BR, Verma JP, Meena
RS (eds) Potassium solubilizing microorganisms for sustainable agriculture. Springer, India,
pp 127–136. doi:10.1007/978-81-322-2776-2_9
Saha M, Maurya BR, Meena VS, Bahadur I, Kumar A (2016b) Identification and characterization
of potassium solubilizing bacteria (KSB) from Indo-Gangetic Plains of India. Biocatal Agric
Biotechnol 7:202–209
Saleem M, Arshad M, Hussain S, Bhatti AS (2007) Perspective of plant growth promoting rhizo-
bacteria (PGPR) containing ACC deaminase in stress agriculture. J Ind Microbiol Biotechnol
34:635–648
Sallih Z, Bottner P (1988) Effects of wheat (Triticum aestivum) roots on mineralization rates of soil
organic matter. Biol Fertil Soils 7:67–70
Saravanakumar D, Samiyappan R (2007) ACC deaminase from Pseudomonas fluorescens mediated
saline resistance in groundnut (Arachis hypogea) plants. J Appl Microbiol 102(5):1283–1292
Saravanakumar D, Lavanya N, Muthumeena K, Raguchander T, Samiyappan R (2009) Fluorescent
pseudomonad mixtures mediate disease resistance in rice plants against sheath rot (Sarocladium
oryzae) disease. BioControl 54:273–286
Sarkar D, Meena VS, Haldar A, Rakshit R (2017) Site-specific nutrient management (SSNM):
a unique approach towards maintaining soil health. In The adaptive soil management: from
theory to practices. p 69–88. doi:10.1007/978-981-10-3638-5_3
Schnoor JL, Aitchison EW, Kelley SL et al (1998) Phytoremediation and augmentation of
1,4-dioxane. Abstr Adv Biol Rern Syst Conf 87:91–133
Schwab AP, Banks MK, Arunachalam M (1995) Biodegradation of polyromantic hydrocarbon in
rhizosphere. In: Hinchee RE, Anderson DB, Hoeppel RE (eds) Bioremediation of recalcitrant
organics. Battle Memorial Institute, Columbus, pp 23–29
Sharma A, Shankhdhar D, Shankhdhar SC (2016) Potassium-solubilizing microorganisms:
mechanism and their role in potassium solubilization and uptake. In: Meena VS, Maurya BR,
Verma JP, Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture.
Springer, India, pp 203–219. doi:10.1007/978-81-322-2776-2_15
Shrivastava M, Srivastava PC, D’Souza SF (2016) KSM soil diversity and mineral solubilization,
in relation to crop production and molecular mechanism. In: Meena VS, Maurya BR, Verma JP,
Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture. Springer,
India, pp 221–234. doi:10.1007/978-81-322-2776-2_16
Silva HSA, da Silva RR, Macagnan M, de Almeida HVB, Pereira MCB, Mounteer A (2004)
Rhizobacterial induction of systemic resistance in tomato plants: non-specific protection and
increase in enzyme activity. Biol Control 29:288–295
Sindhu SS, Parmar P, Phour M, Sehrawat A (2016) Potassium-solubilizing microorganisms
(KSMs) and its effect on plant growth improvement. In: Meena VS, Maurya BR, Verma JP,
Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture. Springer,
India, pp 171–185. doi:10.1007/978-81-322-2776-2_13
Singh YV, Mandal BK (2000) Rate of mineralization of Azolla, other organic materials and urea in
water logged soils. Trop Agric (Trinidad) 77(1):119–122
Singh BK, Millard P, Whiteley AS, Murrell JC (2004) Unravelling rhizosphere–microbial interac-
tions: opportunities and limitations. Trends Microbiol 12(8):386–393
48 M. Prasad et al.
Singh NP, Singh RK, Meena VS, Meena RK (2015) Can we use maize (Zea mays) rhizobacteria as
plant growth promoter? Vegetos 28(1):86–99. doi:10.5958/2229-4473.2015.00012.9
Singh M, Dotaniya ML, Mishra A, Dotaniya CK, Regar KL, Lata M (2016) Role of biofertil-
izers in conservation agriculture. In: Bisht JK, Meena VS, Mishra PK, Pattanayak A (eds)
Conservation agriculture: an approach to combat climate change in Indian Himalaya. Springer,
Singapore, pp 113–134. doi:10.1007/978-981-10-2558-7_4
Smith SE, Facelli F, Pope S, Smith FA (2010) Plant performance in stressful environments: interpret-
ing new and established knowledge of the roles of arbuscular mycorrhizas. Plant Soil 326:3–20
Tebo BM, Bargar JR, Clement BG, Dick GJ, Murray KJ, Parker D, Verity R, Webb SM (2004)
Biogenic manganese oxides: properties and mechanisms of formation. Ann Rev Earth Planet
Sci 32:287–328
Teotia P, Kumar V, Kumar M, Shrivastava N, Varma A (2016) Rhizosphere microbes: potassium
solubilization and crop productivity-present and future aspects. In: Meena VS, Maurya BR,
Verma JP, Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture.
Springer, India, pp 315–325. doi:10.1007/978-81-322-2776-2_22
Umrania VV (2006) Bioremediation of toxic heavy metals using acid thermophilic autotrophs.
Bioresour Technol 97:1237–1242
Vaxevanidou K, Christou C, Kremmydas GF, Georgakopoulos DG, Papassiopi N (2015) Role of
indigenous arsenate and iron (III) respiring microorganisms in controlling the mobilization of
arsenic in a contaminated soil sample. Bull Environ Contam Toxicol 94(3):282–288
Velazquez E, Silva LR, Ramírez-Bahena MH, Peix A (2016) Diversity of potassium-solubilizing
microorganisms and their interactions with plants. In: Meena VS, Maurya BR, Verma JP,
Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture. Springer,
India, pp 99–110. doi:10.1007/978-81-322-2776-2_7
Verma R, Maurya BR, Meena VS (2014) Integrated effect of bio-organics with chemical fertilizer
on growth, yield and quality of cabbage (Brassica oleracea var capitata). Indian J Agric Sci
84(8):914–919
Verma JP, Jaiswa DK, Meena VS, Meena RS (2015a) Current need of organic farming for enhanc-
ing sustainable agriculture. J Clean Prod 102:545–547
Verma JP, Jaiswal DK, Meena VS, Kumar A, Meena RS (2015b) Issues and challenges about sus-
tainable agriculture production for management of natural resources to sustain soil fertility and
health. J Clean Prod 107:793–794
Verma R, Maurya BR, Meena VS, Dotaniya ML, Deewan P (2017a) Microbial dynamics as
influenced by bio-organics and mineral fertilizer in alluvium soil of Varanasi. India Int J Curr
Microbiol Appl Sci 6(2):1516–1524
Verma R, Maurya BR, Meena VS, Dotaniya ML, Deewan P, Jajoria M (2017b) Enhancing pro-
duction potential of cabbage and improves soil fertility status of Indo-Gangetic plain through
application of bio-organics and mineral fertilizer. Int J Curr Microbiol App Sci 6(3):301–309
Weber KA, Achenbach LA, Coates JD (2006) Microorganisms pumping iron: anaerobic microbial
iron oxidation and reduction. Nat Rev Microbiol 4:752–764
Welbaum G, Sturz AV, Dong Z, Nowak J (2004) Fertilizing soil microorganisms to improve pro-
ductivity of agroecosystems. Crit Rev Plant Sci 23:175–193
Whitman WB, Coleman DC, Wiebe WJ (1998) Prokaryotes: the unseen majority. Proc Natl Acad
Sci U S A 95:6578–6583
Wood N (2001) Nodulation by numbers: the role of ethylene in symbiotic nitrogen fixation. Trends
Plant Sci 6:501–502
Wu SC, Cao ZH, Li ZG, Cheung KC, Wong MH (2005) Effects of biofertilizer containing N-fixer, P
and K solubilizers and AM fungi on maize growth: a greenhouse trial. Geoderma 125:155–166
Yadav BK, Sidhu AS (2016) Dynamics of potassium and their bioavailability for plant nutrition.
In: Meena VS, Maurya BR, Verma JP, Meena RS (eds) Potassium solubilizing microorganisms
for sustainable agriculture. Springer, India, pp 187–201. doi:10.1007/978-81-322-2776-2_14
Yang C, Yang L, Yang Y, Ouyyang Z (2004) Rice root growth and nutrient uptake as influenced
by organic manure in continuously and alternately flooded paddy soils. Agric Water Manag
70:67–81
2 Rhizosphere Microorganisms Towards Soil Sustainability and Nutrient Acquisition 49
Yasari E, Patwardhan AM (2007) Effects of Azotobacter and Azospirillum inoculations and chemi-
cal fertilizers on growth and productivity of Canola. Asian J Plant Sci 6:77–82
Yasin M, Munir I, Faisal M (2016) Can Bacillus spp. enhance K+ uptake in crop species. In: Meena
VS, Maurya BR, Verma JP, Meena RS (eds) Potassium solubilizing microorganisms for sus-
tainable agriculture. Springer, India, pp 163–170. doi:10.1007/978-81-322-2776-2_12
Zahedi H (2016) Growth-promoting effect of potassium-solubilizing microorganisms on
some crop species. In: Meena VS, Maurya BR, Verma JP, Meena RS (eds) Potassium
solubilizing microorganisms for sustainable agriculture. Springer, India, pp 31–42.
doi:10.1007/978-81-322-2776-2_3
Zang S, Moyne AL, Redddy MS, Kloepper JW (2002) The role of salicylic acid in induced sys-
temic resistance elicited by plant growth-promoting rhizobacteria against blue mold of tobacco.
Biol Control 25:288–296
Zehnder GW, Yao C, Murphy JF, Sikora EJ, Kloepper JW (2000) Induction of resistance in tomato
against cucumber mosaic cucumovirus by plant growth promoting rhizobacteria. Biol Control
45:127–137
Zehnder G, Murphy JF, Sikora EJ, Kloepper JW (2001) Application of rhizobacteria for induced
resistance. Eur J Plant Pathol 107:39–50
Zhang S, White TL, Martinez M, McInroy C, Kloepper JA, Klassen JWW (2010) Evaluation of
plant growth-promoting rhizobacteria for control of Phytophthora blight on squash under
greenhouse conditions. Biol Control 53:129–135
Zhang FS, Cui ZL, Chen XP, Ju XT, Shen JB, Chen Q, Liu XJ, Zhang WF, Mi GH, Fan MS
(2012) Integrated nutrient management for food security and environmental quality in China.
In: Sparks DL (ed) Advances in agronomy, vol 116. Academic, San Diego, pp 1–40
Zhang A, Zhao G, Gao T, Wang W, Li J, Zhang S et al (2013) Solubilization of insoluble potassium
and phosphate by Paenibacillus kribensis CX-7:a soil microorganism with biological control
potential. Afr J Microbiol Res 7(1):41–47
Zhao Q, Zeng D, Fan Z (2010) Nitrogen and phosphorus transformations in the rhizospheres of
three tree species in a nutrient-poor sandy soil. Appl Soil Ecol 46:341–346
Zolla G, Badri DV, Bakker MG, Manter DK, Vivanco JM (2013) Soil microbiomes vary in their
ability to confer drought tolerance to Arabidopsis. Appl Soil Ecol 68:1–9
PGPR: Heart of Soil and Their Role in Soil
Fertility 3
Ishwar Prakash Sharma, Satish Chandra, Narendra Kumar,
and Dinesh Chandra
Abstract
Soil is an excellent niche for the growth of microorganisms which includes plant
growth-promoting rhizobacteria (PGPRs). PGPRs have great potential for plant
growth promotion as they control pest and disease and have been considered
important in sustainable agriculture. PGPRs induce and/or synthesize various
growth-promoting and biocontrolling chemicals which are useful for sustainable
agriculture and environment. These chemicals are phytohormones, antibiotics,
enzymes, etc., which react with the soil and provide better health to the plants.
PGPRs directly or indirectly circulate their products to soil for better soil health
which further enhance the soil immunity followed by influencing plant growth,
productivity, and yield. Soil immune system works strongly by the activities of
soil microbes which are considered as a heart of soil. These soil microbes are
responsible for increasing soil fertility and productivity which are considered
important for sustainable agriculture.
Keywords
PGPR • Immunity • Sustainable agriculture • Soil fertility • Biomolecules
3.1 Introduction
Soil is an excellent niche for the growth of microorganisms including bacteria. Out
of these some bacteria are able to colonize soil surrounding plant roots, rhizosphere
(Hiltner 1904; Kennedy 2005); these bacteria are rhizobacteria. These bacteria have
the ability to induce plant growth and biocontrol activities (Meena et al. 2016e). In
the coming years, food security and feed will be a major global challenge for
increasing human population; now, we need to increase the sustainability of agricul-
tural productivity to meet the increasing demand for food. More than half of the
population still relies on agriculture as its principal source of income, and it is a
source of raw material for a large number of industries (Meena et al. 2013a; Bahadur
et al. 2014; Maurya et al. 2014; Jat et al. 2015; Zahedi 2016; Kumar et al. 2017).
Therefore, these bacteria are an important biological tool for the abovementioned
challenges.
Agricultural crops are mostly affected by diseases and/or pests which can be
biologically prevented by these PGPRs and other symbiotic rhizospheric microor-
ganisms which are mostly found in the rhizosphere of all plants (Trudgill and Blok
2001; Sharma and Sharma 2016). For the managing of pest and disease, a large
number of synthetic chemicals are being used, which pose environmental hazards,
besides being costly and uneconomical (Joshi et al. 2006; Maheshwari et al. 2012;
Youssef and Eissa 2014; Gupta et al. 2015). The quantity and quality of agricultural
production are affected by the use of chemicals in agriculture practices; recently,
these interactions are an important discussion subject under the agriculture-
environment relationships (Ladha et al. 1997; Eser and Geçit 2010; Gupta et al.
2015; Meena et al. 2016f; Mahmood et al. 2016a, b; Ahmad et al. 2016a, b; Sharma
and Sharma 2016). PGPRs are used to replace these chemicals by a variety of mech-
anisms, viz., formation of soil structure, decomposition of organic matter, recycling
of elements, mineral nutrient solubilization, production of plant growth regulators,
degradation of organic pollutants, root growth stimulation, soil fertility enhance-
ment, biocontrol of plant pathogens, etc. (Ahemad and Khan 2009; Akhtar et al.
2012; Sivasakhti et al. 2014; Gupta et al. 2015).
Many studies show that the use of microbial-based products is a common prac-
tice for controlling plant pest population and preventing diseases of crops (Lacey
et al. 2001; Goettel et al. 2010; Dogan et al. 2016; Meena et al. 2013c, 2015e). The
major influences that the rhizosphere microorganisms have on plants today become
an important biological tool to govern the health of plants in eco-friendly manner.
They are involved in various biotic activities of the soil ecosystem to make it
dynamic for nutrient turnover and sustainable for crop production (Ahemad and
Khan 2009; Akhtar et al. 2012; Gupta et al. 2015). Hence, PGPRs are economic and
secure alternative strategy for enhancing plant growth and development as they are
able to increase the soil health and immunity toward soil pests and diseases.
3 PGPR: Heart of Soil and Their Role in Soil Fertility 53
Soil is the natural niche for soil microorganisms which play an important role in soil
processes and determine plant productivity (Meena et al. 2014b, 2015c, d). PGPRs
are involved in all beneficial activities within the soil like decomposition of crop
residues, synthesis of soil organic matter (SOM), mineralization of SOM, immobi-
lization of mineral nutrients, phosphate solubilization, nitrification, nitrogen fixa-
tion, phytohormone synthesis, and disease suppression which help in soil fertility
improvement and ultimately enhancing the productivity (Prasad et al. 2015). Soil
moisture is a necessary condition for PGPR colonization in the rhizosphere
(Shrivastava et al. 2014). Rhizobacteria associated with roots are responsible for the
synthesis of large number of biomolecules which get mixed with the soil and further
improve soil health. They also act on the plant residues, where various organic com-
pounds undergo decomposition and mineralization (Kumar et al. 2015; Ahmad
et al. 2016a, b; Bahadur et al. 2016a; Verma et al. 2014, 2015b; Yadav and Sidhu
2016; Yasin et al. 2016). These mineralized chemicals enter the soil body where
they agitate and add value to the soil. Apart from this mechanism, they also synthe-
size many growth-responsible chemicals which ultimately affect plant root mor-
phology (Fig. 3.1).
plant root cells, PGPRs can be classified into extracellular plant growth-promoting
rhizobacteria (ePGPR) and intracellular plant growth-promoting rhizobacteria
(iPGPR) (Gray and Smith 2005; Meena et al. 2015a, 2016b; Raghavendra et al.
2016; Rawat et al. 2016; Saha et al. 2016a). iPGPRs can enter plant cells and are
able to produce specialized structures, so-called nodules. On the other hand, ePG-
PRs are found in the rhizosphere, or within the apoplast of the root cortex, but not
inside the cells. According to their close proximity to the plant root, ePGPRs can be
divided into three classes, i.e., living near the roots but not in contact, colonizing on
the root surface, and living in the intercellular spaces of the root cortex (Gray and
Smith 2005; Dotaniya et al. 2016; Jaiswal et al. 2016; Jha and Subramanian 2016;
Kumar et al. 2016a, b; Masood and Bano 2016; Singh et al. 2015, 2016).
PGPRs act against phytopathogens by secretion of many biochemicals like toxin,
enzymes, proteins, antibiotics, etc. Plant-associated microorganisms have been
extensively examined for their role in natural and induced suppressiveness of soil-
borne diseases. Applications of PGPRs have been reported to increase the produc-
tion of antibiotics and inhibit root penetration of phytopathogens (Handelsman et al.
1990; Sikora 1992; Sikora and Hergarten 1992; Liu and Sinclair 1992; Yu et al.
2002).
PGPRs are classified on the basis of their locations, functions, and activities.
PGPRs
Intracellular Phytostimulators
PGPRs Biocontroling bacteria
Rhizoremediators
Stress homeoregulating
bacteria
Biopesticides
Bioprotectants
56 I.P. Sharma et al.
Biocontrolling Bacteria
Bashan et al. (1993) for the first time suggested these bacteria. They indirectly pro-
mote plant growth by releasing phytotoxic substances such as hydrogen cyanide,
chitinase, pectinase, cellulase, ethylene, antibiotics, etc., which are responsible for
controlling plant pathogens (Ramamoorthy and Samiyappan 2001; Yu et al. 2002;
Bashan and de-Bashan 2010; Bhattacharyya and Jha 2012; Gupta et al. 2015;
Sharma and Sharma 2016; Meena et al. 2016) (Table 3.2).
Table 3.2 Some PGPRs and their antibiotic production and target pathogen (Bhattacharyya and
Jha 2012)
Antibiotics PGPRs Target organisms
Bacillomycin Bacillus Aspergillus flavus
Kanosamine B. cereus Phytophthora medicaginis
Zwittermycin A B. cereus, B. Phytophthora, Sclerotinia sclerotiorum
thuringiensis, B. subtilis
Iturin B. subtilis Pythium ultimum, Rhizoctonia solani,
Fusarium oxysporum
Iturin A, surfactin B. subtilis Rhizoctonia solani
Pyrrolnitrin Burkholderia cepacia, Rhizoctonia solani, Gaeumannomyces
Pseudomonas fluorescens, graminis var. tritici, Agrobacterium
Enterobacter tumefaciens, Clavibacter michiganensis,
agglomerans Xanthomonas campestris, Pseudomonas
syringae
2,4-DAPG, Pseudomonas fluorescens Xanthomonas oryzae, Pythium ultimum,
pyoluteorin, Gaeumannomyces graminis var. tritici
phenazines
Phenazine-1- Pseudomonas fluorescens, Gaeumannomyces graminis var. tritici,
carboxylate P. aureofaciens Sclerotinia homeocarpa
Phenazine-1- P. chlororaphis Fusarium oxysporum f. sp.
carboxamide radicis-lycopersici
Viscosinamide Pseudomonas fluorescens R. solani, Pythium ultimum
Amphisin Pseudomonas fluorescens Pythium ultimum, R. solani
Geldanamycin A Streptomyces R. solani
hygroscopicus
Oligomycin A Streptomyces libani Botrytis cinerea
Polyoxin D Streptomyces cacaoi R. solani
58 I.P. Sharma et al.
(A) Biofertilizers – Many of the PGPRs enhance soil fertility and productivity by
involving in nutrient cycling. Different types of biofertilizers are being used
such as:
1.
Nitrogen-fixing biofertilizers like Rhizobium, Bradyrhizobium,
Nitrosomonas, Azospirillum, Azotobacter, etc.
2. Phosphorus-solubilizing and phosphorus-mobilizing biofertilizers like
Bacillus, Pseudomonas, Mycorrhiza, etc.
3. Plant growth-promoting biofertilizers like Pseudomonas, Bacillus, etc.
(B) Phytostimulators – Certain bacteria stimulate the plant growth by producing dif-
ferent phytohormones like Pseudomonas, Azospirillum, Bradyrhizobium, etc.
(C) Rhizoremediators – Many of the bacteria are used in degradation of herbicides,
pesticides, and other hazardous organic compounds and/or pollutants in the
soil which could be helpful in plant growth by increasing soil fertility.
(D) Biopesticides – Many of the bacteria produce antibiotics and other toxic com-
pounds which are helpful in the reduction of pathogens, e.g., Pseudomonas and
Bacillus spp.
(E) Bioprotectants – Large numbers of bacteria are helpful in the disease suppres-
sion which directly and indirectly enhance plant growth.
PGPRs promote plant growth directly as well as indirectly (Castro et al. 2009) as
follows:
Phosphate
Cyanide compounds
solubilization
Potassium Polysaccharide
solubilization production
Hydrolytic enzymes
Siderophore production
production
Directly, PGPRs promote plant growth by providing essential nutrients (N, P, Fe,
etc.) and enhancing phytohormone level. Generally these bacteria are involved in
different nutrient cycles like N2 and P cycle; directly the nutrients are provided to
plants by these bacteria through nutrient mineralization and solubilization pro-
cesses. On the other hand, these bacteria are also involved in many phytohormone
(IAA, gibberellic acid, cytokinins) pathways (Kloepper et al. 2007). Among all the
phytohormones, IAA is the most common which is produced by PGPR (Barazani
and Friedman 1999).
PGPRs are the well source for biocontrol which inhibit the phytopathogen, so they
are indirectly helpful to plant growth (Glick 2012). PGPRs are powerful resource of
disease suppression which are economically as well as environmentally safe. PGPRs
have a variety of mechanisms which make them a biocontrol agent. They produce
antibiotics (Mazurier et al. 2009), cell wall-degrading enzymes like cellulose and
β-1,3-glucanase, and cyanide compounds like HCN (Duffy et al. 2003; Ramatte
et al. 2003) which are helpful in reducing phytopathogens and enhancing plant
growth. Some molecules such as 2, 4-diacetylphloroglucinol (2, 4-DAPG) and pyo-
luteorin are also produced by some PGPRs (de Souza et al. 2003; Beneduzi et al.
2012) which are responsible for disease suppression.
By using the above mechanisms, PGPRs defense against phytopathogens and
pests results in resistance to the plant. This resistance is either systemic acquired
resistance (SAR) or induced systemic resistance (ISR). SAR is activated in plant
tissues after pathogen attacks, immediately plant signals are produced, and defense
responses occur in distal tissues through the vascular system (Pieterse et al. 2009).
As the resistance induced, the plant is protected from pathogenic fungi, bacteria,
viruses and nematodes. A large number of defense enzymes like phenylalanine
ammonia lyase (PAL), chitinase, β-1,3-glucanase, peroxidase (PO), polyphenol oxi-
dase (PPO), lipoxygenase (LOX), superoxide dismutase (SOD), catalase (CAT),
and ascorbate peroxidase (APX) are involved in the resistance and activate ISR
(Meena et al. 2016a; Saha et al. 2016b; Sharma et al. 2016; Shrivastava et al. 2016;
Sindhu et al. 2016).
3.5 Conclusions
Several microbes from soil have been studied and are responsible for the improve-
ment of soil fertility and productivity, crop production and protection, and compost
preparation and decomposition. The products from these soil bacteria have been
exploited to control many diseases. Recent development of molecular biology made
it possible to develop transgenic plants by improving genetics. Similarly, the
60 I.P. Sharma et al.
Acknowledgments The authors are greatly thankful to all the supporting members of the
GBPUAT Pantnagar who encourage us to prepare this manuscript.
References
Ahemad M, Khan MS (2009) Effect of insecticide-tolerant and plant growth promoting
Mesorhizobium on the performance of chickpea grown in insecticide stressed alluvial soils.
J Crop Sci Biotechnol 12:213–222
Ahemad M, Khan MS (2012) Effects of pesticides on plant growth promoting traits of
Mesorhizobium strain MRC4. J Saudi Soc Agric Sci 11:63–71
Ahemad M, Kibret M (2014) Mechanisms and applications of plant growth promoting rhizobacte-
ria: current perspective. J King Saud Univ Sci 26(1):1–20
Ahmad M, Nadeem SM, Naveed M, Zahir ZA (2016a) Potassium-solubilizing bacteria and their
application in agriculture. In: Meena VS, Maurya BR, Verma JP, Meena RS (eds) Potassium
solubilizing microorganisms for sustainable agriculture. Springer, India, pp 293–313.
doi:10.1007/978-81-322-2776-2_21
Ahmad I, Khan MSA, Aqil F, Singh M (2016b) Microbial applications in agriculture and the
environment: a broad perspective (ed) Microbes and microbial technology: agricultural and
environmental applications, DOI 10.1007/978-1-4419-7931-5_1
Akhtar N, Qureshi MA, Iqbal A, Ahmad MJ, Khan KH (2012) Influence of Azotobacter and IAA
on symbiotic performance of Rhizobium and yield parameters of lentil. J Agric Res 50:361–372
Bahadur I, Meena VS, Kumar S (2014) Importance and application of potassic biofertilizer in
Indian agriculture. Int Res J Biol Sci 3:80–85
Bahadur I, Maurya BR, Kumar A, Meena VS, Raghuwanshi R (2016a) Towards the soil sustain-
ability and potassium-solubilizing microorganisms. In: Meena VS, Maurya BR, Verma JP,
Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture. Springer,
India, pp 225–266. doi:10.1007/978-81-322-2776-2_18
Bahadur I, Maurya BR, Meena VS, Saha M, Kumar A, Aeron A (2016b) Mineral release dynamics
of tricalcium phosphate and waste muscovite by mineral-solubilizing rhizobacteria isolated
from indo-gangetic plain of India. Geomicrobiol J. doi:10.1080/01490451.2016.1219431
Barazani O, Friedman J (1999) Is IAA the major root growth factor secreted from plant-growth-
mediating bacteria? J Chem Ecol 25(10):2397–2406
3 PGPR: Heart of Soil and Their Role in Soil Fertility 61
Bashan Y, de-Bashan LE (2010) How the plant growth-promoting bacterium Azospirillum pro-
motes plant growth – a critical assessment. Adv Agron 108:77–136
Bashan Y, Holguin G, Lifshitz R (1993) Isolation and characterization of plant growth-promoting
rhizobacteria. In: Glick BR, Thompson JE (eds) Methods in plant molecular biology and bio-
technology. CRC, Boca Raton, pp 331–345
Beneduzi A, Ambrosini A, Passaglia LM (2012) Plant growth-promoting rhizobacteria (PGPR):
their potential as antagonists and biocontrol agents. Genet Mol Biol 35:1044–1051
Bhattacharyya PN, Jha DK (2012) Plant growth-promoting rhizobacteria (PGPR): emergence in
agriculture. World J Microbiol Biotechnol 28:1327–1350
Bouizgarne B (2013) Bacteria for plant growth promotion and disease management. In: Maheshwari
DK (ed) Bacteria in agrobiology: disease management. Springer-Verlag, Berlin
Burr TJ, Schroth MN, Suslow T (1978) Increased potato yields by treatment of seedpieces with
specific strains of Pseudomonas fluorescens and P. putida. Phytopathology 68:1377–1383
Cassán F, Perrig D, Sgroy V, Masciarelli O, Penna C, Luna V (2009) Azospirillum brasilenseAz39
and Bradyrhizobium japonicum E109, inoculated singly or in combination, promote seed ger-
mination and early seedling growth in corn (Zea mays L.) and soybean (Glycine max L.) Eur
J Soil Biol 45:28–35
Castro RO, Cornejo HAC, Rodriguez LM, Bucio JL (2009) The role of microbial signals in plant
growth and development. Plant Signal Behav 4(8):701–712
Choudhary DK, Sharma KP, Gaur RK (2011) Biotechnological perspectives of microbes in agro-
ecosystems. Biotechnol Lett 33:1905–1910
Cohen AC, Travaglia CN, Bottini R, Piccoli PN (2009) Participation of abscisic acid and gibberel-
lins produced by endophytic Azospirillum in the alleviation of drought effects in maize. Botany
87:455–462
Cohen AC, Bottini R, Pontin M, Berli FJ, Moreno D, Boccanlandro H, Travaglia CN, Piccoli PN
(2015) Azospirillum brasilense ameliorates the response of Arabidopsis Thaliana to drought
mainly via enhancement of ABA levels. Physiol Plant 153:79–90
Das I, Pradhan M (2016) Potassium-solubilizing microorganisms and their role in enhancing
soil fertility and health. In: Meena VS, Maurya BR, Verma JP, Meena RS (eds) Potassium
solubilizing microorganisms for sustainable agriculture. Springer, India, pp 281–291.
doi:10.1007/978-81-322-2776-2_20
de Souza JTA, Arnould C, Deulvot C, Lemanceau P, Gianinazzi-Pearson V, Raaijmakers JM (2003)
Effect of 2,4-diacetylphloroglucinol on Pythium: cellular responses and variation in sensitivity
among propagules and species. Phytopathology 93:966–975
Dodd IC, Zinovkina NY, Safronova VI, Belimov AA (2010) Rhizobacterial mediation of plant
hormone status. Ann Appl Biol 157:361–379
Dogan A, Erler F, Erkan M, Ozgur Ates A, Sule Sabanci H, Polat E (2016) Microbial-based
production system: a novel approach for plant growth and Pest and disease Management in
Greenhouse-Grown Peppers (Capsicum annuum L.) J Agri Sci Technol 18:371–386
Dominguez-Nunez JA, Benito B, Berrocal-Lobo M, Albanesi A (2016) Mycorrhizal fungi: role
in the solubilization of potassium. In: Meena VS, Maurya BR, Verma JP, Meena RS (eds)
Potassium solubilizing microorganisms for sustainable agriculture. Springer, India, pp 77–98.
doi:10.1007/978-81-322-2776-2_6
Dotaniya ML, Meena VD, Basak BB, Meena RS (2016) Potassium uptake by crops as well
as microorganisms. In: Meena VS, Maurya BR, Verma JP, Meena RS (eds) Potassium
solubilizing microorganisms for sustainable agriculture. Springer, India, pp 267–280.
doi:10.1007/978-81-322-2776-2_19
Duffy B, Schouten A, Raajimakers J (2003) Pathogen selfdefense: mechanisms to counteract
microbial antagonism. Annu Rev Phytopathol 45:501–538
Eser D, Geçit H (2010) Ekoloji. A.Ü. Ziraat Fakültesi. 1584, Ders Kitab : 536
George TS, Dou D, Wang X (2016) Plant–microbe interactions: manipulating signals to enhance
agricultural sustainability and environmental security. Plant Growth Regul 80(1):1–3
Glick BR (2012) Plant growth-promoting bacteria: mechanisms and applications Hindawi publish-
ing corporation, Scientifica
62 I.P. Sharma et al.
Glick BR, Patten CL, Holguin G, Penrose GM (1999) Biochemical and genetic mechanisms used
by plant growth promoting bacteria. Imperial College Press, London
Goettel MS, Eilenberg J, Glare TR (2010) Entomopathogenic fungi and their role in regulation of
insect populations. In: Gilbert LI, Gill S (eds) Insect control: biological and synthetic agents.
Academic, London, pp 387–432
Goswami D, Janki N, Thakker DPC (2016) Portraying mechanics of plant growth promoting rhi-
zobacteria (PGPR): a review. Cogent Food Agri 2:1–19
Gray EJ, Smith DL (2005) Intracellular and extracellular PGPR: commonalities and distinctions in
the plant-bacterium signaling processes. Soil Biol Biochem 37(3):395–412
Gupta S, Meena MK, Datta S (2014) Isolation, characterization of plant growth promoting bac-
teria from the plant Chlorophytum borivilianum and in-vitro screening for activity of nitro-
gen fixation, phosphate solubilization and IAA production. Int J Cur Microbiol Appl Sci
3(7):1082–1090
Gupta G, Parihar SS, Ahirwar NK, Snehi SK, Singh V (2015) Plant growth promoting Rhizobacteria
(PGPR): current and future prospects for development of sustainable agriculture. J Microb
Biochem Technol 7(2):96–102
Handelsman J, Raffel S, Mester E, Wunderlich L, Grau C (1990) Biological control of damping off
of alfalfa seedlings with Bacillus cereus UW85. Appl Environ Microbiol 56:713–718
Hellriegel H, Wilfarth H (1888) Untersuchungen uber die Stickstoffnahrung der Gramineen und
Leguminosen. Beilageheft zu der Z des Vereins Rubenzucker-Industrie Deutschen Reichs 234
Hiltner L (1904) Uber neue erfahrungen und probleme auf dem gebiete der bodenbakteriologie.
Arb Dtsch Landwirtsch Ges 98:59–78
Hussain A, Hasnain S (2009) Cytokinin production by some bacteria: its impact on cell division in
cucumber cotyledons. Afr J Microbiol Res 3:704–712
Jaiswal DK, Verma JP, Prakash S, Meena VS, Meena RS (2016) Potassium as an important plant
nutrient in sustainable agriculture: a state of the art. In: Meena VS, Maurya BR, Verma JP,
Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture. Springer,
India, pp 21–29. doi:10.1007/978-81-322-2776-2_2
Jat LK, Singh YV, Meena SK, Meena SK, Parihar M, Jatav HS, Meena RK, Meena VS (2015)
Does integrated nutrient management enhance agricultural productivity? J Pure Appl Microbiol
9(2):1211–1221
Jha Y, Subramanian RB (2016) Regulation of plant physiology and antioxidant enzymes for alle-
viating salinity stress by potassium-mobilizing bacteria. In: Meena VS, Maurya BR, Verma JP,
Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture. Springer,
India, pp 149–162. doi:10.1007/978-81-322-2776-2_11
Joshi KK, Kumar V, Dubey RC, Maheshwari DK (2006) Effect of chemical fertilizer adaptive vari-
ants, Pseudomonas aeruginosa GRC2 and Azotobacter chroococcum AC1 on Macrophomina
phaseolina causing charcoal rot of Brassica juncea. Korean J Environ Agric 25:228–235
Kaushal M, Wani SP (2016) Plant-growth-promoting rhizobacteria: drought stress alleviators to
ameliorate crop production in drylands. Ann Microbiol 66(1):35–42
Kennedy AC (2005) Rhizosphere. In: Sylvia DM (ed) Principles and applications of soil microbi-
ology, 2nd edn. Pearson Education, Upper Saddle River, pp 242–262
Khan MS, Zaidi A, Wani PA, Oves M (2009) Role of plant growth promoting rhizobacteria in the
remediation of metal contaminated soils Environ. Chem Lett 7:1–19
Kloepper JW, Leong J, Teintze M, Schroth MN (1980) Pseudomonas siderophores: a mechanism
explaining disease suppressive soils. Curr Microbiol 4:317–320
Kloepper JW, Lifshitz R, Zablotowicz RM (1989) Free-living bacterial inocula for enhancing crop
productivity. Trends Biotechnol 7:39–44
Kloepper JW, Gutierrez-Estrada A, Mclnroy JA (2007) Photoperiod regulates elicitation of
growth promotion but not induced resistance by plant growth-promoting rhizobacteria. Can
J Microbiol 53(2):159–167
Kuan KB, Othman R, Rahim KA, Shamsuddin ZH (2016) Plant growth-promoting Rhizobacteria
inoculation to enhance vegetative growth, nitrogen fixation and nitrogen remobilisation of
maize under greenhouse conditions. PLOS ONE. doi:10.1371/journal.pone.0152478
3 PGPR: Heart of Soil and Their Role in Soil Fertility 63
Kumar A, Bahadur I, Maurya BR, Raghuwanshi R, Meena VS, Singh DK, Dixit J (2015)
Does a plant growth-promoting rhizobacteria enhance agricultural sustainability? J Pure
ApplMicrobiol 9:715–724
Kumar A, Meena R, Meena VS, Bisht JK, Pattanayak A (2016a) Towards the stress management
and environmental sustainability. J Clean Prod 137:821–822
Kumar A, Patel JS, Bahadur I, Meena VS (2016b) The molecular mechanisms of KSMs for
enhancement of crop production under organic farming. In: Meena VS, Maurya BR, Verma JP,
Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture. Springer,
India, pp 61–75. doi:10.1007/978-81-322-2776-2_5
Kumar A, Maurya BR, Raghuwanshi R, Meena VS, Islam MT (2017) Co-inoculation with
Enterobacter and Rhizobacteria on yield and nutrient uptake by wheat (Triticum aestivum L.) in
the alluvial soil under indo-gangetic plain of India. Journal of plant growth Regulation.:10.1007/
s00344-016-9663-5.
Lacey LA, Frutos R, Kaya HK, Vail P (2001) Insect pathogens as biological control agents: do they
have a future? Biol Control 21:230–248
Ladha JK, De Bruijn FJ, Malik KA (1997) Introduction: assessing opportunities for nitrogen fixa-
tion in rice – a frontier project. Plant Soil 194:1–10
Liu ZL, Sinclair JB (1992) Population dynamics of Bacillus megaterium strain B153-2-2 in the
rhizosphere of soybean. Phytopathology 82:1297–1301
Lloret L, Martinez-Romero E (2005) Evolution and phylogeny of rhizobia. Rev Latinoam
Microbiol 47:43–60
Lugtenberg BJJ, Bloemberg GV (2004) In: Ramos JL (ed) Pseudomonas Vol. 1. New York, Kluwer
academic/plenum publishers, p 403–430
Maheshwari DK, Dubey RC, Aeron A, Kumar B, Kumar S et al (2012) Integrated approach for
disease management and growth enhancement of Sesamum indicum L. utilizing Azotobacter
chroococcum TRA2 and chemical fertilizer. World J Microbiol Biotechnol 28:3015–3024
Mahmood S, Daur I, Al-Solaimani SG, Ahmad S, Madkour MH, Yasir M, Hirt H, Ali S, Ali Z
(2016a) Plant growth promoting Rhizobacteria and silicon synergistically enhance salinity tol-
erance of mung bean. Front Plant Sci 7:876
Mahmood K, Xu Z, El-Kereamy A, Casaretto JA, Rothstein SJ (2016b) The Arabidopsis tran-
scription factor ANAC032 represses anthocyanin biosynthesis in response to high sucrose and
oxidative and abiotic stresses. Front Plant Sci doi.org/10.3389/fpls.2016.01548
Martinez-Viveros O, Jorquera MA, Crowley DE, Gajardo G, Mora ML (2010) Mechanisms and
practical considerations involved in plant growth promotion by rhizobacteria. J Soil Sci Plant
Nutr 10:293–319
Masood S, Bano A (2016) Mechanism of potassium solubilization in the agricultural soils by
the help of soil microorganisms. In: Meena VS, Maurya BR, Verma JP, Meena RS (eds)
Potassium solubilizing microorganisms for sustainable agriculture. Springer, India, pp 137–
147. doi:10.1007/978-81-322-2776-2_10
Maurya BR, Meena VS, Meena OP (2014) Influence of Inceptisol and Alfisol’s potassium solubi-
lizing bacteria (KSB) isolates on release of K from waste mica. Vegetos 27:181–187
Mazurier S, Corberand T, Lemanceau P, Raaijmakers JM (2009) Phenazine antibiotics produced
by fluorescent pseudomonads contribute to natural soil suppressiveness to Fusarium wilt.
ISME J 3:977–991
Meena OP, Maurya BR, Meena VS (2013a) Influence of K-solubilizing bacteria on release of
potassium from waste mica. Agric Sust Dev 1:53–56
Meena VS, Maurya BR, Bohra JS, Verma R, Meena MD (2013b) Effect of concentrate manure
and nutrient levels on enzymatic activities and microbial population under submerged rice in
alluvium soil of Varanasi. Crop Res 45(1,2 & 3):6–12
Meena VS, Maurya BR, Verma R, Meena RS, Jatav GK, Meena SK, Meena SK (2013c) Soil
microbial population and selected enzyme activities as influenced by concentrate manure and
inorganic fertilizer in alluvium soil of Varanasi. Bioscan 8(3):931–935
Meena VS, Maurya BR, Bahadur I (2014a) Potassium solubilization by bacterial strain in waste
mica. Bang J Bot 43:235–237
64 I.P. Sharma et al.
Meena VS, Maurya BR, Verma JP (2014b) Does a rhizospheric microorganism enhance K+ avail-
ability in agricultural soils? Microbiol Res 169:337–347
Meena RS, Meena VS, Meena SK, Verma JP (2015a) The needs of healthy soils for a healthy
world. J Clean Prod 102:560–561
Meena RS, Meena VS, Meena SK, Verma JP (2015b) Towards the plant stress mitigate the agricul-
tural productivity: a book review. J Clean Prod 102:552–553
Meena VS, Maurya BR, Meena RS (2015c) Residual impact of wellgrow formulation and NPK on
growth and yield of wheat (Triticum aestivum L.) Bangladesh J Bot 44(1):143–146
Meena VS, Maurya BR, Verma JP, Aeron A, Kumar A, Kim K, Bajpai VK (2015d) Potassium
solubilizing rhizobacteria (KSR): Isolation, identification, and K-release dynamics from waste
mica. Ecol Eng 81:340–347
Meena VS, Meena SK, Verma JP, Meena RS, Ghosh BN (2015e) The needs of nutrient use efficiency
for sustainable agriculture. J Clean Prod 102:562–563. doi:10.1016/j.jclepro.2015.04.044
Meena VS, Verma JP, Meena SK (2015f) Towards the current scenario of nutrient use efficiency in
crop species. J Clean Prod 102:556–557. doi:10.1016/j.jclepro.2015.04.030
Meena MK, Gupta S, Datta S (2016a) Antifungal potential of PGPR, their growth promoting activ-
ity on seed germination and seedling growth of winter wheat and genetic variabilities among
bacterial isolates. Int J Cur Microbiol Appl Sci 5(1):235–243
Meena RK, Singh RK, Singh NP, Meena SK, Meena VS (2016b) Isolation of low temperature sur-
viving plant growth-promoting rhizobacteria (PGPR) from pea (Pisum sativum L.) and docu-
mentation of their plant growth promoting traits. Biocatalysis and agricultural. Biotechnology
4:806–811
Meena RS, Bohra JS, Singh SP, Meena VS, Verma JP, Verma SK, Sihag SK (2016c) Towards the
prime response of manure to enhance nutrient use efficiency and soil sustainability a current
need: a book review. J Clean Prod 112(1):1258–1260
Meena SK, Rakshit A, Meena VS (2016d) Effect of seed bio-priming and N doses under varied
soil type on nitrogen use efficiency (NUE) of wheat (Triticum aestivum L.) under greenhouse
conditions. Biocatal Agric Biotechnol 6:68–75
Meena VS, Bahadur I, Maurya BR, Kumar A, Meena RK, Meena SK, Verma JP (2016e) Potassium-
solubilizing microorganism in evergreen agriculture: an overview. In: Meena VS, Maurya BR,
Verma JP, Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture.
Springer, India, pp 1–20. doi:10.1007/978-81-322-2776-2_1
Meena VS, Meena SK, Bisht JK, Pattanayak A (2016f) Conservation agricultural practices in sus-
tainable food production. J Clean Prod 137:690–691
Parewa HP, Yadav J, Rakshit A, Meena VS, Karthikeyan N (2014) Plant growth promoting rhizo-
bacteria enhance growth and nutrient uptake of crops. Agric Sustain Dev 2(2):101–116
Pieterse CM, Leon-Reyes A, Van der Ent S, Van Wees SC (2009) Networking by small-molecule
hormones in plant immunity. Nat Chem Biol 5:308–316
Pinton R, Varanini Z, Nannipieri P (2001) The rhizosphere as a site of biochemical interactions
among soil components, plants and microorganisms Pinton R, Varanini Z, Nannipieri P The
Rhizosphere biochemistry and organic substances at the soil-plant interface, Marcel Dekker,
New York. 1–17
Prakash S, Verma JP (2016) Global perspective of potash for fertilizer production. In: Meena VS,
Maurya BR, Verma JP, Meena RS (eds) Potassium solubilizing microorganisms for sustainable
agriculture. Springer, India, pp 327–331. doi:10.1007/978-81-322-2776-2_23
Prasad R, Kumar M, Varma A (2015) Role of PGPR in soil fertility and plant health. In:
Egamberdieva D et al (eds) Plant-growth-promoting rhizobacteria (PGPR) and medicinal
plants, soil biology. Springer International Publishing, Switzerland, pp 247–260
Priyadharsini P, Muthukumar T (2016) Interactions between arbuscular mycorrhizal fungi and
potassium-solubilizing microorganisms on agricultural productivity. In: Meena VS, Maurya
BR, Verma JP, Meena RS (eds) Potassium solubilizing microorganisms for sustainable agricul-
ture. Springer, India, pp 111–125. doi:10.1007/978-81-322-2776-2_8
Raghavendra MP, Nayaka NC, Nuthan BR (2016) Role of rhizosphere microflora in potas-
sium solubilization. In: Meena VS, Maurya BR, Verma JP, Meena RS (eds) Potassium
3 PGPR: Heart of Soil and Their Role in Soil Fertility 65
Singh M, Dotaniya ML, Mishra A, Dotaniya CK, Regar KL, Lata M (2016) Role of biofertil-
izers in conservation agriculture. In: Bisht JK, Meena VS, Mishra PK, Pattanayak A (eds)
Conservation agriculture: an approach to combat climate change in Indian Himalaya. Springer,
Singapore, pp 113–134. doi:10.1007/978-981-10-2558-7_4
Sivasakhti S, Usharani G, Saranraj P (2014) Biocontrol potentiality of plant growth promoting
bacteria (PGPR)- Pseudomonas fluorescence and Bacillus subtilis: a review. Afr J Agri Res
9:1265–1277
Somers E, Vanderleijden J, Srinivasan M (2004) Rhizosphere bacterial signalling: a love parade
beneath our feet. Crit Rev Microbiol 30:205–240
Singh NP, Singh RK, Meena VS, Meena RK (2015) Can we use maize (Zea mays) rhizobacteria as
plant growth promoter. Vegetos 28(1):86–99. doi:10.5958/2229-4473.2015.00012.9
Spaepen S, Vanderleyden J (2011) Auxin and plant-microbe interactions. Cold Spring Harb
Perspect Biol 3(4):a001438. doi:10.1101/cshperspect.a001438
Sturz AV, Nowak J (2000) An endophytic community of rhizobacteria and the strategies requires to
create yield enhancing associations with crops. Appl Soil Ecol 15:183–190
Teotia P, Kumar V, Kumar M, Shrivastava N, Varma A (2016) Rhizosphere microbes: potassium
solubilization and crop productivity-present and future aspects. In: Meena VS, Maurya BR,
Verma JP, Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture.
Springer, India, pp 315–325. doi:10.1007/978-81-322-2776-2_22
Tisdale SL, Nelson WL (1975) Soil fertility and fertilizers, 3rd edn. Collier Macmillan Publication,
London
Trudgill DL, Blok VC (2001) Apomictic, polyphagous root-knot nematodes: exceptionally suc-
cessful and damaging biotrophic root pathogens. Annu Rev Phytopathol 39:53–77
Vacheron V, Desbrosses G, Bouffaud M, Touraine B, Moënne-Loccoz Y, Muller D, Legendre L,
Wisniewski-Dyé F, Prigent-Combaret C (2013) Plant growth-promoting rhizobacteria and root
system functioning. Front Plant Sci doi.org/10.3389/fpls.2013.00356
Vejan P, Abdullah R, Khadiran T, Ismail S, Boyce AN (2016) Role of plant growth promoting
Rhizobacteria in agricultural sustainability—a review. Molecules 21:1–17
Velazquez E, Silva LR, Ramírez-Bahena MH, Peix A (2016) Diversity of potassium-solubilizing
microorganisms and their interactions with plants. In: Meena VS, Maurya BR, Verma JP,
Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture. Springer,
India, pp 99–110. doi:10.1007/978-81-322-2776-2_7
Verma JP, Jaiswa DK, Meena VS, Meena RS (2015a) Current need of organic farming for enhanc-
ing sustainable agriculture. J Clean Prod 102:545–547
Verma JP, Jaiswal DK, Meena VS, Kumar A, Meena RS (2015b) Issues and challenges about sus-
tainable agriculture production for management of natural resources to sustain soil fertility and
health. J Clean Prod 107:793–794
Verma R, Maurya BR, Meena VS (2014) Integrated effect of bio-organics with chemical fertilizer
on growth, yield and quality of cabbage (Brassica oleracea var capitata). Indian J Agric Sci
84(8):914–919
Wang B, Seiler JR, Mei C (2015) Burkholderia phytofirmans strain PsJN advanced development
and altered leaf level physiology of switchgrass. Biomass Bioenergy 83:493–500
Wani PA, Khan MS, Zaidi A (2007) Synergistic effect of the inoculation with nitrogen-fixing and
phosphate-solubilizing rhizobacteria on performance of field-grown chickpea. J Plant Nutr Soil
Sci 170:283–287
Werner D (2004) Signalling in the rhizobia-legumes symbiosis. In: Varma A, Abbott L, Werner D,
Hampp R (eds) Plant surface microbiology. Springer, N.Y, pp 99–119
Yadav BK, Sidhu AS (2016) Dynamics of potassium and their bioavailability for plant nutrition.
In: Meena VS, Maurya BR, Verma JP, Meena RS (eds) Potassium solubilizing microorganisms
for sustainable agriculture. Springer, India, pp 187–201. doi:10.1007/978-81-322-2776-2_14
Yasin M, Munir I, Faisal M (2016) Can Bacillus spp. enhance K+ uptake in crop species. In:
Meena VS, Maurya BR, Verma JP, Meena RS (eds) Potassium solubilizing microorganisms
for sustainable agriculture. Springer, India, pp 163–170. doi:10.1007/978-81-322-2776-2_12
3 PGPR: Heart of Soil and Their Role in Soil Fertility 67
Youssef MMA, Eissa MFM (2014) Biofertilizers and their role in management of plant parasitic
nematodes. E J Biotechnol Pharm Res 5:1–6
Yu GY, Sinclair JB, Hartman GL, Bertagnolli BL (2002) Production of iturin A by Bacillus amylo-
liquefaciens suppressing Rhizoctonia solani. Soil Biol Biochem 34:955–963
Zahedi H (2016) Growth-promoting effect of potassium-solubilizing microorganisms on
some crop species. In: Meena VS, Maurya BR, Verma JP, Meena RS (eds) Potassium
solubilizing microorganisms for sustainable agriculture. Springer, India, pp 31–42.
doi:10.1007/978-81-322-2776-2_3
Zhang S, Moyne AL, Reddy MS, Kloepper JW (2002) The role of salicylic acid in induced sys-
temic resistance elicited by plant growth-promoting rhizobacteria against blue mold of tobacco.
Biol Control 25:288–296
Strength of Microbes in Nutrient Cycling:
A Key to Soil Health 4
Nisha Sahu, D. Vasu, Asha Sahu, Narayan Lal,
and S.K. Singh
Abstract
Nowadays, due to continuous degradation in soil quality, a healthy soil system is
the result of a complex network of physical, chemical, and biological soil quality
indicators. Healthy soils provide a balance between the needs of both farmers
and community. Soil organic matter (SOM) helps to sustain the soil health as
well as its quality, inactivate toxic compounds, suppress pathogens, and protect
environmental sustainability. It implies interactions among the soil’s internal and
external components for the sustainable food production system. The efficient
soil microbes play an important role, since they are responsible to drive various
biological transformations and different pools of carbon (C) and macro- and
micronutrients, which facilitate the subsequent establishment of soil-plant-
microbe interaction. The diversity of microbes in soil system is enormous. This
article emphasizes the role of microbes for soil health through the decomposition
of SOM present in soil system. Toward the global knowledge of soil microbial
dynamics, its function is increasing rapidly, but the knowledge of rhizospheric
complex is limited, despite of their importance in regulating soil-plant systems.
Keywords
Soil health • Microorganisms • Nutrient transformations • Sustainable
4.1 Introduction
The importance of soil is increasing at an alarming rate due to the rapidly growing
worldwide population as healthy soils will be less to meet the future food supply
due to growing pressure on land for urban expansion, biofuel production, and natu-
ral resource extraction (Meena et al. 2013a; Bahadur et al. 2014; Jha and Subramanian
2016; Kumar et al. 2016a, b). The soil develops by degeneration of rocks as well as
minerals, through biotic actions of microbes sustained by them (Bahadur et al.
2017; Verma et al. 2017b; Kumar et al. 2017a; Nath et al. 2017). It represents
dynamic ecosystems, making it appropriate to think about them in terms such as
health, vitality, and soil sustainability. Soils are the resources that provide humans
with ~90% of all the food we consume. Our one major challenge is to manage soils
in a sustainable fashion so that they will meet the needs of the next generation
(Maurya et al. 2014; Jat et al. 2015; Kumar et al. 2015; Ahmad et al. 2016; Meena
et al. 2015f, 2016a; Parewa et al. 2014; Dotaniya et al. 2016; Jaiswal et al. 2016).
Meanwhile, measurement of soil processes and its properties linked to these also
depends on the use and location of the soil. Previously, not only the physical and
chemical properties were given importance, but nowadays also the role of soil bio-
diversity in maintaining soil fertility and crop productivity (Singh 2015).
Soils are establishment of all terrestrial ecosystems and are considered home to
a vast diversity of rhizospheric macro- and microorganisms. They provide food or
nutrients to the macro-/microorganisms that live above and below the ground and
play significant role in buffering and filtering of soil system. Therefore, soils are
very important to existing societies (mainly human) as we depend on our day-to-day
work (Dominati et al. 2010; Prakash and Verma 2016; Meena et al. 2016b;
Priyadharsini and Muthukumar 2016; Kumar et al. 2017a; Bahadur et al. 2016b;
Das and Pradhan 2016; Dominguez-Nunez et al. 2016).
Soils support a diversity of life and conception is similar to human health; it is
not difficult to understand or recognize when the system is viewed as a whole. The
strength of soil comes from the life that exists within it – soil microorganisms, rang-
ing from genes and species to communities. It is an excellent indicator of soil health
(Nielsen and Winding 2002; Takoutsing et al. 2016). Variation in microbial popula-
tion and activities is considered as early signs of soil degradation and amelioration.
These efficient rhizospheric microorganisms are a vitally important component of
the soil habitat where they play significant roles in soil-plant systems functioning
through controlling nutrient cycling reactions essential for sustaining soil quality
and also contributing to the genesis and maintenance of soil structure (Meena et al.
2015a, b, e; Raghavendra et al. 2016; Zahedi 2016; Rawat et al. 2016; Yasin et al.
2016; Teotia et al. 2016).
There is an infinite abundance of microbes that exist within the soil system.
However, very little is known about the tiny creature which is responsible for numer-
ous processes occurring in soil and maintaining productivity in natural and man-
aged agroecosystems (Meena et al. 2016c, e; Saha et al. 2016a; Yadav and Sidhu
2016). Investigation on rhizospheric chemistry in relation to mainly nutrients and
microbes is difficult as we cannot see them, so they are out of sight and out of mind;
4 Strength of Microbes in Nutrient Cycling: A Key to Soil Health 71
most methods for studying these critters are rife with difficulties, and though we can
identify thousands of different species, we only really know what a small percent-
age of them are doing in the soil. Only ~1% of the microbes’ species have been
identified or cultured. In short, we have a lot left to learn by understanding what
microbes are doing, under what conditions they dominate, and how to sustain nutri-
ent cycling (Meena et al. 2014a, 2016d; Saha et al. 2016b; Verma et al. 2014, 2015b;
Bahadur et al. 2016a; Masood and Bano 2016).
Soil microbes have an enormous influence on soil-plant-microbe interactions
and play a significant role for maintaining soil quality. The most important of these
interactions is nutrient cycling. Under the healthy soil-plant system, soil biota regu-
lates the flow, root growth, and storage of nutrients. Even fertilization may pass
through efficient rhizospheric microbes before being utilized by the crop/plant
(Dubey et al. 2016; Verma et al. 2017a).
In the long-term soil fertility, the more we are supporting the next generation. A
healthy, fertile soil laid the groundwork for a strong and resilient food production
system. Nowadays, one of the major tasks is to manage and maintain soil in a sus-
tainable fashion. In the mid-1990s, the term soil health was introduced; several
scientists gave various definitions of soil health, and one of the most commonly
proposed definition is by Doran and Safley 1997 “The continued capacity of soil to
function as a vital living system, within ecosystem and land-use boundaries, to sus-
tain biological productivity, promote the quality of air and water environments, and
maintain plant, animal and human health.” This definition encompasses a time com-
ponent, reflecting the importance of continuous function over time and the dynamic
nature of soil-plant system (Sharma et al. 2016; Verma et al. 2015a; Meena et al.
2013c).
Soil is a finite and nonrenewable resource because regeneration of soil through
various weathering processes (physical, chemical, and biological) of underlying
rock requires geological time (Huber et al. 2001). The soil quality focuses on the
continued capacity or buffering capacity of soil to sustain plant growth promoting
(PGP) and is essential for the integrity of ecosystems to remain intact or to recover
from disturbances (Pankhurst et al. 1997). However, the deterioration of soil quality
is a major concern for human-animal-plant health (Singer and Ewing 2000).
Healthy soils are able to balance a range of functions to meet the needs of both
farmers and community. It functions to sustain soil biota and plant life, store and
cycle wastewater and nutrients, decompose SOM, inactivate toxic compounds, sup-
press pathogens, and protect soil and water quality. Meanwhile, healthy soil system
had a concept that implies that the soil functions as a balance of each other, and it
enhances the sustainability of production system (Meena et al. 2013b, 2015c;
Shrivastava et al. 2016; Velazquez et al. 2016; Singh et al. 2015). The degradation
in soil-water quality is a symptom of poor soil health (Takoutsing et al. 2016).
Healthy soils function to sustain biological productivity, store and cycle water and
72 N. Sahu et al.
The biological activity in soil is largely concentrated in the topsoil, where the depth
may vary from a few to ~120 cm. In the topsoil, the biological components (soil
microorganisms) occupy a tiny fraction (<0.5%) of the total soil volume and make
up <10% of SOM. They play a significant role in the cycling of nutrients and the
decomposition of organic residues (Pankhurst et al. 1997). The organic residues are,
in this way, converted to biomass or mineralized to CO2, H2O, and nutrients (Bloem
et al. 1997). These efficient microbes are further associated with the transformation
and degradation of waste materials and synthetic organic compounds (Meena et al.
2014b, 2015d; Singh et al. 2016; Sindhu et al. 2016).
The roles of these efficient rhizospheric microbes possess the ability to give an
integrated measure of soil sustainability, an aspect that cannot be obtained with
physical/chemical measures and/or analyses of diversity of higher organisms.
Microbes respond quickly to changes; hence, they rapidly adapt to environmental
conditions. This adaptation potentially allows microbial analyses to be discriminat-
ing in soil health assessment, and changes in microbial populations and activities
may therefore function as an excellent indicator of change in soil health (Kennedy
and Papendick 1995; Pankhurst and Hawke 1995; Singh 2015). Soil is a dynamic
habitat for a variety of life-forms and provides a mechanical support to plants from
which they extract nutrients. Soil organisms are constantly interacting with one
another; sometimes these interactions are mutually beneficial, symbiotic, and com-
petitive. It improves soil health because the “good” bacteria, fungi, nematodes, and
insects are able to outcompete the “bad” ones and also contribute greatly by break-
ing down organic materials to make nutrients available to plants. Microorganisms
act as an important link in the process of nutrient recycling. Thus, the same funda-
mental soil structure in different locations is found to support different biological
communities. The microflora and microfauna present in the soil are depicted in
Fig. 4.1. Soils are combinations of sand, silt, and clay particles and provide a diver-
sity of microhabitats which supports a wide range of organisms. These organisms
interact to modify the atmosphere between soil particles. The atmosphere within the
soil differs from the above ground due to the use of the available oxygen present in
Animals
Plants
Nematode
Rotifern
Diatoms
Autotrophic
Heterotrophic
Nitrogen Oxidising Carbon
Compounds Compounds
Bacteria requiring
Nitrogen fixing bacteria
combined nitrogen
4 Strength of Microbes in Nutrient Cycling: A Key to Soil Health
Aerobic Anaerobic
Aerobic Anaerobic
the soil by microbial and other metabolisms, and finally there is less oxygen beneath
the ground. Similarly, carbon dioxide is generated as a by-product of microbial
metabolism, and there is a higher concentration of carbon dioxide within soil than
above ground (Sarkar et al. 2017; Verma et al. 2017a; Kumar et al. 2017b).
Soils contain many aerobic and facultative organisms, and because of the micro-
bial manipulation of microenvironments, soils may harbor a large number of obli-
gate anaerobes. The distribution of soil microorganisms is presented in Table 4.1.
These rhizobacteria are the largest group of soil microbes, both in total number and
in diversity. Nowadays, the great advances of molecular biological techniques like
sensitive isotope studies which are yielding information on the metabolism of soil
microbes are being used for taxonomic studies (Singh 2015; Dubey 2016). The
cycling of nutrients is very important as without which, the continuation of life on
earth would be impossible, as essential nutrients would rapidly be taken up by
organisms and locked in a form that cannot be used by others. Soil is one part of the
ecosystem and supports all terrestrial life-forms; therefore, protection of soil is of
high priority, and a thorough understanding of soil enzyme activities is a critical
factor in assuring that soil remains healthy. A better understanding of soil enzymes
in maintaining the soil health will provide an opportunity for an integrated biologi-
cal assessment of soils. The response of microbes to environmental stress is rapid
compared to higher organisms, due to their high surface to volume ratio (Pankhurst
and Hawke 1995).
These efficient microbial communities can be considered as architects of soils
(Rajendhran and Gunasekaran 2008), and many ecosystem services that are linked
to ecosystems, including plant production, safeguarding of drinking water, or C
sequestration, are closely linked to microbial activities and their functional traits
(Torsvik and Ovreas 2002; Lombard et al. 2011). Studies on the development of
abiotic and biotic interactions are very complex (Ollivier et al. 2011). Microbes act
on a scale of μm3 and form biogeochemical interfaces with the soil matrix, shaping
their own environment (Totsche et al. 2010; Monier et al. 2011) Finally, most func-
tional traits, for example, the degradation of plant litter or the development of food
web structures and closed nutrient cycles, are not a result of a single organism but
of microbial communities which closely interact which each other (Aneja et al.
2006; Simon and Daniel 2011; Sharma et al. 2012).
4 Strength of Microbes in Nutrient Cycling: A Key to Soil Health 75
Soil organic matter acts as a storehouse for nitrogen, phosphorus, and sulfur. There
is very little inorganic nitrogen in soils and much of it is derived from organic forms.
The organic matter decomposition is largely an enzymatic process and facilitated by
extracellular hydrolytic enzymes produced by soil microorganisms. Soil enzymes
are involved in the biochemical processes during organic matter decomposition.
The important soil enzymes are amylase, arylsulfatase, cellulase, chitinase, dehy-
drogenase, phosphatase, and urease which are released from plants, animals, organic
compounds, and microorganisms and soils (Gupta et al. 1993; Ganeshamurthy et al.
1995). The rate of SOM decomposition is influenced by environmental factors such
as temperature, moisture content, and microbial diversity (Debnath and Hajra 1972).
In principle, the breakdown of organic matter involves four stages, viz., (a) break-
down of compounds that are easy to decompose – like sugars, starches, and pro-
teins; (b) breakdown of compounds that take several years to decompose like
cellulose (an insoluble carbohydrate found in plants) and lignins (a very compli-
cated structure that is part of wood); (c) breakdown of compounds that can take up
to ~10 years to decompose – like some waxes and the phenols; and (d) breakdown
of compounds that take thousands of years to decompose which include humus-like
substances which are the result of integration of compounds from breakdown prod-
ucts of plants and those generated by microorganisms (Chakroborty and Sen 1967).
Mineralization of organic matter is the biological process where organic compounds
in organic matter are chemically converted by the microorganisms in soil to simpler
organic compounds, other organic compounds, or mineralized nutrients (Fig. 4.2).
Therefore, incorporating organic matter into soils can change the amount of nitro-
gen (and other nutrients) available to plants, and those having a high C/N ratio will
probably cause some nitrogen deficiency in the crops/plants, at least in the short
term (Katherine et al. 2004; Ellert et al. 1997).
source of SOM:
litter, roots, dead organisms,
excrements of living organisms
CO2
immobilization
of ions in bodies
oxidative decomposition of organisms
humification: mineralisation:
synthesis of organic compounds organic compounds are converted
such as fulvic acids, humic acids to inorganic forms, release of minerals
The result of microbial mineralization is the release of energy, water, gases, etc.
on the one hand and formation of complex amorphous substance humus, called
humification, on the other hand. The rate of oxidation of biologically important ele-
ments such as nitrogen, carbon, sulfur, phosphorus, etc. during mineralization of
organic matter is of paramount importance to plants. Microorganisms play a vital
role to convert elements to usable forms to the plants for their use. Thus, the pro-
cesses of mineralization and humification run in nature hand in hand (Billings and
Ziegler 2005).
Soil enzymes are a group of enzymes whose usual inhabitants are the soil and are
continuously playing an important role in maintaining soil ecology, physical and
chemical properties, fertility, and soil health and responsible for overall process of
organic matter decomposition in the soil system (Dick et al. 1996). They are impor-
tant in catalyzing several vital reactions necessary for the life processes of microor-
ganisms in soils and the stabilization of soil structure, the decomposition of organic
wastes, organic matter formation, and nutrient cycling, hence playing an important
role in agriculture (Fig. 4.3). The enzyme levels in soil systems vary in amounts
primarily due to the fact that each soil type has different amounts of organic matter
4 Strength of Microbes in Nutrient Cycling: A Key to Soil Health 77
ORGANIC MATTER
Microbes
Simple sugars, Hemicelluloses, Tanins, lignin,
amino acid, celluloses, suberins,
amides, etc. proteins, etc. keratins,
chitin, etc.
Microbes Microbes
Microbes CO2, H2O,
CO2, H2O, Simpler compounds like NH3, etc.
NH3, etc. simple proteins, quinoid
Microbes substances, etc. Complex
substances like
Microbes quinines, Microbes
polyphenols,
etc.
Minerals
Minerals
Residue withstanding
microbial degradation Condensation
Humus
content, composition, and activity of its living organisms and intensity of biological
processes. In practice, the biochemical reactions are brought about largely through
the catalytic contribution of enzymes and variable substrates that serve as energy
sources for various microorganisms (Mandels and Reese 1999; Kertesz and Mirleau
2004; Liebich et al. 2007). The enzymes are the direct mediators for biological
catabolism of SOM and mineral components. Soil enzyme activities are closely
related to SOM and soil quality; the changes are much sooner than other parame-
ters, thus providing early indications of changes in soil sustainability (Dick et al.
1996).
These enzymes may include amylase, arylsulfatase, β-glucosidase, cellulase,
chitinase, dehydrogenase, phosphatase, protease, and urease released from plants,
animals, organic compounds, and microorganisms and soils (Ganeshamurthy et al.
1995; Xu et al. 2015). Factors affecting enzyme activity in soil are free enzymes
adsorbed or otherwise bound to soil organic and inorganic fraction, released into the
soil from lysed microbes through the action of bacteria, and accessible in dead non-
lysed cells and any metabolic activity of living cells as roots present in soil
system.
78 N. Sahu et al.
4.7 Amylase
4.8 Arylsulfatase
Arylsulfatases are typically widespread in nature as well as in soils and are respon-
sible for the hydrolysis of sulfate esters (Kertesz and Mirleau 2004). They are
secreted by rhizobacteria into the external environment as a response to sulfur limi-
tation, and its occurrence is often correlated with microbial biomass and rate of S
immobilization. This enzyme has a role in the hydrolysis of aromatic sulfate esters
(R–O–SO3) to phenols (R–OH) and sulfate or sulfate sulfur (SO42− or SO4–S). So
far, very little is known about specific microbial genera or species that play an
important role in the soil organosulfur circle in which arylsulfatases are the key
enzymes (Gupta et al. 1993; Ganeshamurthy et al. 1995).
4.9 β-Glucosidase
4.10 Cellulases
The cellulose is the most abundant organic compound in the biosphere, comprising
~50% of the biomass synthesized by photosynthetic fixation of CO2. Growth and
survival of microbes are important in most agricultural soils which depend on the
carbon source contained in the cellulose occurring in the soil (Srinivasulu and
Rangaswamy 2006). Cellulases are a group of enzymes that catalyze the degrada-
tion of cellulose; polysaccharides are built up of β-1,4-linked glucose units (Fig. 4.4).
4 Strength of Microbes in Nutrient Cycling: A Key to Soil Health 79
Fig. 4.4 The
decomposition of cellulose CH2OH CH2OH
to glucose O O
CELLULOSE
O
(linear β 1,4 glucan)
1,000 to 10,000
glucose units
Glucose
Unit n
CH2OH CH2OH
O O
CELLOBIOSE
O (can be transported
into cell)
β 1,4-glucosidase
CH2OH
O
GLUCOSE
an
ic
ae
rob
Glycolysis
rob
ae
ic
Many researchers reported that cellulases in soils are derived mainly from plant
debris incorporated into the soil, and a limited amount may also originate from
fungi and bacteria in soils. Since, cellulase enzymes play an important role in global
recycling of the most abundant polymer, cellulose, in nature, it would be of critical
importance to understand this enzyme better so that it may be used more regularly
as a predictive tool in our soil fertility programs (Saha et al. 2003; Muter et al.
2008).
4.11 Chitinase
Chitinase or chitinolytic enzymes are key enzymes responsible for the degradation
and hydrolysis of chitin (poly-β-1-4-(2-acetamido-2-deoxy)-D-glucoside). They are
also considered as the major structural component for many fungal cell walls that
use the hyperparasitism mechanisms against pest/pathogen attack. These biological
agents also reduce disease-producing agents by using other mechanisms such as
80 N. Sahu et al.
4.12 Dehydrogenase
4.13 Phosphatases
In soil ecosystems, these enzymes are believed to play critical roles in P cycles as
they are correlated to P stress and plant growth and are also good indicators of soil
fertility. For example, when there is a signal indicating P deficiency in the soil, acid
phosphatase secretion from plant roots is increased to enhance the solubilization
and remobilization of phosphate, thus influencing the ability of the plant to cope
with P-stressed conditions (Saha et al. 2003).
4.14 Proteases
Soil organisms (macro and micro) play a significant role in soil processes, including
nutrient cycling. These efficient processes are essential to agriculture and forestry
and for protecting the quality of water, air, and habitat. Soil enzymes act as a cata-
lyst to sustain soil health and its fertility. Enzymes have significant effects on soil
biology, environmental management, growth, and nutrient uptake in plants growing
in ecosystems. However, their activities may be influenced by unknown cultural
4 Strength of Microbes in Nutrient Cycling: A Key to Soil Health 81
Acknowledgments The authors are grateful to ICAR-National Bureau of Soil Survey and Land
Use Planning for technical support to carry out the work.
References
Ahmad M, Nadeem SM, Naveed M, Zahir ZA (2016) Potassium-solubilizing bacteria and their
application in agriculture. In: Meena VS, Maurya BR, Verma JP, Meena RS (eds) Potassium
solubilizing microorganisms for sustainable agriculture. Springer, India, pp 293–313.
doi:10.1007/978-81-322-2776-2_21
Aneja M, Sharma S et al (2006) Microbial degradation of beech litter – influence of soil type and
litter quality on the structure and function of microbial populations involved in the turnover
process. Microb Ecol 52:127–135
Bahadur I, Meena VS, Kumar S (2014) Importance and application of potassic biofertilizer in
Indian agriculture. Int Res J Biol Sci 3:80–85
Bahadur I, Maurya BR, Kumar A, Meena VS, Raghuwanshi R (2016a) Towards the soil sustain-
ability and potassium-solubilizing microorganisms. In: Meena VS, Maurya BR, Verma JP,
Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture. Springer,
India, pp 225–266. doi:10.1007/978-81-322-2776-2_18
Bahadur I, Maurya BR, Meena VS, Saha M, Kumar A, Aeron A (2016b) Mineral release dynamics
of tricalcium phosphate and waste muscovite by mineral-solubilizing rhizobacteria isolated
from indo-gangetic plain of India. Geomicrobiol J. doi:10.1080/01490451.2016.1219431
Bahadur I, Maurya BR, Meena VS, Saha M, Kumar A, Aeron A (2017) Mineral release dynamics
of tricalcium phosphate and waste muscovite by mineral-solubilizing rhizobacteria isolated
from indo-gangetic plain of India. Geomicrobiol J. doi:10.1080/01490451.2016.1219431
Billings SA, Ziegler SE (2005) Linking microbial activity and soil organic matter transformations
in forest soils under elevated CO2. Glob Chang Biol 11:203–212
Bloem J, Ruiter P et al (1997) Food webs and nutrient cycling in agro-ecosystems. In: van Elsas
JD, Trevors JT, Wellington EMH (eds) Modern soil microbiology. Marcel Dekker, New York,
pp 245–278
Chakroborty T, Sen S (1967) Mineralisation of carbon from added organic matter in the soil under
laboratory conditions. J Indian Soc Soil Sci 15:155–157
Das I, Pradhan M (2016) Potassium-solubilizing microorganisms and their role in enhancing
soil fertility and health. In: Meena VS, Maurya BR, Verma JP, Meena RS (eds) Potassium
solubilizing microorganisms for sustainable agriculture. Springer, India, pp 281–291.
doi:10.1007/978-81-322-2776-2_20
Debnath NC, Hajra JN (1972) Transformation of organic matter in soil in relation to mineralization
of carbon and nutrient availability. J Indian Soc Soil Sci 20(2):95–102
Dick RP, Breakwell DP, Turco RF (1996) Soil enzyme activities and biodiversity measurements as
integrative microbiological indicators. In: Doran JW, Jones AJ (eds) Methods of assessing soil
quality. Soil Science Society of America, Madison, pp 247–271
Dominati E, Patterson M et al (2010) A framework for classifying and quantifying natural capital
and ecosystem services of soils. Ecol Econ 69:1858–1868
82 N. Sahu et al.
Kumar A, Maurya BR, Raghuwanshi R, Meena VS, Islam MT (2017a) Co-inoculation with
enterobacter and rhizobacteria on yield and nutrient uptake by wheat (Triticum aestivum L.)
in the alluvial soil under indo-gangetic plain of India. J Plant Growth Regul. doi:10.1007/
s00344-016-9663-5
Kumar A, Meena VS, Maurya BR, Raghuwanshi R, Bisht JK, Pattanayak A (2017b) Towards the
biological nitrogen fixation and nitrogen management in legume under sustainable agriculture.
Appl Soil Ecol. doi:10.1016/j.apsoil.2017.05.013
Liebich J, Schloter M, Schaffer A, Vereecken H, Burauel P (2007) Degradation and humification
of maize straw in soil microcosms inoculated with simple and complex microbial communities.
Eur J Soil Sci 58:141–151
Lombard N, Prestat E et al (2011) Soil-specific limitations for access and analysis of soil microbial
communities by metagenomics. FEMS Microbiol Ecol 78:31–49
Mandels M, Reese ET (1999) Fungal cellulase and microbial degradation of cellulosic fabric. J Ind
Microbiol Biotechnol 22:225–240
Masood S, Bano A (2016) Mechanism of potassium solubilization in the agricultural soils by
the help of soil microorganisms. In: Meena VS, Maurya BR, Verma JP, Meena RS (eds)
Potassium solubilizing microorganisms for sustainable agriculture. Springer, India, pp 137–
147. doi:10.1007/978-81-322-2776-2_10
Maurya BR, Meena VS, Meena OP (2014) Influence of inceptisol and alfisol’s potassium solubiliz-
ing bacteria (KSB) isolates on release of K from waste mica. Vegetos 27:181–187
Meena OP, Maurya BR, Meena VS (2013a) Influence of K-solubilizing bacteria on release of
potassium from waste mica. Agric Sust Dev 1:53–56
Meena VS, Maurya BR, Bohra JS, Verma R, Meena MD (2013b) Effect of concentrate manure
and nutrient levels on enzymatic activities and microbial population under submerged rice in
alluvium soil of Varanasi. Crop Res 45(1,2 & 3):6–12
Meena VS, Maurya BR, Verma R, Meena RS, Jatav GK, Meena SK, Meena SK (2013c) Soil
microbial population and selected enzyme activities as influenced by concentrate manure and
inorganic fertilizer in alluvium soil of Varanasi. Bioscan 8(3):931–935
Meena VS, Maurya BR, Bahadur I (2014a) Potassium solubilization by bacterial strain in waste
mica. Bang J Bot 43:235–237
Meena VS, Maurya BR, Verma JP (2014b) Does a rhizospheric microorganism enhance K+ avail-
ability in agricultural soils? Microbiol Res 169:337–347
Meena RS, Meena VS, Meena SK, Verma JP (2015a) The needs of healthy soils for a healthy
world. J Clean Prod 102:560–561
Meena RS, Meena VS, Meena SK, Verma JP (2015b) Towards the plant stress mitigate the agricul-
tural productivity: a book review. J Clean Prod 102:552–553
Meena VS, Maurya BR, Verma JP, Aeron A, Kumar A, Kim K, Bajpai VK (2015c) Potassium
solubilizing rhizobacteria (KSR): isolation, identification, and K-release dynamics from waste
mica. Ecol Eng 81:340–347
Meena VS, Meena SK, Verma JP, Meena RS, Ghosh BN (2015d) The needs of nutrient use efficiency
for sustainable agriculture. J Clean Prod 102:562–563. doi:10.1016/j.jclepro.2015.04.044
Meena VS, Verma JP, Meena SK (2015e) Towards the current scenario of nutrient use efficiency in
crop species. J Clean Prod 102:556–557. doi:10.1016/j.jclepro.2015.04.030
Meena VS, Maurya BR, Meena RS (2015f) Residual impact of wellgrow formulation and NPK on
growth and yield of wheat (Triticum aestivum L.) Bangladesh J Bot 44(1):143–146
Meena RK, Singh RK, Singh NP, Meena SK, Meena VS (2016a) Isolation of low temperature sur-
viving plant growth-promoting rhizobacteria (PGPR) from pea (Pisum sativum L.) and docu-
mentation of their plant growth promoting traits. Biocatal Agric Biotechnol 4:806–811
Meena RS, Bohra JS, Singh SP, Meena VS, Verma JP, Verma SK, Sihag SK (2016b) Towards the
prime response of manure to enhance nutrient use efficiency and soil sustainability a current
need: a book review. J Clean Prod 112(1):1258–1260
Meena VS, Bahadur I, Maurya BR, Kumar A, Meena RK, Meena SK, Verma JP (2016c) Potassium-
solubilizing microorganism in evergreen agriculture: an overview. In: Meena VS, Maurya BR,
84 N. Sahu et al.
Verma JP, Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture.
Springer, India, pp 1–20. doi:10.1007/978-81-322-2776-2_1
Meena VS, Meena SK, Bisht JK, Pattanayak A (2016d) Conservation agricultural practices in
sustainable food production. J Clean Prod 137:690–691
Meena SK, Rakshit A, Meena VS (2016e) Effect of seed bio-priming and N doses under varied
soil type on nitrogen use efficiency (NUE) of wheat (Triticum aestivum L.) under greenhouse
conditions. Biocatal Agric Biotechnol 6:68–75
Monier JM, Demaneche S et al (2011) Metagenomic exploration of antibiotic resistance in soil.
Curr Opin Microbiol 14:229–235
Muter O, Versilovskis A, Scherbaka R, Grube M, Zarina D (2008) Effect of plant extract on the
degradation of nitroaromatic compounds by soil microorganisms. J Ind Microbiol Biotechnol
35:1539–1543
Nath D, Maurya BR, Meena VS (2017) Documentation of five potassium- and phosphorus-
solubilizing bacteria for their K and P-solubilization ability from various minerals. Biocatal
Agric Biotechnol 10:174–181
Nielsen MN, Winding A (2002) Microorganisms as indicators of soil health, NERI Technical
Report No. 388. National Environmental Research Institute, Ministry of the Environment,
Denmark
Ollivier J, Towe S et al (2011) Nitrogen turnover in soil and global change. FEMS Microbiol Ecol
78:3–16
Pankhurst CE, Hawke BG (1995) Evaluation of soil biological properties as potential bioindicators
of soil health. Aust J Exp Agric 35:1015–1028
Pankhurst CE, Doube BM et al (1997) Biological indicators of soil health: synthesis. In: Pankhurst
CE, Doube BM, Gupta VVSR (eds) Biological indicators of soil health. CAB International,
Wallingford, pp 419–435
Parewa HP, Yadav J, Rakshit A, Meena VS, Karthikeyan N (2014) Plant growth promoting rhizo-
bacteria enhance growth and nutrient uptake of crops. Agric Sustain Dev 2(2):101–116
Parr JF, Papendick RI et al (1992) Soil quality: attributes and relationship to alternative and sus-
tainable agriculture. Am J Altern Agric 7:5–11
Prakash S, Verma JP (2016) Global perspective of potash for fertilizer production. In: Meena VS,
Maurya BR, Verma JP, Meena RS (eds) Potassium solubilizing microorganisms for sustainable
agriculture. Springer, India, pp 327–331. doi:10.1007/978-81-322-2776-2_23
Priyadharsini P, Muthukumar T (2016) Interactions between arbuscular mycorrhizal fungi and
potassium-solubilizing microorganisms on agricultural productivity. In: Meena VS, Maurya
BR, Verma JP, Meena RS (eds) Potassium solubilizing microorganisms for sustainable agricul-
ture. Springer, India, pp 111–125. doi:10.1007/978-81-322-2776-2_8
Raghavendra MP, Nayaka NC, Nuthan BR (2016) Role of rhizosphere microflora in potas-
sium solubilization. In: Meena VS, Maurya BR, Verma JP, Meena RS (eds) Potassium
solubilizing microorganisms for sustainable agriculture. Springer, India, pp 43–59.
doi:10.1007/978-81-322-2776-2_4
Rajendhran J, Gunasekaran P (2008) Strategies for accessing soil metagenome for desired applica-
tions. Biotechnol Adv 26:576–590
Rawat J, Sanwal P, Saxena J (2016) Potassium and its role in sustainable agriculture. In: Meena
VS, Maurya BR, Verma JP, Meena RS (eds) Potassium solubilizing microorganisms for sus-
tainable agriculture. Springer, India, pp 235–253. doi:10.1007/978-81-322-2776-2_17
Saha H, Mandal S, Hajra JN (2003) Effect of phosphate rock and pyrite on the decomposition and
enrichment of rice straw inoculated with cellulose decomposing fungi and cowdung slurry.
Indian J Agric 47(3–4):265–272
Saha M, Maurya BR, Bahadur I, Kumar A, Meena VS (2016a) Can potassium-solubilising bac-
teria mitigate the potassium problems in India? In: Meena VS, Maurya BR, Verma JP, Meena
RS (eds) Potassium solubilizing microorganisms for sustainable agriculture. Springer, India,
pp 127–136. doi:10.1007/978-81-322-2776-2_9
4 Strength of Microbes in Nutrient Cycling: A Key to Soil Health 85
Saha M, Maurya BR, Meena VS, Bahadur I, Kumar A (2016b) Identification and characterization
of potassium solubilizing bacteria (KSB) from Indo-Gangetic Plains of India. Biocatal Agric
Biotechnol 7:202–209
Sarkar D, Meena VS, Haldar A, Rakshit R (2017) Site-specific nutrient management (SSNM):
a unique approach towards maintaining soil health. Adapt Soil Manag: Theory Pract:69–88.
doi:10.1007/978-981-10-3638-5_3
Sharma S, Mehta R et al (2012) Improved protocol for the extraction of bacterial mRNA from
soils. J Microbiol Methods 91:62–64
Sharma A, Shankhdhar D, Shankhdhar SC (2016) Potassium-solubilizing microorganisms:
mechanism and their role in potassium solubilization and uptake. In: Meena VS, Maurya BR,
Verma JP, Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture.
Springer, India, pp 203–219. doi:10.1007/978-81-322-2776-2_15
Shrivastava M, Srivastava PC, D’Souza SF (2016) KSM soil diversity and mineral solubilization,
in relation to crop production and molecular mechanism. In: Meena VS, Maurya BR, Verma JP,
Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture. Springer,
India, pp 221–234. doi:10.1007/978-81-322-2776-2_16
Simon C, Daniel R (2011) Metagenomic analyses: past and future trends. Appl Environ Microbiol
77:1153–1161
Sindhu SS, Parmar P, Phour M, Sehrawat A (2016) Potassium-solubilizing microorganisms
(KSMs) and its effect on plant growth improvement. In: Meena VS, Maurya BR, Verma JP,
Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture. Springer,
India, pp 171–185. doi:10.1007/978-81-322-2776-2_13
Singer MJ, Ewing S (2000) Soil quality. In: Sumner ME (ed) Handbook of soil science. CRC
Press, Boca Raton, pp G271–G298
Singh JS (2015) Plant-microbe interactions: a viable tool for agricultural sustainability. Appl Soil
Ecol 92:45–46
Singh NP, Singh RK, Meena VS, Meena RK (2015) Can we use maize (Zea mays) rhizobacteria as
plant growth promoter? Vegetos 28(1):86–99. doi:10.5958/2229-4473.2015.00012.9
Singh M, Dotaniya ML, Mishra A, Dotaniya CK, Regar KL, Lata M (2016) Role of biofertil-
izers in conservation agriculture. In: Bisht JK, Meena VS, Mishra PK, Pattanayak A (eds)
Conservation agriculture: an approach to combat climate change in Indian Himalaya. Springer,
Singapore, pp 113–134. doi:10.1007/978-981-10-2558-7_4
Srinivasulu M, Rangaswamy V (2006) Activities of invertase and cellulase as influenced by the
application of tridemorph and captan to groundnut (Arachis hypogaea) soil. Afr J Biotechnol
5:175–180
Takoutsing B, Weber J et al (2016) Assessment of soil health indicators for sustainable production
of maize in small holder farming systems in the highlands of Cameroon. Geoderma 276:64–73
Teotia P, Kumar V, Kumar M, Shrivastava N, Varma A (2016) Rhizosphere microbes: potassium
solubilization and crop productivity-present and future aspects. In: Meena VS, Maurya BR,
Verma JP, Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture.
Springer, India, pp 315–325. doi:10.1007/978-81-322-2776-2_22
Torsvik V, Ovreas L (2002) Microbial diversity and function in soil: from genes to ecosystems.
Curr Opin Microbiol 5(3):240–245
Totsche KU, Rennert T, Gerzabek MH, Kogel-Knabner I, Smalla K, Spiteller M, Vogel HJ (2010)
Biogeochemical interfaces in soil: the interdisciplinary challenge for soil science. J Plant Nutr
Soil Sci 173:88–99
Vasilchenko LG, Karapetyan KN, Yachkova SN, Zernova ES, Rabinovich ML (2004) Degradation
of a lignin–carbohydrate substrate by soil fungi producing laccase and cellobiose dehydroge-
nase. Appl Biochem Microbiol 40(1):44–48
Velazquez E, Silva LR, Ramírez-Bahena MH, Peix A (2016) Diversity of potassium-solubilizing
microorganisms and their interactions with plants. In: Meena VS, Maurya BR, Verma JP,
Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture. Springer,
India, pp 99–110. doi:10.1007/978-81-322-2776-2_7
86 N. Sahu et al.
Verma R, Maurya BR, Meena VS (2014) Integrated effect of bio-organics with chemical fertilizer
on growth, yield and quality of cabbage (Brassica oleracea var capitata). Indian J Agric Sci
84(8):914–919
Verma JP, Jaiswa DK, Meena VS, Meena RS (2015a) Current need of organic farming for enhanc-
ing sustainable agriculture. J Clean Prod 102:545–547
Verma JP, Jaiswal DK, Meena VS, Kumar A, Meena RS (2015b) Issues and challenges about sus-
tainable agriculture production for management of natural resources to sustain soil fertility and
health. J Clean Prod 107:793–794
Verma R, Maurya BR, Meena VS, Dotaniya ML, Deewan P (2017a) Microbial dynamics as
influenced by bio-organics and mineral fertilizer in alluvium soil of Varanasi. India Int J Curr
Microbiol App Sci 6(2):1516–1524
Verma R, Maurya BR, Meena VS, Dotaniya ML, Deewan P, Jajoria M (2017b) Enhancing pro-
duction potential of cabbage and improves soil fertility status of Indo-Gangetic Plain through
application of bio-organics and mineral fertilizer. Int J Curr Microbiol App Sci 6(3):301–309
Xu Z, Yu G et al (2015) The variations in soil microbial communities, enzyme activities and their
relationship with soil organic matter decomposition along the northeren slope of Changbai
mountain. Appl Soil Ecol 86:19–29
Yadav BK, Sidhu AS (2016) Dynamics of potassium and their bioavailability for plant nutrition.
In: Meena VS, Maurya BR, Verma JP, Meena RS (eds) Potassium solubilizing microorganisms
for sustainable agriculture. Springer, India, pp 187–201. doi:10.1007/978-81-322-2776-2_14
Yasin M, Munir I, Faisal M (2016) Can Bacillus spp. enhance K+ uptake in crop species. In:
Meena VS, Maurya BR, Verma JP, Meena RS (eds) Potassium solubilizing microorganisms
for sustainable agriculture. Springer, India, pp 163–170. doi:10.1007/978-81-322-2776-2_12
Zahedi H (2016) Growth-promoting effect of potassium-solubilizing microorganisms on
some crop species. In: Meena VS, Maurya BR, Verma JP, Meena RS (eds) Potassium
solubilizing microorganisms for sustainable agriculture. Springer, India, pp 31–42.
doi:10.1007/978-81-322-2776-2_3
Quorum Sensing in Plant Microbe
Interaction 5
Raghavendra M.P.
Abstract
The coordinated behaviors, at the population level, occur in bacteria due to com-
munication between the cells, which is defined as quorum sensing (QS). These
communications help the bacteria to respond in terms of extracellular factors
which are involved in scavenging for nutrients; biosynthesis of exopolysaccha-
rides, extracellular hydrolytic enzymes, siderophores, antibiotics, pigments,
hypersensitive reaction; aiding motility; and providing scaffolding for biofilms
to grow as well as those involved in conjugation and epiphytic fitness. Even in
parasitic species, these signals help in regulation of bacterial growth, virulence,
and damage caused to hosts. QS signal molecules are regulated by autoinduction
and positive feedback mechanism at high cell densities. A detailed mechanism
involved in QS is discussed in this chapter with special reference to rhizosphere
microflora. The rhizosphere is a digestive system of the plants wherein several
microorganisms influenced by root exudates in turn favor its growth. In the
future, understanding QS related to plant growth-promoting bacteria and its
interspecific bacterial communication may offer good opportunities to manipulate
bacterial community structure in the rhizosphere for the benefit of agriculture.
Keywords
Quorum sensing • Rhizosphere microflora • Plant growth
R. M.P. (*)
Department of Microbiology, Maharani’s Science College for Women,
JLB Road, Mysuru 570005, Karnataka, India
e-mail: mpraghavendra@gmail.com
5.1 Introduction
Revitalization of the native soil systems for improved crop yield was emphasized in
many literatures and even the study conducted by United Nations’ Economic and
Social Commission for Asia and the Pacific on “Sustainable agriculture and food
security in Asia and the Pacific” emphasizes the same. There is a need of carrying
out these processes using various eco-friendly biological amendments especially
microorganisms. These organisms colonized in the rhizosphere share symbiotic and
asymbiotic relationship with plants benefiting its growth directly or indirectly
(Meena et al. 2013a; Bahadur et al. 2014; Kumar et al. 2016b).
Rhizosphere microflora along with other eukaryotic cell types is known to
produce specific small signaling molecules through which they are able to sense
their concentration in the environment. This behavior of accumulating signal
molecules in response to increase in population density of the specific organism is
named as quorum sensing (Fuqua and Greenberg 2002). The coordinated expres-
sion of the specific target genes of the entire population occurs when these signals
reach the threshold. These interactions help these organisms to occupy particular
habitat adapting to environmental conditions and resource levels and even to gener-
ate a coordinated protective response against several adverse conditions. These
coordinated responses are generally induced by a group of chemical signaling and
are referred to as quorum sensing (QS). QS signals produced by rhizosphere micro-
flora are significantly higher compared to other organisms isolated from bulk soil. It
is also evident from the work carried out on the QS signals with distinct chemical
structures produced by Gram-negative rhizosphere bacteria and their potential to
regulate a wide array of genes in the population (Hartmann et al. 2014; Maurya
et al. 2014; Jat et al. 2015; Kumar et al. 2015, 2016a; Jha and Subramanian 2016).
The rhizosphere is defined as the soil around the roots which is under the influence
of root exudates, whereas rhizoplane comprises of the microorganisms adhered
onto the root surface. Roots are reported to secrete exudates with a wide array of
chemicals involved in regulation of both beneficial and pathogenic microorganisms
in the rhizosphere. The variation in these chemicals helps in stabilizing the equi-
librium of beneficial microbes in terms of its number and by which evading the
continuous attack by soilborne pathogenic bacteria (Walker et al. 2003; Bais et al.
2003). The cross talk between the plant roots and beneficial microbes is playing a
vital role in growth and development of the entire plant kingdom in general and
agricultural crops in particular (Manefield and Turner 2002; Kent and Triplett 2002;
Sharma et al. 2003). The release of chemicals involved in trans-specific communi-
cation from roots can occur passively upon decay and may export signals actively to
the extracellular environment.
Ten to 40% of the photosynthetic carbon assimilation is released by the plants as
root exudates, and it is a mixture of sugars, amino acids, organic acids, sugar
5 Quorum Sensing in Plant Microbe Interaction 89
alcohols, and secondary metabolites (Bais et al. 2006). Due to its regular addition to
soil than the senescent plant debris, their role in regulating the microbial activity
through nutrient bioavailability and regulating phytotoxic elements is vital (Singh
and Mukerji 2006; Neumann 2007). Thus, microbial activity including cell division,
sporulation, and synthesis of secondary metabolites and even its number is very
high in the vicinity of growing roots than the non-rhizosphere soil. It is observed
that there is 12- to 25-fold difference in bacterial and fungal population between
these soils (Kennedy 2005; Parewa et al. 2014; Ahmad et al. 2016; Jaiswal et al.
2016; Meena et al. 2016a).
The rhizosphere is a hotspot of several communications involving a wide range
of microorganisms with diverse physiological importance (Bahadur et al. 2017;
Verma et al. 2017b; Kumar et al. 2017a; Nath et al. 2017). Among these communi-
cations, quorum sensing and quorum quenching are attracting scientific community
for its beneficial exploitation in plant growth regulations ultimately leading to yield
enhancement. Gram-negative bacteria use homoserine lactones (LuxR/LuxI) as
communication signals (Swift et al. 1999; Schauder and Bassler 2001), whereas
Gram-positive bacteria use oligopeptides in quorum sensing. The mechanisms of
quorum sensing are discussed in detail later in this chapter.
During quorum sensing, it is important for the organisms to differentiate between
species-specific signaling and signaling associated with interspecies behavior mod-
ulations (Bassler 1999; Federle and Bassler 2003; Waters and Bassler 2006). Hence,
they need to have competence in communication, which enables them to coordinate
behaviors related to species specifically and also to coordinate with other diverse
group of species (Hughes and Sperandio 2008). Mycorrhizal fungi stand as a best
example for this; these fungi support the growth of bacteria by releasing few nutri-
ents, and in turn soil bacteria with its wide array of enzymes degrade the complex
soil organic nutrients and make it easily available for the fungi (Bonfante 2003;
Bonfante and Anca 2009). It also supports the plant growth by extending its hyphae
to the areas where plant roots are not able to reach; due to this extension, the plant
gets sufficient nutrients supplied by both roots and fungi compared to uninfected
roots. Volumes of information are available in these interactions, and it is also docu-
mented that if mycorrhizal associations were not available, several plants would
have become extinct by this time (Meena et al. 2015f, 2016b; Dominguez-Nunez
et al. 2016; Dotaniya et al. 2016; Prakash and Verma 2016).
Smaller groups of the similar bacteria on the other hand use quorum sensing to
develop dialects required to occupy specific niche in the ecosystem. It also helps in
identifying the nonself species through species-specific and group-specific quorum
sensing. While facing the competition for specific niche, the complex coordination
among the species avoids the confusion and enables them to colonize the specific
habitat with specialized characteristics acquired through such interactions (Taga
and Bassler 2003). The law of limiting factor supported by adaptive radiation of
the species is favored by coexistence of bacterial life with suitable interactions to
scavenge the limiting factor required to colonize the specific habitat and suitably
favored in competition with other groups of organisms (Sarkar et al. 2017; Verma
et al. 2017a).
90 R. M.P.
(a) (b)
signal molecules
nutrient
concentration mass transfer
individual sensing
collective sensing
individual response
density
collective response
mass transfer
public goods
Fig. 5.1 Individual sensing versus collective sensing and responses; (a) individual sensing,
(b) collective sensing (Popat et al. 2015)
Generally, quorum sensing in bacteria falls into three classes: the first is, as men-
tioned earlier, AHL-dependent LuxI/LuxR-type QS observed in Gram-negative
bacteria, the second is the small peptide-mediated QS observed in Gram-positive
bacteria, and the third observed in both these bacteria is luxS-encoded autoinducer
2 (AI-2) QS. These signal molecules are operating with precise sensing and regula-
tory network (Dunny and Leonard 1997; Miller and Bassler 2001; Schauder and
Bassler 2001; Federle and Bassler 2003; Waters and Bassler 2005; Papenfort and
Bassler 2016).
The detailed QS mechanism associated with Gram-negative bacteria mediated
by AHL is available. AHL is freely diffusible and its concentration varies accord-
ingly in proportion to cell density. The specific target genes are activated when
LuxR-like protein recognizes AHL and binds to specific promoters on DNA, which
in turn positively regulates the genes leading to the production of proteins. LuxI-like
enzymes are reported to be responsible for production of specific AHL in associa-
tion with acyl-ACP of the fatty acid metabolism (Miller and Bassler 2001; Fuqua
and Greenberg 2002; Taga and Bassler 2003; Waters and Bassler 2005).
92 R. M.P.
Proteobacteria are reported as major colonizers of the rhizosphere, and hence the
QS signals N-acyl-homoserine lactones (AHLs) produced by them are significantly
high in the rhizosphere. More than 200 species of proteobacteria are known to
produce about 40 different acyl-HSL signals with different specificities and are pro-
duced by the LuxI family of signal synthases which binds to highly specific recep-
tors in the LuxR family that act as transcription factors. Most of the rhizosphere
microflora is reported to exploit this coevolved signal and receptor circuit for its
effective communication in the soil.
The major group observed in the rhizosphere is actinobacteria; in this group, QS
is dependent on the production of autoregulatory factors of different chemical
classes, mainly the butyrolactone class and the nucleotide-like B-factor class. The
5 Quorum Sensing in Plant Microbe Interaction 93
first class includes the A-factor of Streptomyces griseus (Yamada and Nihira 1998)
and similar compounds found in other actinobacteria (Yamada et al. 1987;
Kawabuchi et al. 1997). The nucleotide-like B-factor class has been isolated from
Amycolatopsis (Nocardia) mediterranei (Kawaguchi et al. 1988). In actinobacteria,
research efforts on QS have mostly focused on morphological differentiation and
secondary metabolism. The role of QS during the interaction between actinobacte-
ria and plants has not been examined in detail (Simao-Beaunoir et al. 2009).
Cell-to-cell signaling regulates the expression of the rhlAB operon responsible for
production of biosurfactants (Ochsner et al. 1994; Ochsner and Reiser 1995; Pearson
et al. 1997; Pesci et al. 1997). RhlI, N-butyrylhomoserine lactone autoinducer syn-
thase gene, and transcriptional activator encoding rhlR that are the major QS system
and rhamnosyltransferase encoded by rhlC which is coordinately regulated along
with rhlAB are responsible for biosurfactant production in microorganisms (Rahim
et al. 2001). These systems are under the influence of nutritional factor and QS sig-
nals (Guerra-Santos et al. 1986). They also came out with interesting observation
that the nutritional conditions supersede cell-to-cell communication and hence cor-
relate more positively with upregulation of quorum sensing-controlled genes such
as rhlAB. A similar conclusion was also expressed by Bollinger et al. (2001) related
to sodA encoding manganese-cofactored superoxide dismutase. These reports high-
light the need of understanding the role of nutritional factors and the QS signals in
cell density-based gene regulation in microorganisms.
Through social traits bacteria get several benefits such as coordinated population
behavior (Vibrio fischeri, Ps. aeruginosa, and Staph. aureus), biofilm formation to
get protection from adverse environmental conditions, nutrient and niche protection
in nocules (Rhizobium sp.), enhanced colonization and growth in specialized niches
(siderophores production for iron acquisition in bacteria), autolysis to provide nutri-
ents and DNA for biofilm development (Ps. aeruginosa), coordinated movement
toward nutrient source (Yersinia sp., Myxococcus xanthus, Ps. aeruginosa), antibi-
otic resistance through production of extracellular enzymes to break down antimi-
crobials (E. coli and Klebsiella spp.), and also immune modulation to facilitate
survival within the host (Ps. aeruginosa, Porphyromonas gingivalis, Helicobacter
pylori) (Diggle et al. 2007).
Microorganisms being small are suitable models to understand the basic questions
related to ecology and evolution of social interactions (Zahavi and Ralt 1984; Crespi
2001; Velicer 2003). In recent times, the beneficial traits acquired by these organ-
isms in high-density population are developing a new insight into their community
5 Quorum Sensing in Plant Microbe Interaction 95
Table 5.1 Biofilm formation by beneficial and pathogenic bacteria associated with plant roots
Bacteria Nature of association Plant name References
Acinetobacter PGPR Duckweed Yamaga et al. (2010)
calcoaceticus P23
Azospirillum brasilense PGPR Wheat Kim et al. (2005) and
Sheludko et al. (2010)
Azorhizobium caulinodans PGPR Rice Van Nieuwenhove
et al. (2004)
Azotobacter chroococcum PGPR Cotton, wheat Kumar et al. (2007)
Bacillus amyloliquefaciens PGPR, biocontrol Arabidopsis Fan et al. (2011)
S499 thaliana Nihorimbere et al.
(2012)
Bacillus cereus Under conditions of Wild barley Trivedi et al. (2011)
stress by salt, heat, or
desiccation
Bacillus pumilus Under conditions of Wild barley Trivedi et al. (2011)
stress by salt, heat, or
desiccation
Bacillus polymyxa PGPR Cucumber Yang et al. 2004
Nihorimbere et al.
(2012)
Bacillus subtilis Biocontrol Arabidopsis Ellis and Cooper
thaliana (2010) and
Beauregard et al.
(2013)
Bacillus megaterium C4 Nitrogen fixation, Maize, rice Liu et al. (2006)
PGPR
Burkholderia cepacia Lu Biocontrol Mulberry Ji et al. (2010)
10-1
Enterobacter agglomerans Biocontrol Cotton Chernin et al. (1995)
Enterobacter cloacae PGPR Rice Shankar et al. (2011)
Klebsiella pneumoniae Beneficial Wheat Dong et al. (2004)
and Liu et al. (2011)
Microsphaeropsis sp. Biocontrol Onion Carisse et al. (2001)
Micrococcus sp. NII-0909 PGPR Cowpea Dastager et al. (2010)
Paenibacillus lentimorbus Heavy metal Chickpea Khan et al. (2012)
tolerance
Paenibacillus polymyxa Biocontrol Peanut Haggag and Timmusk
(2008)
Pantoea agglomerans PGPR Chickpea, wheat Chauhan and Nautiyal
(2010)
Pseudomonas Biocontrol Wheat Sigler et al. 2001
aureofaciens
Pseudomonas Biocontrol Arabidopsis Lalaouna et al. (2012)
brassicacearum thaliana
Pseudomonas Biocontrol Wheat Chin-A-Woeng et al.
chlororaphis (2000) and Shen et al.
(2012)
(continued)
96 R. M.P.
Table 5.1 (continued)
Bacteria Nature of association Plant name References
Pseudomonas fluorescens Biocontrol Crop plant Silby and Levy
(2004) and Barahona
et al. (2010)
Pseudomonas putida Drought tolerance, Maize, Sandhya et al. (2009),
bioremediation sunflower, A. Matilla et al. (2011),
thaliana and Jakovleva et al.
(2012)
Pseudomonas aurantiaca PGPR Maize, wheat Rosas et al. (2009)
SR1
Rhizobium alamii Heavy metal Arabidopsis Schue et al. (2011)
tolerance thaliana,
rapeseed
Rhizobium leguminosarum Nitrogen fixation, Various legumes Fujishige et al.
pv. viciae 3841 PGPR, drought (2006), Williams et al.
tolerance (2008), and Janczarek
and Skorupska (2011)
Rhizobium leguminosarum Beneficial Rice Janczarek and
Skorupska (2011)
Rhizobium sp. NGR234 Nitrogen fixation, Legumes Kysciak et al. (2011)
PGPR (cowpea)
Rhizobium Symbiosis Legumes Fujishige et al. (2006)
and Robledo et al.
(2012)
Sinorhizobium Symbiosis Legumes Fujishige et al.
(2006), Khan et al.
(2012), and Amaya-
Gomez et al. (2015)
Stenotrophomonas Biocontrol, PGPR Crop plant Ryan et al. (2008)
maltophilia
Shewanella putrefaciens Microbial mediated Huang et al. (2011)
CN-32 geochemistry
Cyanobacteria sp. PGPR, biocontrol Prasanna et al. (2011)
signaling pathways are well documented and described (Loh et al. 2002; Newton
and Fray 2004; Nihorimbere et al. 2011; Podile et al. 2014).
The link between soil signaling and nitrogen cycling is also investigated by
DeAngelis et al. (2008). They reported that many alpha-proteobacteria were newly
found with QS-controlled extracellular enzyme activity, and even cell division,
symbiotic plasmid transfer, gene expression in the rhizosphere, symbiosome
development and nitrogen fixation, and nodule number in Rhizobium bacteria are
regulated by QS. On the contrary, QS is also reported to play an important role in
expression of genes associated with virulence factors (Meena et al. 2013c, 2016c;
Saha et al. 2016a; Yadav and Sidhu 2016; Singh et al. 2015; Bahadur et al. 2016a).
This supports an idea that developing QS mutants may be the next line of therapeu-
tic target for disease management.
There is a positive relationship between population growth rate and population
density in some species, which is first introduced by Allee et al. (1949) and hence
known as the Allee effect. He suggested that there exist threshold densities for some
species under some conditions, below which populations decline and thereby
increase the likelihood of extinction and above which population growth rate
increases as a function of density (Courchamp et al. 1999; Stephens and Sutherland
1999). This effect nowadays received renewed interest in conservation of plant and
animal diversity, and it also suits very well into the concepts of QS in microorgan-
isms (Darch et al. 2012; Meena et al. 2014b, 2015d, 2016d; Saha et al. 2016b;
Verma et al. 2015b; Singh et al. 2016).
Soil is the central organizer of the terrestrial ecosystem, and its physical, chemical,
and biological processes have enormous impacts on ecosystem productivity, ser-
vices, integrity, and human welfare. It is considered as a natural media for growth of
several microorganisms associated with plant growth. The overall contribution of
soil to the growth of plants is defined as the soil fertility. Several microorganisms
inhabit the soil plant interface and create microenvironment which is physiologi-
cally dynamic supporting the biogeochemical cycle. These microorganisms in turn
develop interactions which is an additional benefit for the plant growth (Verma et al.
2014; Meena et al. 2013b, 2014a, 2015c; Sharma et al. 2016; Verma et al. 2015a;
Shrivastava et al. 2016; Velazquez et al. 2016; Sindhu et al. 2016). To sense environ-
mental conditions and to change patterns of gene regulation accordingly, soil bacte-
ria have evolved sophisticated signal transduction mechanisms. Soil microorganisms
in general and rhizosphere microflora in particular are considered as treasure houses
of the soil defining its fertility and plant growth promotion. Table 5.2 represents QS
signals and its regulated functions.
Table 5.2 Activities of soilborne bacterial functions regulated by QS signals
98
Representative genus or
Taxonomic class species QS signals produced Known regulated functions References
Actinobacteria Streptomyces sp. Gamma-butyrolactones Antibiotic compound synthesis, Chater (1993) and Shaaban
differentiation et al. (2016)
Alpha- Agrobacterium tumefaciens OOHL Ti plasmid transfer, virulence White and Winans (2007)
proteobacteria Bradyrhizobium sp. AHLs Not determined Pongsilp et al. (2005)
Bradyoxetin Expression of nod genes Loh et al. (2002)
pCHL Not determined Schaefer et al. (2008)
Rhizobium leguminosarum OHtDeHL, HHL, HHL, Growth inhibition, induction of rhi genes, González and Marketon (2003)
pv. viciae OHL, OHOHL, OOHL plasmid transfer, nodulation
Sinorhizobium meliloti OHL, OHOHL, OOHL, EPSII synthesis, motility, nodulation González and Marketon (2003)
DDHL, OtdHL, HDHL, kinetics, plasmid transfer
OhDHL
Mesorhizobium loti OHHL, OHL, DHL, dDHL Nodulation Yang et al. (2009)
Rhodopseudomonas pCHL Chemotaxis Schaefer et al. (2008)
palustris
Beta- Burkholderia glumae OHL Toxoflavin synthesis Ferluga et al. (2008)
proteobacteria Burkholderia kururiensis OHHL, OHL, DHL, Not determined Ferluga et al. (2008)
dDHL
Ralstonia solanacearum OHL Not determined Ferluga et al. (2008)
Firmicutes Staphylococcus aureus AIPs (thiolactone Virulence, antimicrobial peptides synthesis, Sturme et al. (2007) and Lyon
peptides) genetic competence and Novick (2004)
Gamma- Erwinia amylovora OHHL Virulence, EPS synthesis, hydrogen Ferluga et al. (2008)
proteobacteria peroxide tolerance
Pectobacterium OHHL Varbapenem production, exoenzymes, hrp Braeken et al. (2008)
carotovorum secretion system
Dickeya dadantii HHL, OHHL, DHL Not determined Ferluga et al. (2008)
Pantoea stewartii OHHL, OOHL EPS synthesis, biofilm, adhesion, xylem Braeken et al. (2008)
dissemination, pathogenicity
R. M.P.
Several studies revealed that QS is playing a vital role in inducing protection against
abiotic factors, environmental challenges (Popat et al. 2015), and production of bio-
surfactant along with plant growth promotion. Biosurfactants produced by the rhi-
zosphere and plant-associated microbes are associated with rhizosphere activity,
plant microbe interaction, and plant pathogen elimination, increasing the bioavail-
ability of nutrient for beneficial plant-associated microbes. They are proven effec-
tive even in improving the agricultural soil quality through soil remediation.
Screening of biosurfactant-producing microorganisms from soil biosphere needs to
be accelerated, and the application of advanced methodologies such as functional
metagenomics will aid in obtaining unculturable biosurfactant-producing microbes
(Sachdev and Cameotra 2013). These studies add more insights into the understand-
ing of the QS mechanisms and their role in soil fertility enrichment.
Agriculture is one of the most significant factors contributing to the economic
growth of India, and sustainable agriculture economic development is the need of
the hour. Hence, a major focus in the coming decades would be on safe and eco-
friendly methods by exploiting the beneficial microorganisms in sustainable crop
production. Even genetic engineering of rhizosphere microflora and transgenic
plants which can alter quorum sensing in microbes will have several benefits in
agriculture including yield enhancement and effective plant pathogen management.
Besides the growing information on the production of growth regulators and com-
petitiveness of the microbes in the rhizosphere, microsymbionts, and other factors,
their effect upon plant growth will become more evident.
Of the ongoing emergence of antibiotic-resistant pathogens, there is a current
need for development of alternative management strategies. An antivirulence
approach by which quorum sensing is impeded is caught on as a viable means to
manipulate bacterial processes, especially pathogenic traits that are harmful to
human and animal health and agricultural productivity (LaSarre and Federle 2013).
The identification and development of chemical compounds and enzymes that facil-
itate quorum sensing inhibition by targeting signaling molecules, signal biogenesis,
or signal detection are gaining momentum.
Nowadays, screening for quorum quenching compounds is another interesting
area which can be carried out in high-throughput screenings since several assays
targeting the formation of autoinducers or their receptors are available (Abraham
2016; Reuter et al. 2016). However, the entire mechanism for quorum sensing is
known only for a small number of microorganisms, and still very little is known
about interspecies cross talks.
References
Abraham W-R (2016) Going beyond the control of quorum-sensing to combat biofilm infections.
Antibiotics 5:3–19
Ahmad M, Nadeem SM, Naveed M, Zahir ZA (2016) Potassium-solubilizing bacteria and their
application in agriculture. In: Meena VS, Maurya BR, Verma JP, Meena RS (eds) Potassium
5 Quorum Sensing in Plant Microbe Interaction 101
Chauhan PS, Nautiyal CS (2010) The pur B gene controls rhizosphere colonization by Pantoea
agglomerans. Lett Appl Microbiol 50:205–210
Chen X, Schauder S, Potier N, Van Dorssealaer A, Pelczer I, Bassler BL, Hughson FM (2002)
Structural identification of a bacterial quorum-sensing signal containing boron. Nature
415:545–549
Chernin L, Ismailov Z, Haran S, Chet I (1995) Chitinolytic Enterobacter agglomerans antagonistic
to fungal plant pathogens. Appl Environ Microbiol 61:1720–1726
Chin-A-Woeng TFC, Bloemberg GV, Mulders IHM, Dekkers LC, Lugtenberg BJJ (2000) Root
colonization by phenazine-1-carboxamide-producing bacterium Pseudomonas chlororaphis
PCL 1391 is essential for biocontrol of tomato foot and root rot. Mol Plant-Microbe Interact
13:1340–1345
Courchamp F, Clutton-Brock T, Grenfell B (1999) Inverse density dependence and the Allee effect.
Trends Ecol Evol 14:405–410
Crespi BJ (2001) The evolution of social behaviour in microorganisms. Trends Ecol Evol
16:178–183
Cui S, Meng J, Bhagwat A (2001) Availability of glutamate and arginine during acid challenge
determines cell density-dependent survival phenotype of Escherichia coli strains. Appl Environ
Microbiol 67:4914–4918
Darch SE, Stuart A, West SA, Winzer K, Diggle SP (2012) Density-dependent fitness benefits in
quorum-sensing bacterial populations. PNAS 109:8259–8263
Das I, Pradhan M (2016) Potassium-solubilizing microorganisms and their role in enhancing
soil fertility and health. In: Meena VS, Maurya BR, Verma JP, Meena RS (eds) Potassium
solubilizing microorganisms for sustainable agriculture. Springer, India, pp 281–291.
doi:10.1007/978-81-322-2776-2_20
Dastager SG, Deepa CK, Pandey A (2010) Isolation and characterization of novel plant growth
promoting Micrococcus sp. NII-0909 and its interaction with cowpea. Plant Physiol Biochem
48:987–992
DeAngelis KM, Firestone MK, Lindow SE (2007) A sensitive whole-cell biosensor suitable for
detecting a variety of n-acyl homoserine lactones in intact rhizosphere microbial communities.
Appl Environ Microbiol 73:3724–3727
DeAngelis KM, Lindow SE, Firestone MK (2008) Bacterial quorum sensing and nitrogen cycling
in rhizosphere soil. FEMS Microbiol Ecol 66:197–207
Diggle SP, Matthijs S, Wright VJ, Fletcher MP, Chhabra SR, Lamont IL, Kong X, Hider RC,
Cornelis P, Cámara M, Williams P (2007) The Pseudomonas aeruginosa 4-quinolone signal
molecules HHQ and PQS play multifunctional roles in quorum sensing and iron entrapment.
Chem Biol 14:87–96
Dominguez-Nunez JA, Benito B, Berrocal-Lobo M, Albanesi A (2016) Mycorrhizal fungi: role
in the solubilization of potassium. In: Meena VS, Maurya BR, Verma JP, Meena RS (eds)
Potassium solubilizing microorganisms for sustainable agriculture. Springer, India, pp 77–98.
doi:10.1007/978-81-322-2776-2_6
Dong YM, Iniguez AL, Triplett EW (2004) Nitrogen fixation in wheat provided by Klebsiella
pneumonia 342. Mol Plant-Microbe Interact 17:1078–1085
Dotaniya ML, Meena VD, Basak BB, Meena RS (2016) Potassium uptake by crops as well
as microorganisms. In: Meena VS, Maurya BR, Verma JP, Meena RS (eds) Potassium
solubilizing microorganisms for sustainable agriculture. Springer, India, pp 267–280.
doi:10.1007/978-81-322-2776-2_19
Dubern JF, Diggle SP (2008) Quorum sensing by 2-alkyl-4-quinolones in Pseudomonas aerugi-
nosa and other bacterial species. Mol BioSyst 4:882–888
Dunny GM, Leonard BA (1997) Cell-cell communication in gram-positive bacteria. Annu Rev
Microbiol 51:527–564
Dusane DH, Zinjarde SS, Venugopalan VP, RJ ML, Wever MW, Rahman PK (2010) Quorum
sensing; implication on rhamnolipid biosurfactant production. Biotechnol Genet Eng Rev
27:159–184
5 Quorum Sensing in Plant Microbe Interaction 103
Elasri M, Delorme S, Lemanceau P, Stewart G, Laue B, Glickmann E, Oger PM, Dessaux Y (2001)
Acyl-homoserine lactone production is more common among plant-associated Pseudomonas
spp. than among soil borne Pseudomonas spp. Appl Environ Microbiol 67:1198–1209
Ellis CN, Cooper VS (2010) Experimental adaptation of Burkholderia cenocepacia to onion
medium reduces host range. Appl Environ Microbiol 76:2387–2396
Fan B, Chen XH, Budiharjo A, Bleiss W, Vater J, Borriss R (2011) Efficient colonization of plant
roots by the plant growth promoting bacterium Bacillus amyloliquifaciens FZB42, engineered
to express green fluorescent protein. J Biotechnol 151:303–311
Federle MJ, Bassler BL (2003) Interspecies communication in bacteria. J Clin Invest 112:1291–1299
Ferluga S, Staindler L, Ventiru V (2008) N-acyl homoserine lactone quorum sensing in gram nega-
tive rhizobacteria. In: Karlovsky P (ed) Secondary metabolites in soil ecology, Soil biology, vol
14. Springer, Berlin, pp 69–92
Fleuchot B, Gitton C, Guillot A, Vidic J, Nicolas P, Besset C, Fontaine L, Hols P, Leblond-Bourget
N, Monnet V, Gardan R (2011) Rgg proteins associated with internalized small hydrophobic
peptides: a new quorum-sensing mechanism in Streptococci. Mol Microbiol 80:1102–1119
Fontaine L, Boutry C, de Frahan MH, Delplace B, Fremaux C, Horvath P, Boyaval P, Hols PA
(2010) Novel pheromone quorum-sensing system controls the development of natural compe-
tence in Streptococcus thermophilus and Streptococcus salivarius. J Bacteriol 192:1444–1454
Foster J (1995) Low pH adaptation and the acid tolerance response of Salmonella typhimurium.
Crit Rev Microbiol 21:215–237
Foster J, Hall H (1991) Inducible pH homeostasis and the acid tolerance response of Salmonella
typhimurium. J Bacteriol 173:5129–5135
Fray RG (2002) Altering plant-microbe interaction through artificially manipulating bacterial quo-
rum sensing. Ann Bot 89:245–253
Fujishige NA, Kapadia NN, De Hoff PL, Hirsch AM (2006) Investigations of Rhizobium biofilm
formation. FEMS Microbiol Ecol 56:195–206
Fuqua C, Greenberg EP (2002) Listening in on bacteria: acyl-homoserine lactone signalling. Nat
Rev Mol Cell Biol 3:685–695
González JE, Marketon MM (2003) Quorum sensing in nitrogen-fixing rhizobia. Microbiol Mol
Biol Rev 67:574–592
Guerra-Santos LH, Käppeli O, Fiechter A (1986) Dependence of Pseudomonas aeruginosa contin-
uous culture biosurfactant production on nutritional and environmental factors. Appl Microbiol
Biotechnol 24:443–448
Haggag WM, Timmusk S (2008) Colonization of peanut roots by biofilm forming Paenibacillus
polymyxa initiated biocontrol against crown rot disease. J Appl Microbiol 104(4):961–969
Hartmann A, Rothballer M, Hense BA, Schroder P (2014) Bacterial quorum sensing compounds
are important modulators of microbe-plant interactions. Front Plant Sci 5:1–4
Hawkes CV, DeAngelis KM, Firestone MK (2007) Root interactions with soil microbial commu-
nities and processes. In: Cardon Z, Whitbeck J (eds) The rhizosphere: an ecological perspec-
tive. Academic, New York, pp 1–30
He YW, Zhang LH (2008) Quorum sensing and virulence regulation in Xanthomonas campestris.
FEMS Microbiol Rev 32:842–857
Holden MT, Ram Chhabra S, de Nys R, Stead P, Bainton NJ, Hill PJ, Manefield M, Kumar N,
Labatte M, England D, Rice S, Givskov M, Salmond GP, Stewart GS, Bycroft BW, Kjelleberg
S, Williams P (1999) Quorum-sensing cross talk: isolation and chemical characterization
of cyclic dipeptides from Pseudomonas aeruginosa and other gram-negative bacteria. Mol
Microbiol 33:1254–1266
Huang JH, Elzinga EJ, Brechbuehl Y, Viegelin A, Kretzschmr R (2011) Impacts of Shewanella
putrefaciens strain CN-32 cells and extracellular polymeric substances on the sorption of
As(V) and As(III) on Fe(III)-hydroxides. Environ Sci Technol 45:2804–2810
Hughes DT, Sperandio V (2008) Inter-kingdom signalling: communication between bacteria and
their hosts. Nat Rev Microbiol 6:111–120
Ingham CJ, Jacob EB (2008) Swarming and complex pattern formation in Paenibacillus vortex
studied by imaging and tracking cells. BMC Microbiol 8:36
104 R. M.P.
Ingham PW, Nakano Y, Seger C (2011) Mechanisms and functions of Hedgehog signalling across
the metazoa. Nat Rev Genet 12:393–406
Jaiswal DK, Verma JP, Prakash S, Meena VS, Meena RS (2016) Potassium as an important plant
nutrient in sustainable agriculture: a state of the art. In: Meena VS, Maurya BR, Verma JP,
Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture. Springer,
India, pp 21–29. doi:10.1007/978-81-322-2776-2_2
Jakovleva J, Teppo A, Velts A, Sauma S, Moor H, Kivisar M, Teras R (2012) Fis regulates the
competitiveness of Pseudomonas putida on barley roots by inducing biofilm formation.
Microbiology 158(3):708–720
Janczarek M, Skorupska A (2011) Modulation of rosR expression and exopolysaccharide pro-
duction in Rhizobium leguminosarum pv. trifolii by clover root exudates. Int J Mol Sci
12:4132–4155
Jat LK, Singh YV, Meena SK, Meena SK, Parihar M, Jatav HS, Meena RK, Meena VS (2015)
Does integrated nutrient management enhance agricultural productivity? J Pure Appl Microbiol
9(2):1211–1221
Jha Y, Subramanian RB (2016) Regulation of plant physiology and antioxidant enzymes for alle-
viating salinity stress by potassium-mobilizing bacteria. In: Meena VS, Maurya BR, Verma JP,
Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture. Springer,
India, pp 149–162. doi:10.1007/978-81-322-2776-2_11
Ji G, Beavis RC, Novick RP (1995) Cell density control of staphylococcal virulence mediated by
an octapeptide pheromone. Proc Natl Acad Sci U S A 92:12055–12059
Ji X, Lu G, Gai Y, GAo H, Lu B, Kong B, Mu Z (2010) Colonization of Morus alba L. by the
plant-growth-promoting and antagonist bacterium Burkholderia cepacia strain Lu10-1. BMC
Microbiol 10:243–254
Kawabuchi M, Hara Y, Nihira T, Yamada Y (1997) Production of butyrolactone autoregulators by
Streptomyces coelicolor A3(2). FEMS Microbiol Lett 157:81–85
Kawaguchi T, Azuma M, Horinouchi S, Beppu T (1988) Effect of B-factor and its analogous on
rifamycin biosynthesis in Nocardia sp. J Antibiot 41:360–365
Kearns DB, Losick R (2003) Swarming motility in undomesticated Bacillus subtilis. Mol
Microbiol 49:581–590
Kennedy A (2005) Rhizosphere. In: Sylvia D, Fuhrmann J, Hartel P, Zuberer D (eds) Principles
and applications of soil microbiology, 2nd edn. Pearson Prentice Hall, Upper Saddle River,
pp 242–262
Kent AD, Triplett EW (2002) Microbial communities and their interactions in soil and rhizosphere
ecosystems. Annu Rev Microbiol 56:211–236
Khan N, Mishra A, Chauhan PS, Sharma YK, Nautiyal CS (2012) Paenibacillus lentimorbus
enhances growth of chickpea (Cicer arietinum L.) in chromium-amended soil. Antonie Van
Leeuwenhoek 101:453–459
Kim C, Kecskes ML, Deaker RJ, Gilchrist K, New PB, Kennedy IR, Kim S, Sa T (2005) Wheat
root colonization and nitrogenase activity by Azospirillum isolates from crop plants in Korea.
Can J Microbiol 51:948–956
Köhler T, Curty LK, Barja F, van Delden C, Pechère J-C (2000) Swarming of Pseudomonas
aeruginosa is dependent on cell-to-cell signaling and requires flagella and pili. J Bacteriol
182(21):5990–5996
Kumar R, Bhatia R, Kukreja K, Behl RK, Dudeja SS, Narula N (2007) Establishment of Azotobacter
on plant roots: chemotactic response, development and analysis of root exudates of cotton
(Gossypium hirsutum L.) and wheat (Triticum aestivum L.) J Basic Microbiol 47:436–439
Kumar A, Bahadur I, Maurya BR, Raghuwanshi R, Meena VS, Singh DK, Dixit J (2015) Does
a plant growth-promoting rhizobacteria enhance agricultural sustainability? J Pure Appl
Microbiol 9:715–724
Kumar A, Meena R, Meena VS, Bisht JK, Pattanayak A (2016a) Towards the stress management
and environmental sustainability. J Clean Prod 137:821–822
Kumar A, Patel JS, Bahadur I, Meena VS (2016b) The molecular mechanisms of KSMs for
enhancement of crop production under organic farming. In: Meena VS, Maurya BR, Verma JP,
5 Quorum Sensing in Plant Microbe Interaction 105
Meena VS, Maurya BR, Bahadur I (2014a) Potassium solubilization by bacterial strain in waste
mica. Bang J Bot 43:235–237
Meena VS, Maurya BR, Verma JP (2014b) Does a rhizospheric microorganism enhance K+ avail-
ability in agricultural soils? Microbiol Res 169:337–347
Meena RS, Meena VS, Meena SK, Verma JP (2015a) The needs of healthy soils for a healthy
world. J Clean Prod 102:560–561
Meena RS, Meena VS, Meena SK, Verma JP (2015b) Towards the plant stress mitigate the agricul-
tural productivity: a book review. J Clean Prod 102:552–553
Meena VS, Maurya BR, Meena RS (2015c) Residual impact of wellgrow formulation and NPK on
growth and yield of wheat (Triticum aestivum L.) Bangladesh J Bot 44(1):143–146
Meena VS, Maurya BR, Verma JP, Aeron A, Kumar A, Kim K, Bajpai VK (2015d) Potassium
solubilizing rhizobacteria (KSR): isolation, identification, and K-release dynamics from waste
mica. Ecol Eng 81:340–347
Meena VS, Meena SK, Verma JP, Meena RS, Ghosh BN (2015e) The needs of nutrient use efficiency
for sustainable agriculture. J Clean Prod 102:562–563. doi:10.1016/j.jclepro.2015.04.044
Meena VS, Verma JP, Meena SK (2015f) Towards the current scenario of nutrient use efficiency in
crop species. J Clean Prod 102:556–557. doi:10.1016/j.jclepro.2015.04.030
Meena RK, Singh RK, Singh NP, Meena SK, Meena VS (2016a) Isolation of low temperature sur-
viving plant growth-promoting rhizobacteria (PGPR) from pea (Pisum sativum L.) and docu-
mentation of their plant growth promoting traits. Biocatalysis Agric Biotechnol 4:806–811
Meena RS, Bohra JS, Singh SP, Meena VS, Verma JP, Verma SK, Sihag SK (2016b) Towards the
prime response of manure to enhance nutrient use efficiency and soil sustainability a current
need: a book review. J Clean Prod 112(1):1258–1260
Meena SK, Rakshit A, Meena VS (2016c) Effect of seed bio-priming and N doses under varied
soil type on nitrogen use efficiency (NUE) of wheat (Triticum aestivum L.) under greenhouse
conditions. Biocatal Agric Biotechnol 6:68–75
Meena VS, Bahadur I, Maurya BR, Kumar A, Meena RK, Meena SK, Verma JP (2016d) Potassium-
solubilizing microorganism in evergreen agriculture: an overview. In: Meena VS, Maurya BR,
Verma JP, Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture.
Springer, India, pp 1–20. doi:10.1007/978-81-322-2776-2_1
Meena VS, Meena SK, Bisht JK, Pattanayak A (2016e) Conservation agricultural practices in
sustainable food production. J Clean Prod 137:690–691
Mendelson NH, Salhi B (1996) Patterns of reporter gene expression in the phase diagram of
Bacillus subtilis colony forms. J Bacteriol 178:1980–1989
Miller MB, Bassler BL (2001) Quorum sensing in bacteria. Annu Rev Microbiol 55:165–199
Munch C, Neu T, Kuschk P, Roske I (2016) The root surface as the definitive detail for microbial
transformation processes in constructed wetlands – a biofilm characteristic. Water Sci Technol
56(3):271–276
Nath D, Maurya BR, Meena VS (2017) Documentation of five potassium- and phosphorus-
solubilizing bacteria for their K and P-solubilization ability from various minerals. Biocatal
Agric Biotechnol 10:174–181
Neumann G (2007) Root exudates and nutrient cycling. In: Marschner P, Rengel Z (eds) Nutrient
cycling in terrestrial ecosystems. Springer, Berlin, pp 123–157
Newton JA, Fray RG (2004) Integration of environmental and host derived signals with quorum
sensing during plant–microbe interactions. Cell Microbiol 6:213–224
Nihorimbere V, Marc Ongena M, Smargiassi M, Thonart P (2011) Beneficial effect of the rhi-
zosphere microbial community for plant growth and health. Biotechnol Agron Soc Environ
15:327–337
Nihorimbere V, Cawoy H, Seyer A, Brunelle A, Thonart P, Ongena M (2012) Impact of rhizo-
sphere factors on cyclic lipopeptide signature from the plant beneficial strain Bacillus amylo-
liquefaciens S499. FEMS Microbiol Ecol 79:176–191
Novick RP (2003) Autoinduction and signal transduction in the regulation of staphylococcal viru-
lence. Mol Microbiol 48:1429–1449
5 Quorum Sensing in Plant Microbe Interaction 107
Ryan RP, Fouhy Y, Garcia BF, Watt SA, Niehaus K, Yang L, Tolker-Nielsen T, Dow JM (2008)
Interspecies signaling via the Stenotrophomonas maltophilia diffusible signal factor influences
biofilm formation and polymyxin tolerance in Pseudomonas aeruginosa. Mol Microbiol 68:75–86
Sachdev DP, Cameotra SS (2013) Biosurfactants in agriculture. Appl Microbiol Botechnol
97(3):1005–1016
Saha M, Maurya BR, Bahadur I, Kumar A, Meena VS (2016a) Can potassium-solubilising bac-
teria mitigate the potassium problems in India? In: Meena VS, Maurya BR, Verma JP, Meena
RS (eds) Potassium solubilizing microorganisms for sustainable agriculture. Springer, India,
pp 127–136. doi:10.1007/978-81-322-2776-2_9
Saha M, Maurya BR, Meena VS, Bahadur I, Kumar A (2016b) Identification and characterization
of potassium solubilizing bacteria (KSB) from Indo-Gangetic Plains of India. Biocatal Agric
Biotechnol 7:202–209
Sandhya V, Ali SKZ, Grover M, Reddy G, Venkateswarlu B (2009) Alleviation of drought stress
effects in sunflower seedlings by the exopolysaccharides producing Pseudomonas putida strain
GAP-P45. Biol Fertil Soils 46(1):17–26
Sarkar D, Meena VS, Haldar A, Rakshit R (2017) Site-specific nutrient management (SSNM):
a unique approach towards maintaining soil health. In the Adaptive soil management: from
theory to practices, p 69–88. doi:10.1007/978-981-10-3638-5_3
Schaefer AL, Greenberg EP, Oliver CM, Oda Y, Huang JJ, Bittan-Banin G, Peres CM, Schmidt
S, Juhaszova K, Sufrin JR, Harwood CS (2008) A new class of homoserine lactone quorum-
sensing signals. Nature 454:595–599
Schauder S, Bassler BL (2001) The languages of bacteria. Genes Dev 15:1468–1480
Schue M, Fekete A, Ortet P, Brutesco C, Heulin T, Schmitt-Kopplin P, Achiuak W, Santelia C
(2011) Modulation of metabolism and switching to biofilm prevail over exopolysaccharide
production in the response of Rhizobium alamii to cadmium. PLoS One 611:e26771
Shaaban MI, Bar FMA, El-Mahdy AM, Shokralla S (2016) Quorum sensing inhibiting activity of
Streptomyces coelicoflavus isolated from soil. Front Microbiol 7:659–671
Shankar M, Ponraj P, Ilakkiam D, Gunasekaran P (2011) Root colonization of rice growth promot-
ing strain of Enterobacter cloacae. J Basic Microbiol 51(5):523–530
Sharma A, Sahgal M, Johri BN (2003) Microbial communication in the rhizosphere: operation of
quorum sensing. Curr Sci 85:1164–1172
Sharma A, Shankhdhar D, Shankhdhar SC (2016) Potassium-solubilizing microorganisms:
mechanism and their role in potassium solubilization and uptake. In: Meena VS, Maurya BR,
Verma JP, Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture.
Springer, India, pp 203–219. doi:10.1007/978-81-322-2776-2_15
Sheludko AV, Shirokov AA, Sokolova MK, Sokolov OI, Petrova LP, Matora LY, Katsy EI
(2010) Wheat root colonization by Azospirillum brasilense strains with different motility.
Microbiology 9(5):6887–6695
Shen X, Chen M, Hu H, Wang W, Peng H, Xu P, Zhang X (2012) Genome sequence of Pseudomonas
chlororaphis GP72, a root-colonizing biocontrol strain. J Bacteriol 194(5):1269–1270
Shrivastava M, Srivastava PC, D’Souza SF (2016) KSM soil diversity and mineral solubilization,
in relation to crop production and molecular mechanism. In: Meena VS, Maurya BR, Verma JP,
Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture. Springer,
India, pp 221–234. doi:10.1007/978-81-322-2776-2_16
Sigler WV, Nakatsu CH, Reicher ZJ, Turco RR (2001) Fate of the biological control agent Pseudomonas
aureofaciens TX-1 after application to turfgrass. Appl Environ Microbiol 67:3542–3548
Silby MW, Levy SB (2004) Use of in vivo expression technology to identify genes important
in growth and survival of Pseudomonas fluorescens Pf0-1 in soil: discovery of expressed
sequences with novel genetic organization. J Bacteriol 186:7411–7419
Simao-Beaunoir A-M, Roy S, Beaulieu C (2009) Microbial traits associated with actinobacteria
interacting with plants. In: Bouarab K, Brisson N, Daayf F (eds) Molecular plant–microbe
interactions. CAB International, USA, p 299
Sindhu SS, Parmar P, Phour M, Sehrawat A (2016) Potassium-solubilizing microorganisms
(KSMs) and its effect on plant growth improvement. In: Meena VS, Maurya BR, Verma JP,
5 Quorum Sensing in Plant Microbe Interaction 109
Verma JP, Jaiswa DK, Meena VS, Meena RS (2015a) Current need of organic farming for enhanc-
ing sustainable agriculture. J Clean Prod 102:545–547
Verma JP, Jaiswal DK, Meena VS, Kumar A, Meena RS (2015b) Issues and challenges about sus-
tainable agriculture production for management of natural resources to sustain soil fertility and
health. J Clean Prod 107:793–794
Verma R, Maurya BR, Meena VS, Dotaniya ML, Deewan P (2017a) Microbial dynamics as influ-
enced by bio-organics and mineral fertilizer in alluvium soil of Varanasi, India. Int J Curr
Microbiol App Sci 6(2):1516–1524
Verma R, Maurya BR, Meena VS, Dotaniya ML, Deewan P, Jajoria M (2017b) Enhancing pro-
duction potential of cabbage and improves soil fertility status of Indo-Gangetic Plain through
application of bio-organics and mineral fertilizer. Int J Curr Microbiol App Sci 6(3):301–309
Walker TS, Bais HP, Grotewold E, Vivanco JM (2003) Root exudation and rhizosphere biology.
Plant Physiol 132:44–51
Waters CM, Bassler BL (2005) Quorum sensing: cell-to-cell communication in bacteria. Annu Rev
Cell Dev Biol 21:319–346
Waters CM, Bassler BL (2006) The Vibrio harveyi quorum-sensing system uses shared regulatory
components to discriminate between multiple autoinducers. Genes Dev 20:2754–2767
Wei HL, Zhang LQ (2006) Quorum-sensing system influences root colonization and biological
control ability in Pseudomonas fluorescens 2P24. Antonie Van Leeuwenhoek 89:267–280
White CE, Winans SC (2007) Cell-cell communication in the plant pathogen Agrobacterium tume-
faciens. Philos Trans R Soc Lond Ser B Biol Sci 362:1135–1148
Williams A, Wilkinson A, Krehenbrink M, Russo DM, Zorreguieta A, Downie JA (2008)
Glucomannan-mediated attachment of Rhizobium leguminosarum to pea root hairs is required
for competitive nodule infection. J Bacteriol 190:4706–4715
Williamson LL, Borlee BR, Schloss PD, Guan C, Allen HK, Handelsman J (2005) Intracellular
screen to identify metagenomic clones that induce or inhibit a quorum-sensing biosensor. Appl
Environ Microbiol 71:6335–6344
Yadav BK, Sidhu AS (2016) Dynamics of potassium and their bioavailability for plant nutrition.
In: Meena VS, Maurya BR, Verma JP, Meena RS (eds) Potassium solubilizing microorganisms
for sustainable agriculture. Springer, India, pp 187–201. doi:10.1007/978-81-322-2776-2_14
Yamada Y, Nihira T (1998) Microbial hormones and microbial chemical ecology. In: Barton DHR,
Nakanishi K (eds) Comprehensive natural products chemistry, vol 8. Elsevier, Oxford, pp 377–413
Yamada Y, Sugamura K, Kondo K, Yanagimoto M, Okada H (1987) The structure of inducing fac-
tors for virginiamycin production in Streptomyces virginias. J Antibiot 40:496–504
Yamaga F, Washio A, Morikawa M (2010) Sustainable biodegradation of phenol by Acinetobacter
calcoaceticus P23 isolated from the rhizosphere of duckweed Lemna aoukkiusa. Environ Sci
Technol 44:470–6474
Yang J, Kharbanda PD, Mirza M (2004) Evaluation of Paenibacillus polymyxa pkb1for biocontrol
of Pythium disease of cucumber in a hydroponic system. Acta Horticult 635:59–66
Yang M, Sun K, Zhou L, Yang R, Zhong Z, Zhu J (2009) Functional analysis of three AHL auto-
inducer synthase genes in Mesorhizobium loti reveals the important role of quorum sensing in
symbiotic nodulation. Can J Microbiol 55:210–214
Yasin M, Munir I, Faisal M (2016) Can Bacillus spp. enhance K+ uptake in crop species. In:
Meena VS, Maurya BR, Verma JP, Meena RS (eds) Potassium solubilizing microorganisms
for sustainable agriculture. Springer, India, pp 163–170. doi:10.1007/978-81-322-2776-2_12
Zahavi A, Ralt D (1984) Social adaptation in myxobacteria. In: Rosenberg E (ed) Myxobacteria:
development and cell interactions. Springer-Verlag, New York, pp 215–220
Zahedi H (2016) Growth-promoting effect of potassium-solubilizing microorganisms on
some crop species. In: Meena VS, Maurya BR, Verma JP, Meena RS (eds) Potassium
solubilizing microorganisms for sustainable agriculture. Springer, India, pp 31–42.
doi:10.1007/978-81-322-2776-2_3
Horizontal Gene Transfer in Soil
and the Rhizosphere: Impact 6
on Ecological Fitness of Bacteria
Meenu Maheshwari, Hussein H. Abulreesh,
Mohammad Shavez Khan, Iqbal Ahmad, and John Pichtel
Abstract
The ecological fitness of soil- and root-associated bacterial communities is a key
element for soil fertility and plant health as well as plant stress tolerance. Genetic
variability in bacterial populations is maintained through mutation and gene
acquisition. Horizontal gene transfer (HGT) is accomplished by conjugation,
transformation, and transduction both in vitro and under natural conditions.
Mobile genetic elements (MGEs) play a significant role in gene dissemination in
bacterial communities and increase their adaptability, survival, and ability to
colonize different environmental niches. In this context, bacterial conjugative
plasmids encoding resistance genes, degradative genes, and tolerance to stress
conditions are of much significance. The biofilm mode of bacterial growth fur-
ther enhances gene exchange and increase the fitness and competitiveness of
bacteria. Microcosm studies reveal a number of factors influencing the HGT pro-
cess in soil. Considering the importance of HGT, a better understanding of
genetic processes in the rhizosphere will further help in effective exploitation of
naturally engineered bacteria for sustainable agriculture.
Keywords
Bacteria • Horizontal gene transfer • Conjugative plasmid • Microcosm • Biofilm
• Rhizosphere • Ecological fitness
6.1 Introduction
Horizontal gene transfer (HGT) refers to the transfer of genetic material between
prokaryotes. The process occurs in one of three ways: (1) transformation (the uptake
of free DNA by competent bacteria from the surrounding environment), (2) trans-
duction (gene transfer mediated by bacteriophages), and (3) conjugation (gene
transfer by means of plasmids or integrative conjugative elements). Conjugation is
the process by which a DNA molecule (i.e., plasmid or conjugative transposon) is
transferred from a donor cell to a physically attached recipient cell via some conju-
gation apparatus (Zechner et al. 2000). Although most conjugative plasmids shared
common mechanistic principles, e.g., synthesis of conjugative pili, there is a remark-
able diversity of conjugative systems in Gram-negative and Gram-positive bacteria,
depending on the shape and other characteristics of the plasmid-encoded pili.
Nonconjugative plasmids are transferred to recipient cells by mobilizing/helper
plasmids (Heuer and Smalla 2007). Transfer of conjugative plasmids or transposons
has been demonstrated to occur in various ecological habitats, for example, the
plant surface, rhizosphere, surface water, and human and animal intestines (Aminov
2011; Madsen et al. 2012; Huddleston 2014; Juhas 2015; Pinto-Carbo et al. 2016;
Sun et al. 2016). In contrast to conjugation, transduction provides a means of DNA
acquisition in which nonviral DNA is transferred from an infected host bacterium to
a new host via infectious or noninfectious virus particles (Meena et al. 2013a;
Bahadur et al. 2014; Maurya et al. 2014; Jaiswal et al. 2016; Jha and Subramanian
2016; Kumar et al. 2016a, b).
Defective phage particles released from lysed host cells attach to new host cells
and deliver their DNA into the new host. The injected bacterial DNA is subse-
quently integrated into the recipient genome. Unlike conjugation, transduction does
not require cell-to-cell contact; however, most bacteriophages infect only a narrow
range of hosts (Wommack and Colwell 2000). Evidence for the importance of trans-
duction as an HGT process in the natural environment arises from studies on the
abundance of bacteriophages in different settings, primarily soil (Ashelford et al.
2003) and from bacterial genome sequences (Canchaya et al. 2003). The role of
bacteriophages in contributing various genes to bacterial genera of medical and
ecological significance is well documented (Weitz et al. 2013; Broszat and
Grohmann 2014; Dalmasso et al. 2014; Chen et al. 2015; Obeng et al. 2016).
Horizontal gene transfer plays an important role in formulating bacterial
genomes, promoting intra- and interspecies variability and distributing functional
genetic modules within communities. HGT provides a means for enhanced under-
standing of ecological adaptation and bacterial evolution in the biosphere. Extensive
gene exchange during bacterial evolution is made evident by the marked similarity
6 Horizontal Gene Transfer in Soil and the Rhizosphere: Impact on Ecological… 113
of genes among distantly related species, variation of gene content between closely
related strains, and incongruent phylogenetic trees. Horizontal gene transfer is a
major force in bacterial evolution; a well-documented example of HGT is the spread
of antibiotic resistance genes among pathogenic and nonpathogenic bacteria
(Barlow 2009; Boto 2010; Davies and Davies 2010; Abulreesh 2011, 2012; Baltrus
2013; Wellington et al. 2013). The driving force for the acquisition of foreign genes
via HGT is believed to be the need for bacteria to overcome environmental stresses
for survival and to compete successfully in their ecological niches (Hacker and
Kaper 2000).
Mobile genetic elements (MGEs) such as plasmids, bacteriophages, integrative
conjugative elements, transposons, insertion sequences (IS) elements, integrons,
gene cassettes, and genomic islands are the key vehicles among HGT mechanisms.
In many species, a high proportion of horizontally transferred genes can be attrib-
uted to plasmid-, phage-, or transposon-related sequences, as remnants of these
mobile elements have been located adjacent to genes identified as horizontally
transferred (Ochman et al. 2000; Brussow et al. 2004; Frost et al. 2005; Gyles and
Boerlin 2013). It has been suggested that MGEs add some metabolic burden to their
host, and adaptation occurs to minimize this impact (Dahlberg and Chao 2003;
Heuer et al. 2007). The prevalence of plasmids, however, indicates that they benefit
bacteria and compensate for any burden they might impose on the cell. Plasmids
persist because bacterial communities and their environments are continuously
changing; the variability carried by these genetic elements increases the speed at
which adapted strains arise, and the adapted strains retain the MGE so they can
propagate rapidly (Jat et al. 2015; Kumar et al. 2015; Ahmad et al. 2016; Meena
et al. 2015f, 2016a; Dominguez-Nunez et al. 2016; Dotaniya et al. 2016).
The MGEs that enhance an organism’s adaptability evolve and survive at the
expense of those that do not. Thus, MGEs confer an improved fitness to the bacterial
community and its ability to colonize different environmental niches. MGEs addi-
tionally increase the possibility of new strains arising with novel or increased selec-
tive advantages over neighboring communities. Metagenomic approaches have
revealed a large and untapped diversity of resident MGEs in soil- and plant-
associated bacteria. Approximately 18% of bacterial isolates from the phytosphere
of sugar beets was found to contain plasmids (Powell et al. 1993), and a large pro-
portion were able to mobilize non-self-transferable but mobilizable Inc-Q plasmids
(Kobayashi and Bailey 1994). The extensive presence of plasmid-related sequences
in soil DNA (directly extracted) indicates a significant abundance of plasmids in soil
(Heuer et al. 2009). The abundance of transferable plasmids in soil is believed to be
related to the presence of contaminants (e.g., antibiotics, xenobiotics). Several stud-
ies have indicated that genes encoding the enzymes involved in catabolism of envi-
ronmental pollutants are present on plasmids (Smets and Barkay 2005; Kopmann
et al. 2013).
Different methods involving various molecular techniques have been employed
to detect and study soil plasmid structure, organization and function (Heuer and
Smalla 2012). Moreover, metagenomics methods such as pyrosequencing of soil
DNA have proved to be highly sensitive and have led to the discovery of novel
114 M. Maheshwari et al.
plasmid sequences (Kristiansson et al. 2011). Genome sequencing data have
revealed the modular character of conjugative plasmids, where different modules
(e.g., a compactly arranged gene) are dedicated toward carrying out specific func-
tions (de la Cueva-Méndez and Pimentel 2007; Norman et al. 2009). The overall
genetic makeup of conjugative plasmids can be divided into four distinct categories
of modules: (1) modules affecting plasmid replication and copy control, (2) mod-
ules affecting plasmid stability, (3) modules affecting plasmid propagation, and (4)
modules affecting plasmid host adaptation, along with numerous other subcatego-
ries (Norman et al. 2009). The first three categories are considered “plasmid core”
or plasmid “backbone” genes. Sequence analysis of plasmid backbone genes is con-
sidered a consistent and reliable foundation for plasmid classification, as opposed to
traditionally used methods (Heuer and Smalla 2012). Apart from plasmid backbone
genes, a highly diverse, accessory, or flexible set of genes within soil plasmids
occurs as a fourth module which carries genes responsible for environmental adap-
tation. These accessory genes often differ greatly among plasmids with identical
plasmid backbones and impart the ability to adapt to varied environmental condi-
tions such as exposure to antibiotics, heavy metals, and xenobiotics (Dennis 2005;
Schluter et al. 2007; Heuer et al. 2009).
Degradative genes resident on MGEs have been isolated successfully from soil
treated with the herbicide 2, 4-D but not from untreated control soil. Similarly, self-
transferable plasmids which confer resistance toward antibiotics have been isolated
from animal manures used for soil fertilization (Heuer and Smalla 2007; van
Overbeek et al. 2002). Sequencing of plant-associated bacteria reveal that many
phytopathogenic and symbiotic bacteria carry plasmids (Zhao et al. 2005; Bardaji
et al. 2011), pathogenicity islands (Gardiner et al. 2014), or integrons (Gillings et al.
2005). These studies reveal the significance of horizontal gene transfer in bacterial
adaptation and evolution under changing environmental conditions (Parewa et al.
2014; Prakash and Verma 2016; Meena et al. 2015e, 2016b; Teotia et al. 2016;
Bahadur et al. 2016b; Das and Pradhan 2016).
In recent years, the importance of biofilm formation and its relationship to gene
transfer has received significant attention (Burmolle et al. 2014; Stalder and Top
2016). It is well recognized that horizontal gene transfer via plasmids occurs more
effectively on surfaces, e.g., in biofilms than among planktonic cells. Biofilms are
highly structured bacterial communities embedded in a self-produced matrix com-
posed of exopolysaccharides (EPSs), proteins, and DNA. These films adhere to bio-
logical and non-biological surfaces (Hall-Stoodley et al. 2004) and provide a
favorable environment for genetic elements to be transferred horizontally.
Rhizobacterial biofilms associated with plant roots support bacterial survival and
host plant colonization, reduce biotic and abiotic plant stress, and enhance agricul-
tural productivity (Lopez et al. 2010; Yadav and Sidhu 2016; Meena et al. 2016d;
Saha et al. 2016b; Verma et al. 2014, 2015b). All these advantages conferred by
biofilms are directly or indirectly associated with a high frequency of horizontal
gene transfer in the biofilm mode of growth. In this chapter, we focus on horizontal
gene transfer in soil- or plant-associated (rhizosphere) bacteria that contribute to
6 Horizontal Gene Transfer in Soil and the Rhizosphere: Impact on Ecological… 115
Fig. 6.1 Gene transfer in the rhizosphere: mechanisms, factors, and outcomes
On the other hand, considerable evidence indicates that HGT is an ongoing process
that plays a primary role in real-time ecological adaptations of prokaryotes (Smets
and Barkay 2005). MGEs play an essential role in the process by shaping the bacte-
rial genome, promoting intraspecies variability, and distributing functional genetic
modules among communities. Consequently, HGT of genetic modules that allows
adaptation to rapidly changing biotic interactions has frequently been observed
(Smets and Barkay 2005). The interactions include:
The most widely studied examples of genetic exchange through HGT in natural
environments is the dissemination of multiple antibiotic resistance via MGE, which
allows bacterial populations to adapt rapidly to strong selective pressures (Cordero
et al. 2012; Wellington et al. 2013; Ojala et al. 2014). MGEs involved in transmis-
sion of antibiotic resistance include combinatorial genetic evolution of MDR facili-
tated by transposons, IS elements, and integrons. Transposable elements like ISCRs
(insertion sequence common regions) mobilize DNA adjacent to their insertion site
via rolling circle replication (Toleman et al. 2006). ISCRs are closely associated
with antibiotic resistance genes on conjugative plasmids (Priyadharsini and
Muthukumar 2016; Kumar et al. 2017; Masood and Bano 2016; Meena et al. 2016e).
It is interesting to observe that under natural conditions, especially in wastewater
and soil, the presence of several toxic pollutants (e.g., metals and pesticides) may
increase selective pressure and co-selection of resistant strains. These pollutants
may further increase the ecological fitness and survivability of bacterial strains (De
Lipthay et al. 2008; Imfeld and Vuilleumier 2012). Various degradative plasmids
have been characterized from bacteria which degrade PCBs, chloroaniline, and
other recalcitrant molecules (Merlin et al. 1999; Boon et al. 2001; Springael and Top
2004). Many self-transferable plasmids participate directly in active gene transfer.
Russell et al. (2011) reviewed the evolutionary trends of enzymatic machinery
involved in the degradation of xenobiotics and concluded that horizontal gene trans-
fer among bacteria is one of the major determinants in the acquisition of new and
efficient enzymatic functions. New enzymatic pathways have emerged from a wide
variety of enzyme families.
Metagenomic mapping studies for two discretely located hexachlorocyclohex-
ane (HCH)-degrading strains of Sphingobium japonicum (Sphingobium japonicum
UT26 from Japan and Sphingobium indicum B90A from India) have shown that the
previous common ancestor was unable to degrade HCH isomers, but descendants
acquired degrading genes by transposon-mediated HGT (Sangwan et al. 2014).
Likewise, Pearce et al. (2015) showed that different HCH-degrading bacteria
6 Horizontal Gene Transfer in Soil and the Rhizosphere: Impact on Ecological… 117
yielded a distinct set of metabolites during degradation of HCH isomers. The analy-
sis confirmed the role of horizontal transfer mediated by an insertion sequence in
the acquisition of the pathway.
Among the conditions occurring within the phytosphere, the rhizosphere is opti-
mally suited for the growth and intensive interaction among microbial communities.
The availability of essential elements (C, N, P, K), moisture, and other critical con-
stituents (e.g., organic acids) in the vicinity of plant roots make the rhizosphere the
preferred site for gene transfer, such that it is referred to as a “hotspot” for genetic
evolution (van Elsas et al. 2003). The rhizosphere provides an ideal environment for
HGT processes and may support rapid adaptation of bacteria against environmental
changes as compared with bulk soil. Conjugative plasmid transfer frequency
between bacterial species is enhanced under high nutrient availability and water
movement (Kroer et al. 1998). Bacterial colonization and adherence to the root
surface in response to the presence of root exudates impart significant effects on
gene transfer.
Due to intimate contact among cells and higher metabolic activity in response to
the presence of an ample nutrient supply, elevated plasmid transfer in the rhizo-
sphere has been observed (Kroer et al. 1998; van Elsas et al. 2003). Musovic et al.
(2006) demonstrated the exceptionally broad host range of the pKJK10 plasmid in
the barley rhizosphere. They describe the potential of mobile genetic elements,
crossing large phylogenetic distances, i.e., gene swapping between Actinobacteria
and Proteobacteria, in bacterial evolution. Organic acids and other constituents of
root exudates stimulate the transformation in Acinetobacter sp. BD413 (pFG4) in
sterile soils. The organic acids, acetate, lactate, and alanine, resulted in the highest
transformation frequencies (Nielsen and van Elsas 2001). Molbak et al. (2007) stud-
ied conjugal transfer in the rhizospheres of pea and barley. The experiments showed
that a higher rate of root exudation and root growth rate in pea was responsible for
higher conjugal frequency. The distribution pattern of donors on pea roots was
shown to affect genetic transfer. Transfer of the plasmid RP4 derivative between
Pseudomonas fluorescens and Serratia sp. was studied in a sand microcosm and
found to be related to the availability of root exudates and bacterial metabolic activ-
ity (Kroer et al. 1998). Regardless, however, the direct relationship between the
metabolic activity of transforming bacteria and gene transfer is in need of further
investigation (Meena et al. 2013c, 2015a; Raghavendra et al. 2016; Zahedi 2016;
Singh et al. 2015; Bahadur et al. 2016a).
Genetic transfer of various contaminant-degrading genes has also been shown to
take place effectively in the rhizosphere and is concurrently responsible for enhanced
plant growth (Wang et al. 2007, 2014a, b; Jia et al. 2013). Using an in silico metage-
nomic prediction method, Cabezon et al. (2015) and Lopes et al. (2016) revealed a
significantly higher abundance of predicted genes associated with HGT in the rhi-
zosphere compared with bulk soil. The pilus assembly protein CpaE and type IV
118 M. Maheshwari et al.
Genetic variability among bacterial populations occurs via mutation and acquisition
of new genes through various genetic exchange mechanisms. Gene transfer in a
single species results in the propagation of specific traits. Interspecies gene transfer
may result in an entirely new genetic combination which may be of significance to
human health and the environment. The relationship between biofilms and HGT has
been investigated by numerous workers (Ghigo 2001; Tormo et al. 2005; Antonova
and Hammer 2011; Madsen et al. 2012; Cook and Dunny 2014). In general, HGT
rates are higher in biofilm communities compared with those in the planktonic state.
Biofilms are also implicated in the promotion of plasmid stability and may enhance
the host range of MGEs that are being transferred (Madsen et al. 2012; Broszat and
Grohmann 2014).
Several authors have reported enhanced HGT in biofilms via conjugation
(Sorensen et al. 2005; Maheshwari et al. 2016); however, certain spatial constraints
within biofilms may hinder the dispersal of plasmids (Krol et al. 2011; Merkey et al.
2011). Lili et al. (2007) indicated that plasmids which are maintained through high
transfer frequencies may only be able to persist in biofilms. Interestingly, other
mechanisms of gene transfer, for example, transformation, occur at higher rates in
biofilms. Gene transfer via both small DNA fragments as well as plasmid transfor-
mation has been documented (Maeda et al. 2006; Etchuuya et al. 2011). It has been
6 Horizontal Gene Transfer in Soil and the Rhizosphere: Impact on Ecological… 119
transfer in soil using microcosms with special focus on transfer of broad host range
plasmids and catabolic soil plasmids. The importance of gene transfer in soil and
other natural habitats has been recognized due to (a) interest in the possible spread
of genetically modified microorganisms, (b) spread and exchange of antibiotic
resistance, (c) the role of HGT in the evolution of new bacterial traits, and (d) the
possible role of introducing catabolic genes to enhance bioaugmentation strategies
to enhance bioremediation of soil contaminants.
Laboratory microcosms are used to assess gene transfer for a number of potential
benefits as mentioned above. Although microcosms are not exact replicas of the soil
system, they possess many of the same chemical and physical attributes. Microcosms
must be calibrated to ensure that they reflect natural conditions (Bolton et al. 1991).
Microcosms can be used to obtain vital information on (1) survival and dispersal of
bacterial strains, (2) ability to compete with indigenous microflora, (3) capacity to
exchange genetic information, and (4) stability of heterogeneous DNA in soil. Most
of the well-studied examples mainly comprise various conjugative plasmids and
their exchange by conjugation using suitable recipient strains. Some of the widely
used plasmids described by Hill and Tops (1998) are RP4, RP4p (RP4::pat),
pBLK1-2 (pRK2073::Tn5), RP4::Tn4371, pJP4, pEMT3k (pEMT3::mini Tn5),
pEMT1k (pEMT1::miniTn) R57.b, R388::Tn1721, pLF40, pFT30, pJB5JI::Tn5,
and pIJ673. Other plasmids studied include fluorescent marker-tagged plasmids
such as pB10 (De Gelder et al. 2005), pKJK10 (Musovic et al. 2006; Claudia et al.
2013), gfp-tagged IncP-1α plasmid RP4 (Musovic et al. 2010), pBP136, pCAR1,
NAH7 (Shintani et al. 2014), and low GC-type plasmid pHHV216 (Jechalke et al.
2013).
Soil microcosms utilized in gene transfer studies vary in complexity from simple
closed vessels containing a few grams of soil in a conical flask or falcon tube to
more complicated systems including vertical soil columns. Other microcosms
include continuous flow reactors through which water or nutrients are percolated.
Microcosms for studying bacterial interactions in the rhizosphere/rhizoplane were
elaborated by Hill and Top (1998). The microcosm system adopted depends upon
the objectives of the study. Care should be taken to assess the desired factors affect-
ing gene transfer in the soil/rhizosphere (Meena et al. 2013b, 2014b, 2015b, c, d;
Rawat et al. 2016; Yasin et al. 2016; Verma et al. 2015a; Shrivastava et al. 2016;
Velazquez et al. 2016; Sindhu et al. 2016; Singh et al. 2016). Important factors
include inoculum treatment and mode of application, concentration and sampling
methods, soil temperature, soil depth, concentrations of media, and types and
amounts of degradable chemicals (Hill and Top 1998; Wang et al. 2014a, b).
Table 6.1 shows the various soil factors affecting gene transfer studies in
microcosms.
6 Horizontal Gene Transfer in Soil and the Rhizosphere: Impact on Ecological… 121
6.6 Conclusions
The current state of understanding of HGT in soil and other natural environments
demonstrates that HGT is a mechanism of bacterial chromosomal evolution which
provides real-time adaptation among bacteria. HGT also provides genetic diversity
through its gene pool, which helps bacteria adapt to changing environmental condi-
tions. The availability of nutrients in the plant rhizosphere attracts bacteria, thus
offering a greater opportunity for HGT compared to bulk soil. Microcosm and bio-
film studies demonstrate that bacteria receive advantages and long-term ecological
benefits for survival and adaptation through HGT. However, the exact mechanisms
and magnitude of HGT in unsterile soil and in the rhizosphere must be explored
further, as factors influencing gene transfer in such situations are complex and
multifactorial.
Considering the untapped diversity of MGEs in soil- and plant-associated micro-
biomes, the impact of HGT on influencing plant-microbe interactions must be fur-
ther explored for possible exploitation in sustainable agriculture.
Acknowledgments We thank the editors and anonymous reviewers for their constructive com-
ments, which helped us to improve the manuscript.
122 M. Maheshwari et al.
References
Abulreesh HH (2011) Multidrug-resistant staphylococci in the environment. Int Conf Biotechnol
Environ Manag IPCBEE 18:1–6
Abulreesh HH (2012) Salmonellae in the environment. In: Annous BA, Gurtler JB (eds)
Salmonella-distribution, adaptation, control measures and technologies. Intech Open Access
Publisher, Rijeka., Chapter 2, pp 19–50
Ahmad M, Nadeem SM, Naveed M, Zahir ZA (2016) Potassium-solubilizing bacteria and their
application in agriculture. In: Meena VS, Maurya BR, Verma JP, Meena RS (eds) Potassium
solubilizing microorganisms for sustainable agriculture. Springer, New Delhi, pp 293–313.
doi:10.1007/978-81-322-2776-2_21
Aminov RI (2011) Horizontal gene exchange in environmental microbiota. Front Microbiol 2:158
Antonova ES, Hammer BK (2011) Quorum-sensing autoinducer molecules produced by members
of a multispecies biofilm promote horizontal gene transfer to Vibrio cholera. FEMS Microbiol
Lett 322(1):68–76
Arnold DL, Jackson RW, Waterfield NR, Mansfield JW (2007) Evolution of microbial virulence:
the benefits of stress. Trends Genet 23(6):293–300
Ashelford KE, Norris SJ, Fry JC, Bailey MJ, Day MJ (2000) Seasonal population dynamics and
interactions of competing bacteriophages and their host in the rhizosphere. Appl Environ
Microbiol 66(10):4193–4199
Ashelford KE, Day MJ, Fry JC (2003) Elevated abundance of bacteriophage infecting bacteria in
soil. Appl Environ Microbiol 69:285–289
Bahadur I, Meena VS, Kumar S (2014) Importance and application of potassic biofertilizer in
Indian agriculture. Int Res J Biol Sci 3:80–85
Bahadur I, Maurya BR, Kumar A, Meena VS, Raghuwanshi R (2016a) Towards the soil sustain-
ability and potassium-solubilizing microorganisms. In: Meena VS, Maurya BR, Verma JP,
Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture. Springer,
New Delhi, pp 225–266. doi:10.1007/978-81-322-2776-2_18
Bahadur I, Maurya BR, Meena VS, Saha M, Kumar A, Aeron A (2016b) Mineral release dynamics
of tricalcium phosphate and waste muscovite by mineral-solubilizing rhizobacteria isolated
from indo-gangetic plain of India. Geomicrobiol J. doi:10.1080/01490451.2016.1219431
Bais HP, Weir TL, Perry LG, Gilroy S, Vivanco JM (2006) The role of root exudates in rhizosphere
interactions with plants and other organisms. Annu Rev Plant Biol 57:233–266
Baltrus DA (2013) Exploring the costs of horizontal gene transfer. Trends Ecol Evol 28(8):489–495
Bardaji L, Pérez-Martinez I, Rodriguez-Moreno L, Rodríguez-Palenzuela P, Sundin GW, Ramos
C, Murillo J (2011) Sequence and role in virulence of the three plasmid complement of the
model tumor-inducing bacterium Pseudomonas savastanoi pv. savastanoi NCPPB 3335. PLoS
One 6:e25705
Barlow M (2009) What antimicrobial resistance has taught us about horizontal gene transfer.
Methods Mol Biol 532:397–411
Bolton H Jr, Fredrickson JK, Thomas JM, Shu-Mei WL, Workman DJ, Bentjen SA, Smith JL
(1991) Field calibration of soil-core microcosms: ecosystem structural and functional compari-
sons. Microb Ecol 21(1):175–189
Boon N, Goris J, De Vos P, Verstraete W, Top EM (2001) Genetic diversity among 3-chloroaniline-and
aniline-degrading strains of the Comamonadaceae. Appl Environ Microbiol 67(3):1107–1115
Boto L (2010) Horizontal gene transfer in evolution: facts and challenges. Proc Roy Soc B Biol
Soc 277(1683):819–827
Broszat M, Grohmann E (2014) Horizontal gene transfer in planktonic and biofilm modes. In:
Rumbaugh KP, Ahmad I (eds) Antibiofilm agents, Springer series on Biofilm 8. Springer,
Heidelberg, pp 125–162. ISBN: 978-3-642-53833-9
Brüssow H, Canchaya C, Hardt W (2004) Phages and the evolution of bacterial pathogens: from
genomic rearrangements to lysogenic conversion. Microbiol Mol Biol Rev 68(3):560–602
6 Horizontal Gene Transfer in Soil and the Rhizosphere: Impact on Ecological… 123
Burmølle M, Bahl MI, Jensen LB, Sørensen SJ, Hansen LH (2008) Type 3 fimbriae, encoded
by the conjugative plasmid pOLA52, enhance biofilm formation and transfer frequencies in
Enterobacteriaceae strains. Microbiology 154(1):187–195
Burmølle M, Ren D, Bjarnsholt T, Sørensen SJ (2014) Interactions in multispecies biofilms: do
they actually matter? Trends Microbiol 22(2):84–91
Burroughs NJ, Marsh P, Wellington EM (2000) Mathematical analysis of growth and interac-
tion dynamics of streptomycetes and a bacteriophage in soil. Appl Environ Microbiol
66(9):3868–3877
Cabezon E, Ripoll-Rozada J, Peña A, de la Cruz F, Arechaga I (2015) Towards an integrated model
of bacterial conjugation. FEMS Microbiol Rev 39(1):81–95
Canchaya C, Proux C, Fournous G, Bruttin A, Brüssow H (2003) Prophage genomics. Microbiol
Mol Biol Rev 67:238–276
Chen J, Carpena N, Quiles-Puchalt N, Ram G, Novick RP, Penadés JR (2015) Intra-and inter-
generic transfer of pathogenicity island-encoded virulence genes by cos phages. ISME
J9(5):1260–1263
Claudia I, Ren D, Blanchet M, Dendooven L, Marsch R, Sørensen SJ, Burmølle M (2013) The
ability of soil bacteria to receive the conjugative IncP1 plasmid, pKJK10, is different in a mixed
community compared to single strains. FEMS Microbiol Lett 338(1):95–100
Clegg S, Wilson J, Johnson J (2011) More than one way to control hair growth: regulatory mech-
anisms in enterobacteria that affect fimbriae assembled by the chaperone/usher pathway.
J Bacteriol 193(9):2081–2088
Conibear TC, Collins SL, Webb JS (2009) Role of mutation in Pseudomonas aeruginosa biofilm
development. PLoS One 4(7):6289
Cook LC, Dunny GM (2014) The influence of biofilms in the biology of plasmids. Microbiol
Spectr 2(5):12
Cordero OX, Wildschutte H, Kirkup B, Proehl S, Ngo L, Hussain F, Le Roux F, Mincer T, Polz MF
(2012) Ecological populations of bacteria act as socially cohesive units of antibiotic production
and resistance. Science 337(6099):1228–1123
Dahlberg C, Chao L (2003) Amelioration of the cost of conjugative plasmid carriage in Escherichia
coli K12. Genetics 165:1641–1649
Dalmasso M, Hill C, Ross RP (2014) Exploiting gut bacteriophages for human health. Trends
Microbiol 22(7):399–405
D’Alvise PW, Sjøholm OR, Yankelevich T, Jin Y, Wuertz S, Smets B (2010) TOL plasmid carriage
enhances biofilm formation and increases extracellular DNA content in Pseudomonas putida
KT2440. FEMS Microbiol Lett 312(1):84–92
Das I, Pradhan M (2016) Potassium-solubilizing microorganisms and their role in enhancing
soil fertility and health. In: Meena VS, Maurya BR, Verma JP, Meena RS (eds) Potassium
solubilizing microorganisms for sustainable agriculture. Springer, New Delhi, pp 281–291.
doi:10.1007/978-81-322-2776-2_20
Davies J, Davies D (2010) Origins and evolution of antibiotic resistance. Microbiol Mol Biol R
74(3):417–433
De Gelder L, Vandecasteele FP, Brown CJ, Forney LJ, Top EM (2005) Plasmid donor affects
host range of promiscuous IncP-1β plasmid pB10 in an activated-sludge microbial community.
Appl Environ Microbiol 71(9):5309–5317
de la Cueva-Méndez G, Pimentel B (2007) Gene and cell survival: lessons from prokaryotic plas-
mid R1. EMBO Rep 8(5):458–464
De Lipthay JR, Rasmussen LD, Oregaard G, Simonsen K, Bahl MI, Kroer N, Sørensen SJ
(2008) Acclimation of subsurface microbial communities to mercury. FEMS Microbiol Ecol
65(1):145–155
Dennis JJ (2005) The evolution of IncP catabolic plasmids. Curr Opin Biotechnol 16(3):291–298
Dominguez-Nunez JA, Benito B, Berrocal-Lobo M, Albanesi A (2016) Mycorrhizal fungi: role
in the solubilization of potassium. In: Meena VS, Maurya BR, Verma JP, Meena RS (eds)
Potassium solubilizing microorganisms for sustainable agriculture. Springer, New Delhi,
pp 77–98. doi:10.1007/978-81-322-2776-2_6
124 M. Maheshwari et al.
Dotaniya ML, Meena VD, Basak BB, Meena RS (2016) Potassium uptake by crops as well
as microorganisms. In: Meena VS, Maurya BR, Verma JP, Meena RS (eds) Potassium
solubilizing microorganisms for sustainable agriculture. Springer, New Delhi, pp 267–280.
doi:10.1007/978-81-322-2776-2_19
Dutta C, Sarkar M (2015) Horizontal gene transfer and bacterial diversity. In: Encyclopedia of
metagenomics. Springer, New York, pp 251–257
Etchuuya R, Ito M, Kitano S, Shigi F, Sobue R, Maeda S (2011) Cell-to-cell transformation in
Escherichia coli: a novel type of natural transformation involving cell-derived DNA and a puta-
tive promoting pheromone. PLoS One 6(1):16355
Frost LS, Leplae R, Summers AO, Toussaint A (2005) Mobile genetic elements: the agents of open
source evolution. Nat Rev Microbiol 3(9):722–732
Gardiner DM, Upadhyaya NM, Stiller J, Ellis JG, Dodds PN, Kazan K, Manners JM (2014)
Genomic analysis of Xanthomonas translucens pathogenic on wheat and barley reveals cross-
kingdom gene transfer events and diverse protein delivery systems. PLoS One 9(1):e84995
Ghigo JM (2001) Natural conjugative plasmids induce bacterial biofilm development. Nature
412(6845):442–445
Ghosh D, Roy K, Williamson KE, White DC, Wommack KE, Sublette KL, Radosevich M (2008)
Prevalence of lysogeny among soil bacteria and presence of 16S rRNA and trzN genes in viral-
community DNA. Appl Environ Microbiol 74(2):495–502
Gillings MR, Holley MP, Stokes HW, Holmes AJ (2005) Integrons in Xanthomonas: a source of
species genome diversity. Proc Natl Acad Sci USA 102:4419–1124
Gyles C, Boerlin P (2013) Horizontally transferred genetic elements and their role in pathogenesis
of bacterial disease. Vet Pathol 51(2):328–340
Hacker J, Kaper JB (2000) Pathogenicity islands and the evolution of microbes. Annu Rev
Microbiol 54:641–679
Hall-Stoodley L, Costerton JW, Stoodley P (2004) Bacterial biofilms: from the natural environ-
ment to infectious diseases. Nat Rev Microbiol 2:95–108
Heuer H, Smalla K (2007) Horizontal gene transfer between bacteria. Environ Biosaf Res
6(1–2):3–13
Heuer H, Smalla K (2012) Plasmids foster diversification and adaptation of bacterial populations
in soil. FEMS Microbiol Rev 36(6):1083–1104
Heuer H, Fox RE, Top EM (2007) Frequent conjugative transfer accelerates adaptation of a broad-
host-range plasmid to an unfavourable Pseudomonas putida host. FEMS Microbiol Ecol
59:738–748
Heuer H, Kopmann C, Binh CT, Top EM, Smalla K (2009) Spreading antibiotic resistance through
spread manure: characteristics of a novel plasmid type with low% G+ C content. Environ
Microbiol 11(4):937–949
Hill KE, Top EM (1998) Gene transfer in soil systems using microcosms. FEMS Microbiol Ecol
25(4):319–329
Huddleston JR (2014) Horizontal gene transfer in the human gastrointestinal tract: potential spread
of antibiotic resistant genes. Infect Drug Resist 7:167–176
Imfeld G, Vuilleumier S (2012) Measuring the effects of pesticides on bacterial communities in
soil: a critical review. Eur J Soil Biol 49:22–30
Jain R, Rivera MC, Moore JE, Lake JA (2003) Horizontal gene transfer accelerates genome inno-
vation and evolution. Mol Biol Evol 20(10):1598–1602
Jaiswal DK, Verma JP, Prakash S, Meena VS, Meena RS (2016) Potassium as an important plant
nutrient in sustainable agriculture: a state of the art. In: Meena VS, Maurya BR, Verma JP,
Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture. Springer,
New Delhi, pp 21–29. doi:10.1007/978-81-322-2776-2_2
Jat LK, Singh YV, Meena SK, Meena SK, Parihar M, Jatav HS, Meena RK, Meena VS (2015)
Does integrated nutrient management enhance agricultural productivity? J Pure Appl Microbiol
9(2):1211–1221
6 Horizontal Gene Transfer in Soil and the Rhizosphere: Impact on Ecological… 125
Merkey BV, Lardon LA, Seoane JM, Kreft JU, Smets BF (2011) Growth dependence of con-
jugation explains limited plasmid invasion in biofilms: an individual-based modelling study.
Environ Microbiol 13(9):2435–2452
Merlin C, Springael D, Toussaint A (1999) Tn4371: a modular structure encoding a phage-like
integrase, a pseudomonas-like catabolic pathway, and rp4/ti-like transfer functions. Plasmid
41(1):40–54
Molbak L, Søren M, Kroer N (2007) Root growth and exudate production define the frequency of
horizontal plasmid transfer in the rhizosphere. FEMS Microbiol Ecol 59(1):167–176
Molin S, Tolker-Nielsen T (2003) Gene transfer occurs with enhanced efficiency in biofilms and
induces enhanced stabilisation of the biofilm structure. Curr Opin Biotechnol 14(3):255–261
Musovic S, Oregaard G, Kroer N, Sørensen SJ (2006) Cultivation-independent examination of hor-
izontal transfer and host range of an IncP-1 plasmid among gram-positive and gram-negative
bacteria indigenous to the barley rhizosphere. Appl Environ Microbiol 72(10):6687–6692
Musovic S, Dechesne A, Sørensen J, Smets BF (2010) Novel assay to assess permissiveness of a
soil microbial community toward receipt of mobile genetic elements. Appl Environ Microbiol
76(14):4813–4818
Navarre WW (2016) The impact of gene silencing on horizontal gene transfer and bacterial evolu-
tion. Adv Microb Physiol 69:157–186
Nielsen KM, van Elsas JD (2001) Stimulatory effects of compounds present in the rhizosphere on
natural transformation of Acinetobacter sp. BD413 in soil. Soil Biol Biochem 33(3):345–357
Norman A, Hansen LH, Sørensen SJ (2009) Conjugative plasmids: vessels of the communal gene
pool. Philos Trans R Soc B 364(1527):2275–2289
Obeng N, Pratama AA, van Elsas JD (2016) The significance of mutualistic phages for bacterial
ecology and evolution. Trends Microbiol 24(6):440–449
Ochman H, Lawrence JG, Groisman EA (2000) Lateral gene transfer and the nature of bacterial
innovation. Nature 405(6784):299–304
Ojala V, Mattila S, Hoikkala V, Bamford JK, Jalasvuori M (2014) Evolutionary rescue of bacteria
via horizontal gene transfer under a lethal β-lactam concentration. J Glob Antimicrob Resist
2(3):198–200
Parewa HP, Yadav J, Rakshit A, Meena VS, Karthikeyan N (2014) Plant growth promoting rhizo-
bacteria enhance growth and nutrient uptake of crops. Agric Sustain Dev 2(2):101–116
Pearce SL, Oakeshott JG, Pandey G (2015) Insights into ongoing evolution of the hexachlorocy-
clohexane catabolic pathway from comparative genomics of ten Sphingomonadaceae strains.
G3 (Bethesda) 5(6):1081–1094
Petrova LP, Shelud’ko AV, Katsy EI (2010) Plasmid rearrangements and alterations in Azospirillum
brasilense biofilm formation. Microbiology 79(1):121–124
Pinto-Carbo M, Sieber S, Dessein S, Wicker T, Verstraete B, Gademann K, Eberl L, Carlier A
(2016) Evidence of horizontal gene transfer between obligate leaf nodule symbionts. ISME
J 10:2092–2105
Powell BJ, Purdy KJ, Thompson IP, Bailey MJ (1993) Demonstration of tra+ plasmid activity in
bacteria indigenous to the phyllosphere of sugar beet; gene transfer to a recombinant pseudo-
monad. FEMS Microbiol Ecol 12:195–206
Prakash S, Verma JP (2016) Global perspective of potash for fertilizer production. In: Meena VS,
Maurya BR, Verma JP, Meena RS (eds) Potassium solubilizing microorganisms for sustainable
agriculture. Springer, New Delhi, pp 327–331. doi:10.1007/978-81-322-2776-2_23
Priyadharsini P, Muthukumar T (2016) Interactions between arbuscular mycorrhizal fungi and
potassium-solubilizing microorganisms on agricultural productivity. In: Meena VS, Maurya
BR, Verma JP, Meena RS (eds) Potassium solubilizing microorganisms for sustainable agricul-
ture. Springer, New Delhi, pp 111–125. doi:10.1007/978-81-322-2776-2_8
Raghavendra MP, Nayaka NC, Nuthan BR (2016) Role of rhizosphere microflora in potas-
sium solubilization. In: Meena VS, Maurya BR, Verma JP, Meena RS (eds) Potassium
solubilizing microorganisms for sustainable agriculture. Springer, New Delhi, pp 43–59.
doi:10.1007/978-81-322-2776-2_4
128 M. Maheshwari et al.
Rankin DJ, Rocha EP, Brown SP (2011) What traits are carried on mobile genetic elements, and
why & quest. Heredity 106(1):1–10
Rawat J, Sanwal P, Saxena J (2016) Potassium and its role in sustainable agriculture. In: Meena
VS, Maurya BR, Verma JP, Meena RS (eds) Potassium solubilizing microorganisms for sus-
tainable agriculture. Springer, New Delhi, pp 235–253. doi:10.1007/978-81-322-2776-2_17
Reisner A, Höller BM, Molin S, Zechner EL (2006) Synergistic effects in mixed Escherichia coli
biofilms: conjugative plasmid transfer drives biofilm expansion. J Bacteriol 188(10):3582–3588
Røder HL, Hansen LH, Sørensen SJ, Burmølle M (2013) The impact of the conjugative IncP-1
plasmid pKJK5 on multispecies biofilm formation is dependent on the plasmid host. FEMS
Microbiol Lett 344(2):186–192
Russell RJ, Scott C, Jackson CJ, Pandey R, Pandey G, Taylor MC, Coppin CW, Liu JW, Oakeshott
JG (2011) The evolution of new enzyme function: lessons from xenobiotic metabolizing bacte-
ria versus insecticide-resistant insects. Evol Appl 4(2):225–248
Saha M, Maurya BR, Bahadur I, Kumar A, Meena VS (2016a) Can potassium-solubilising bacteria
mitigate the potassium problems in India? In: Meena VS, Maurya BR, Verma JP, Meena RS
(eds) Potassium solubilizing microorganisms for sustainable agriculture. Springer, New Delhi,
pp 127–136. doi:10.1007/978-81-322-2776-2_9
Saha M, Maurya BR, Meena VS, Bahadur I, Kumar A (2016b) Identification and characterization
of potassium solubilizing bacteria (KSB) from Indo-Gangetic Plains of India. Biocatal Agric
Biotechnol 7:202–209
Sangwan N, Verma H, Kumar R, Negi V, Lax S, Khurana P, Khurana JP, Gilbert JA, Lal R (2014)
Reconstructing an ancestral genotype of two hexachlorocyclohexane-degrading Sphingobium
species using metagenomic sequence data. ISME J 8(2):398–408
Schlüter A, Szczepanowski R, Pühler A, Top EM (2007) Genomics of IncP-1 antibiotic resistance
plasmids isolated from wastewater treatment plants provides evidence for a widely accessible
drug resistance gene pool. FEMS Microbiol Rev 31(4):449–477
Sharma A, Shankhdhar D, Shankhdhar SC (2016) Potassium-solubilizing microorganisms:
mechanism and their role in potassium solubilization and uptake. In: Meena VS, Maurya BR,
Verma JP, Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture.
Springer, New Delhi, pp 203–219. doi:10.1007/978-81-322-2776-2_15
Shintani M, Matsui K, Inoue JI, Hosoyama A, Ohji S, Yamazoe A, Nojiri H, Kimbara K, Ohkuma
M (2014) Single-cell analyses revealed transfer ranges of IncP-1, IncP-7, and IncP-9 plasmids
in a soil bacterial community. Appl Environ Microbiol 80(1):138–145
Shrivastava M, Srivastava PC, D’Souza SF (2016) KSM soil diversity and mineral solubilization,
in relation to crop production and molecular mechanism. In: Meena VS, Maurya BR, Verma JP,
Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture. Springer,
New Delhi, pp 221–234. doi:10.1007/978-81-322-2776-2_16
Sindhu SS, Parmar P, Phour M, Sehrawat A (2016) Potassium-solubilizing microorganisms
(KSMs) and its effect on plant growth improvement. In: Meena VS, Maurya BR, Verma JP,
Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture. Springer,
New Delhi, pp 171–185. doi:10.1007/978-81-322-2776-2_13
Singh NP, Singh RK, Meena VS, Meena RK (2015) Can we use maize (Zea mays) rhizobacteria as
plant growth promoter? Vegetos 28(1):86–99. doi:10.5958/2229-4473.2015.00012.9
Singh M, Dotaniya ML, Mishra A, Dotaniya CK, Regar KL, Lata M (2016) Role of biofertil-
izers in conservation agriculture. In: Bisht JK, Meena VS, Mishra PK, Pattanayak A (eds)
Conservation agriculture: an approach to combat climate change in Indian Himalaya. Springer,
Singapore, pp 113–134. doi:10.1007/978-981-10-2558-7_4
Smalla K, Borin S, Heuer H, Gebhard F, van Elsas JD, Nielsen K (2000) Horizontal transfer
of antibiotic resistance genes from transgenic plants to bacteria. In: Fairbairn C, Scoles G,
McHugen A (eds) The biosafety of genetically modified organisms. University Extension
Press, University of Saskatchewan
Smets BF, Barkay T (2005) Horizontal gene transfer: perspectives at a crossroads of scientific
disciplines. Nat Rev Microbiol 3(9):675–678
6 Horizontal Gene Transfer in Soil and the Rhizosphere: Impact on Ecological… 129
Sørensen SJ, Bailey M, Hansen LH, Kroer N, Wuertz S (2005) Studying plasmid horizontal trans-
fer in situ: a critical review. Nat Rev Microbiol 3(9):700–710
Springael D, Top EM (2004) Horizontal gene transfer and microbial adaptation to xenobiotics:
new types of mobile genetic elements and lessons from ecological studies. Trends Microbiol
12(2):53–58
Stalder T, Top E (2016) Plasmid transfer in biofilms: a perspective on limitations and opportuni-
ties. NPJ Biofilms Microbiomes 2:16022
Stewart PS, Franklin MJ (2008) Physiological heterogeneity in biofilms. Nat Rev Microbiol
6(3):199–210
Sun B, Li T, Xiao J, Liu L, Zhang P, Murphy RW, He S, Huang D (2016) Contribution of multiple-
inter-kingdom horizontal gene transfers to evolution and adaptation of amphibian-killing
Chytrid, Batrachochytrium dendrobatidis. Front Microbiol 7:1360
Teotia P, Kumar V, Kumar M, Shrivastava N, Varma A (2016) Rhizosphere microbes: potassium
solubilization and crop productivity-present and future aspects. In: Meena VS, Maurya BR,
Verma JP, Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture.
Springer, New Delhi, pp 315–325. doi:10.1007/978-81-322-2776-2_22
Toleman MA, Bennett PM, Walsh TR (2006) Common regions eg orf513 and antibiotic resistance:
IS91-like elements evolving complex class 1 integrons. J Antimicrob Chemother 58(1):1–6
Top EM, Springael D (2003) The role of mobile genetic elements in bacterial adaptation to xeno-
biotic organic compounds. Curr Opin Biotechnol 14(3):262–269
Tormo MA, Knecht E, Götz F, Lasa I, Penades JR (2005) Bap-dependent biofilm formation by
pathogenic species of Staphylococcus: evidence of horizontal gene transfer? Microbiology
151(7):2465–2475
van Elsas JD, Bailey MJ (2002) The ecology of transfer of mobile genetic elements. FEMS
Microbiol Ecol 42(2):187–197
van Elsas JD, Turner S, Bailey MJ (2003) Horizontal gene transfer in the phytosphere. New Phytol
157(3):525–537
van Overbeek LS, Wellington EMH, Egan S, Smalla K, Heuer H, Collard JM, Guillaume G,
Karagouni AD, Nikolakopoulou TL, van Elsas JD (2002) Prevalence of streptomycin resis-
tance genes in bacterial populations in European habitats. FEMS Microbiol Ecol 42:277–288
Velazquez E, Silva LR, Ramírez-Bahena MH, Peix A (2016) Diversity of potassium-solubilizing
microorganisms and their interactions with plants. In: Meena VS, Maurya BR, Verma JP,
Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture. Springer,
New Delhi, pp 99–110. doi:10.1007/978-81-322-2776-2_7
Verma R, Maurya BR, Meena VS (2014) Integrated effect of bio-organics with chemical fertilizer
on growth, yield and quality of cabbage (Brassica oleracea var capitata). Indian J Agric Sci
84(8):914–919
Verma JP, Jaiswa DK, Meena VS, Meena RS (2015a) Current need of organic farming for enhanc-
ing sustainable agriculture. J Clean Prod 102:545–547
Verma JP, Jaiswal DK, Meena VS, Kumar A, Meena RS (2015b) Issues and challenges about sus-
tainable agriculture production for management of natural resources to sustain soil fertility and
health. J Clean Prod 107:793–794
Wang Y, Xiao M, Geng X, Liu J, Chen J (2007) Horizontal transfer of genetic determinants for
degradation of phenol between the bacteria living in plant and its rhizosphere. Appl Microbiol
Biotechnol 77(3):733–739
Wang Y, Kou S, Jiang Q, Xu B, Liu X, Xiao J, Tian Y, Zhou C, Zhang C, Xiao M (2014a) Factors
affecting transfer of degradative plasmids between bacteria in soils. Appl Soil Ecol 84:254–261
Wang Y, Jiang Q, Zhou C, Chen B, Zhao W, Song J, Fang R, Chen J, Xiao M (2014b) In-situ
remediation of contaminated farmland by horizontal transfer of degradative plasmids among
rhizosphere bacteria. Soil Use Manag 30(2):303–309
Watanabe T (1963) Infective heredity of multiple drug resistance in bacteria. Bacteriol Rev
27(1):87
Weitz JS, Poisot T, Meyer JR, Flores CO, Valverde S, Sullivan MB, Hochberg ME (2013) Phage–
bacteria infection networks. Trends Microbiol 21(2):82–91
130 M. Maheshwari et al.
Wellington EM, Boxall AB, Cross P, Feil EJ, Gaze WH, Hawkey PM, Johnson-Rollings AS, Jones
DL, Lee NM, Otten W, Thomas CM (2013) The role of the natural environment in the emer-
gence of antibiotic resistance in gram-negative bacteria. Lancet Infect Dis 13(2):155–165
Wiedenbeck J, Cohan FM (2011) Origins of bacterial diversity through horizontal genetic transfer
and adaptation to new ecological niches. FEMS Microbiol Rev 35(5):957–976
Williamson KE, Radosevich M, Smith DW, Wommack KE (2007) Incidence of lysogeny within
temperate and extreme soil environments. Environ Microbiol 9(10):2563–2574
Wommack KE, Colwell RR (2000) Virioplankton: viruses in aquatic ecosystems. Microbiol Mol
Biol Rev 64:69–114
Yadav BK, Sidhu AS (2016) Dynamics of potassium and their bioavailability for plant nutrition. In:
Meena VS, Maurya BR, Verma JP, Meena RS (eds) Potassium solubilizing microorganisms for
sustainable agriculture. Springer, New Delhi, pp 187–201. doi:10.1007/978-81-322-2776-2_14
Yasin M, Munir I, Faisal M (2016) Can Bacillus spp. enhance K+ uptake in crop species. In:
Meena VS, Maurya BR, Verma JP, Meena RS (eds) Potassium solubilizing microorganisms for
sustainable agriculture. Springer, New Delhi, pp 163–170. doi:10.1007/978-81-322-2776-2_12
Yuan J, Zhang N, Huang Q, Raza W, Li R, Vivanco JM, Shen Q (2015) Organic acids from root
exudates of banana help root colonization of PGPR strain Bacillus amyloliquefaciens NJN-6.
Sci Rep 5:13438
Zahedi H (2016) Growth-promoting effect of potassium-solubilizing microorganisms on
some crop species. In: Meena VS, Maurya BR, Verma JP, Meena RS (eds) Potassium sol-
ubilizing microorganisms for sustainable agriculture. Springer, New Delhi, pp 31–42.
doi:10.1007/978-81-322-2776-2_3
Zechner EL, de la Cruz F, Eisenbrandt R, Grahn AM, Koraimann G, Lanka E, Muth G, Pansegrau
W, Thomas CM, Wilkins BM, Zatyka M (2000) Conjugative-DNA transfer processes. In:
Thomas CM (ed) The horizontal genepool: bacterial plasmids and gene spread. Harwood
ScientificPublication, Amsterdam, pp 87–174
Zhang N, Wang D, Liu Y, Li S, Shen Q, Zhang R (2014) Effects of different plant root exudates and
their organic acid components on chemotaxis, biofilm formation and colonization by beneficial
rhizosphere-associated bacterial strains. Plant Soil 374(1–2):689–700
Zhao Y, Ma Z, Sundin GW (2005) Comparative genomic analysis of the pPT23A plasmid family
of Pseudomonas syringae. J Bacteriol 187:2113–2126
Hairy Root Culture: A Biotechnological
Approach to Produce Valuable 7
Metabolites
Hamideh Vaghari, Hoda Jafarizadeh-Malmiri,
Navideh Anarjan, and Aydin Berenjian
Abstract
Hairy root (HR) cultures are attracting more attention due to their unique ability
in degrading different pollutants and production of metabolites with therapeutic
or industrial applications. This specific type of plant cell culture is derived from
explants that are infected by Agrobacterium rhizogenes. The HR cultures
are categorized by their growth rate as well as their genetic and biochemical
stability. Progress in design of innovative bioreactors and process intensification
for HR growth will allow successful industrial production of metabolites. This
chapter will present advances in work on HR cultures related to the detoxifica-
tion of pollutants, production of valuable metabolites, and their cultivation in
large-scale intensified bioreactors.
Keywords
Hairy root • Metabolite • Scale up • Bioreactor • Process intensification
7.1 Introduction
Plants are able to produce a wide range of primary and secondary metabolites
(Yazaki et al. 2008; Sharma et al. 2013). However, the large-scale production of
these valuable compounds has been limited by low growth rates, climate depen-
dency, restricted cultivation areas, plant diseases, pests, overharvesting, and intense
labor requirement (Sharma et al. 2013; Meena et al. 2013a; Bahadur et al. 2014;
Maurya et al. 2014; Jat et al. 2015; Kumar et al. 2016b; Rates 2001). Moreover, the
chemical synthesis of plant-derived metabolites is not a feasible choice due to their
complex structures and their specific stereochemical requirements (Sharma et al.
2013; Namdeo 2007). All these issues emphasize the need for developing new
methods and protocols for the industrial-level fabrication of plant-derived
metabolites.
Suspension culture of plant cells has been considered as another promising
source for biosynthesis of valuable secondary metabolites (Sharma et al. 2013).
More than 25% of the available pharmaceuticals are either based on originally
found compounds in plants or are extracted from them (Giri and Narasu 2000).
Production of secondary metabolites by using suspended plant cell culture is usu-
ally a challenging task as these compounds produce at distinct developmental
stages. Therefore, in vitro studies of differentiated and organized tissues (mainly the
roots) have been developed and were reported to be a more predictable approach as
compared to cell suspension cultures (Sharma et al. 2013; Kumar et al. 2015; Ahmad
et al. 2016; Meena et al. 2016a; Parewa et al. 2014).
The plant roots are suitable for large-scale production since they are the key
point for synthesis and/or storage of certain chemicals. The biotechnological fabri-
cation of wide range of valuable secondary metabolites by using plant cultures can
be seen as an alternative to the extraction of whole plant material (Namdeo 2007).
Several strategies have been investigated in order to further enhance the production
of secondary metabolites from medicinal plants. Some of these include high yield-
ing cell line screening, media modification, elicitation, precursor feeding, large-
scale cultivation system, plant cell immobilization, hairy root culture,
biotransformation, and others (Rao and Ravishankar 2002; Vanisree et al. 2004).
Recently, hairy root (HR) culture has been developed in order to inhibit the use
of large volumes of plants that are needed to be purified. Totipotency is among the
major characteristics of plant cells; therefore, HRs could successfully produce pri-
mary and secondary metabolites similar to intact roots (Giri and Narasu 2000;
Qaderi et al. 2016). HR culture is a tool that makes use of soil bacterium
Agrobacterium rhizogenes ability to transfer genes to the genome of the host plant
(Sharma et al. 2013; Thwe et al. 2016). This technique was developed as the innova-
tive path for bulky production of secondary metabolite and phytochemicals which
allows developing large amount of roots and secondary metabolites in short time for
continuous supply of improved value products (Korde et al. 2016). These HRs have
also been used for root physiology and biosynthetic pathway (Giri and Narasu
2000), regeneration of whole plants with desirable phenotypes, and phytoremedia-
tion of toxic substances and reactive dyes (Talano et al. 2012; Prakash and Verma
7 Hairy Root Culture: A Biotechnological Approach to Produce Valuable Metabolites 133
2016; Meena et al. 2015a, 2016b; Priyadharsini and Muthukumar 2016; Kumar
et al. 2017). Finally, HR cultures include other aspects such as molecular metabolic
engineering, bioreactor design, and optimization (Ono and Tian 2011). In this chap-
ter, we present advances in work on HR cultures related to the detoxification of
pollutants, production of valuable metabolites, and fabrication in large-scale inten-
sified bioreactors. A suggestion to overcome current challenges and emerging trends
for future progression of research has also been provided.
HR production is carried out through the plant tissue culture technique in order to
study the plant metabolic processes or to manufacture precious secondary metabo-
lites with the use of plant genetic engineering. HR culture is also called as trans-
formed root culture from gram-negative soil bacterium A. rhizogenes that contains
root-inducing plasmids (Ri plasmids) (Korde et al. 2016; Pistelli et al. 2010). It
infects roots of dicot, and some monocot plants cause them to produce the opines
which is a type of unusual amino acids (octopine, agropine, nopaline, mannopine,
and cucumopine). Such opines are used by the bacterium as a carbon, nitrogen, and
energy source (Ferdosi and Kashefi 2014).
The morphology of HRs is significantly different from the normal roots as they
are much more branched and have much lateral meristematic growth, which will
lead to higher biomass. The abnormal roots however are easier to grow in artificial
media without hormone, and they are neoplastic in nature, with hazy growth.
Fabricated HRs by infection of A. rhizogenes have a high growth rate as well as
genetic and biochemical makeup (Korde et al. 2016).
However, new techniques are developed in order to make HRs by the use of new
plant species (Georgiev et al. 2011). HRs have numerous advantages such as indefi-
nite and fast in vitro growth even in the absence of phytohormones as well as high
genotype and phenotype stability (Ono and Tian 2011). HR culture is among the
main cultures that are used to investigate metabolic processes of plants, secondary
metabolites production, recombinant proteins, plant genetic engineering, phytore-
mediation, artificial seed production, biofortification, and biopharmaceuticals.
Applications of these efficient technologies also include several aspects as meta-
bolic engineering, bioreactor design, and process optimization (Raghavendra et al.
2016; Zahedi 2016; Meena et al. 2015b; Rawat et al. 2016; Kumar et al. 2016a).
Ti Plasmid
Opine
Catabolism
Virulence
Region Origin of
Replication
(ORI)
auxins and the synthesis of mannopine and agropine. After the transfer of the TL-
DNA and TR-DNA, they are integrated in the genome of the plant cell. The TL-DNA
is vital for the hairy root induction (Chandra 2012).
Successful HR culture system requires several essential check marks, namely, selec-
tion of best A. rhizogenes strain, appropriate explants and antibiotic, and a suitable
culture medium (Sharma et al. 2013). Strains of A. rhizogenes are widely varying in
their transforming ability. HRs that are fabricated by using different types of bacte-
rial strains show significantly different morphologies. These observed virulence and
morphology differences could be justified by the different strain plasmid harbored
(Saha et al. 2016a; Yadav and Sidhu 2016; Nguyen et al. 1992; Meena et al. 2015f).
Most plant materials like hypocotyl, stem, cotyledon, leaf, tuber, or storage root
may be applied to make HRs (Królicka et al. 2001; Sevón and Oksman-Caldentey
2002; Giri et al. 2001). In order to induce HRs, explants should be infected with
strains of A. rhizogenes either by cocultivation or direct inoculation (Giri et al.
2001; Ur Rahman et al. 2004). Subsequently, roots are subculture using a medium
such as MS or B5 (Fig. 7.2) (Sevón and Oksman-Caldentey 2002; Le Flem-
Bonhomme et al. 2004; Palazón et al. 2003a, b).
The HR culture shaves diverse and abundant applications (Fig. 7.3). They tradition-
ally have been used to investigate root physiology in conjunction with biosynthetic
pathway elucidation (Ibanez et al. 2016). Nowadays, HR culture technique is now
being used for the fabrication of bioactive compounds, secondary metabolites, and
phytochemicals. HRs are popular for regeneration of whole plants with desirable
phenotypes by infection of ornamental plants with A. rhizogenes (Meena et al.
2016d; Saha et al. 2016b; Verma et al. 2015b; Bahadur et al. 2016b; Das and Pradhan
2016; Dominguez-Nunez et al. 2016; Dotaniya et al. 2016).
Moreover, HR cultures have been used for phytoremediation of toxic substances
and reactive dye. Presently, several high-value bioactives are fabricated by using
HRs from various plant sources which have application in pharmaceutical and cos-
metic products (Ono and Tian 2011). Furthermore, recombinant protein production
using this system was found to be a sustainable method for producing cytokines as
well as protein therapeutics (Talano et al. 2012).
136 H. Vaghari et al.
7.5.1 A
pplication of HR Cultures for Secondary Metabolites
Production
Plants are known as chemical factories since they pose the ability to fabricate impor-
tant phytochemical. However, the major issue is that their growth is dependent to
the outside environmental stress. HR cultures are promising source for phytochemi-
cal due to sizable biomass production and biosynthetic capacity.
Furthermore, HR cultures frequently accumulate phytochemical at much higher
levels than cell or callus cultures (McCoy and O’Connor 2008; Ono and Tian 2011).
Secondary metabolites as summarized in Table 7.1 are naturally more complex as
compared to primary metabolites. These compounds have been categorized into
terpenoids, phenolics, and alkaloids (Chinou 2008).
Several HR cultures have attracted significant amount of attention due to their
potential in production of valuable phytochemical including Artemisia annua,
Catharanthus roseus, Arachis hypogaea, and Camptotheca acuminata/Ophiorrhiza
pumila (McCoy and O’Connor 2008). List of secondary metabolites produced by
wild HR cultures is summarized in Table 7.2. Although, non-transgenic HR cultures
continue to serve as a good source for phytochemicals and secondary metabolites
(Ono and Tian 2011).
However, application of metabolic engineering methods requires acritical under-
standing about the regulation of secondary metabolite pathways and the metabolic
138 H. Vaghari et al.
Table 7.2 List of the secondary metabolites that are produced by wild HR cultures
Secondary metabolite Function HR References
Ajmalicine, ajmaline Antihypertensive Rauvolfia micrantha Sudha et al.
(2003)
Artemisinin Antimalarial Artemisia annua Weathers et al.
(2005)
Azadirachtin Biopesticide Azadirachta indica Srivastava and
Srivastava
(2012c)
Benzylisoquinoline Analgesic, antibiotic Papaver somniferum Park and
alkaloids (morphinan, Facchini (2000)
codeine, and and Le
sanguinarine) Bonhomme
et al. (2004)
Betalain Red pigments for food Beta vulgaris Rudrappa et al.
industry, strong (2004) and
aphrodisiac, laxative Pavlov et al.
(2003)
Camptothecin Antitumor, AIDS, Ophiorrhiza alata Ya-ut et al.
falciparum malaria, Craib, Ophiorrhiza (2011) and
colorectal and ovarian pumila Sato et al.
cancers treatment (2001)
3,4-Dihydroxyl-L- Therapeutic agent Stizolobium hassjoo Sung and
phenylalanine against Parkinson’s Huang (2006)
disease
Dopa and dopamine Neurotransmitters Beta vulgaris Rudrappa et al.
(2004)
Flavone glycosides Anti-inflammatory Catharanthus roseus Talano et al.
action (2012)
Flavonoids Meant for the treatment Glycyrrhiza Li et al. (2002)
of gastric ulcers, anti- pallidiflora
inflammatory, and
antitussive
Flavonoids Antimutagenic, Glycyrrhiza uralensis Zhang et al.
antiulcer, antitumor, (2009)
antimicrobial
Glycyrrhizin Artificial sweetener and Glycyrrhiza inflate Wongwicha
pharmaceutical products et al. (2011)
(peptic ulcers treatment)
Glycyrrhizin Diuretic, tonic, Abrus precatorius Dixit and
alexiteric, antifertility Vaidya (2010)
Hyoscyamine Narcotic and Datura stramonium Pavlov et al.
antispasmodic activity, (2009)
used against Parkinson’s
disease
Indole alkaloids Anticancer Catharanthus roseus Ayora-Talavera
(vinblastine, vincristine) et al. (2002)
Iridoid glycosides Anti-inflammatory, Harpagophytum Georgiev et al.
analgesic, antidiabetic procumbens (2006)
(continued)
140 H. Vaghari et al.
Table 7.2 (continued)
Secondary metabolite Function HR References
6-Methoxy- Anticancer Linum album, Linum Wink et al.
podophyllotoxin persicum (2005)
Physalins Diuretic, febrifuge, Physalis minima Azlan et al.
vermifuge (2002)
Plumbagin Diuretic, antibacterial Plumbago zeylanica Sivanesan and
and used against leprosy Jeong (2009)
Resveratrol Anti-inflammatory, Arachis hypogaea Kim et al.
antioxidant, anti- (2008)
infective, anticancer
Rosmarinic acid Astringent, antioxidant, Nepeta cataria Yang (2010)
anti-inflammatory,
antimutagenic,
antimicrobial, antiviral
Rutin Antioxidant, Fagopyrum Kim et al.
anticarcinogenic, esculentum (2010)
antithrombotic,
cytoprotective,
vasoprotective
Rutin, hispidulin, and Anti-inflammatory; Saussurea involucrata Fu et al. (2005)
syringin antifungal
Serpentine Diabetes treatment Catharanthus roseus Datta et al.
(2010)
Sesquiterpenes Phytoalexins Hyoscyamus albus Kawauchi et al.
(2010)
Shikonin Dye for silk and food Arnebia Talano et al.
industry, anti- (2012)
inflammatory, anti-
allergic, and
antineoplasic activities
Stilbenoids (resveratrol, Antioxidant, anticancer, Arachis hypogaea Medina-Bolivar
pinosylvin, and antiatherosclerosis, et al. (2010)
derivatives) neuroprotective, and
estrogenic activities
Tropane alkaloids Narcotic, anticholinergic Datura metel, Moyano et al.
and antispasmodic Hyoscyamus muticus (2003)
activity
Tropane alkaloids Narcotic, anticholinergic Hyoscyamus niger Zhang et al.
and antispasmodic (2004)
activity
Tropane alkaloids Narcotic, anticholinergic Datura innoxia Dechaux and
(scopolamine and and antispasmodic Boitel-Conti
hyoscyamine) activity (2005)
Tropane alkaloid Used against Parkinson’s Atropa belladonna Richter et al.
(hyoscyamine, atropine, disease (2005)
and hyoscine)
(continued)
7 Hairy Root Culture: A Biotechnological Approach to Produce Valuable Metabolites 141
Table 7.2 (continued)
Secondary metabolite Function HR References
Tropane alkaloids Parasympatholytic Przewalskia tangutica Lan and Quan
(scopolamine and (2010)
hyoscyamine)
Withanolide A Brain regenerative Withania somnifera Praveen and
properties Murthy (2012)
Xanthotoxin Leucoderma Ammi majus Krolicka et al.
(furocoumarin) (2001)
Root tissues are not identical to microbial cultures in many ways. Therefore, biore-
actors for HR cultures are more challenging to be controlled, operated, and scaled
up. Development of innovative bioreactors and process intensification will allow to
optimize cell growth and large-scale production (Sharma et al. 2016; Meena et al.
2013c, 2016e; Verma et al. 2015a; Bahadur et al. 2016a; Masood and Bano 2016).
7.6.1 D
evelopment of Groundbreaking Bioreactors in HR
Cultures
Design and optimization of bioreactors have been the great advance in HRs for
industrial-scale production of metabolites (Huang and McDonald 2012). Production
of HRs in bioreactors helps to have a better control on operating conditions and
consequently optimize the growth and biosynthesis of the secondary metabolite
(Eibl and Eibl 2008). Bioreactor optimization for fabrication of HRs is of critical
importance for scale-up strategies.
HR bioreactors can be in general divided into gas or liquid phase. In liquid-phase
bioreactors, roots are always placed in the medium; as a result they are called sub-
merged reactors. On the other hand, in gas-phase reactors, the roots are almost exposed
142 H. Vaghari et al.
Table 7.3 List of secondary metabolites produced through transgenic HR cultures (Talano et al.
2012)
Secondary
metabolite Function Transgenic HR Foreign genes
Solanoside Antineoplastic agent Solanum Gene encoding a
khasianum specific antibody that
binds solanoside
Indole Beneficial effects on cancer, Catharanthus Modified anthranilate
sedative and hypotensive roseus synthase (AS) alpha
action subunit (trp5) and
tryptophan
decarboxylase gene
(TDC)
Ginseng Traditional Chinese Panax ginseng cs gene for cycloartenol
medicine, tonic, antiaging, synthase enzyme
anticancer, and anti-
diabetes properties
Scopolamine Hyoscyamus Putrescine
niger N-methyltransferase
pmt) and hyoscyamine
6ß-hydroxylase (h6h)
genes
Catharanthine Anticholinergic agents that Catharanthus Geraniol
act on parasympathetic roseus 10-hydroxylase (G10H)
nervous system and a jasmonate-
responsive transcript
factor (ORCA3)
Hyoscyamine, Scopolia Putrescine
scopolamine parviflora N-methyltransferase
(pmt) and hyoscyamine
6ß- hydroxylase (h6h)
Anisodamine, Anisodus Putrescine
anisodine, acutangulus N-methyltransferase
hyoscyamine, (pmt) and gene
scopolamine codifying tropinone
reductase I (TRI)
Glycyrrhizin Medicine, healthcare Glycyrrhiza Chalcone synthase
products, food (sweetener), uralensis
and cosmetics
Flavones: Diuretic, anti-inflammatory, Scutellaria Chalcone isomerase
baicalin, antiseptic, antispasmodic, baicalensis
baicalein, and anticancer
wogonin
Vitamin C Antioxidant properties Solanum gal UR gene
lycopersicon
Total sterols Hypocholesterolemic, Centella asiatica Farnesyl diphosphate
anticarcinogenic properties synthase from Panax
ginseng
7 Hairy Root Culture: A Biotechnological Approach to Produce Valuable Metabolites 143
Fig. 7.4 Types of phytoremediation. Plant metabolizes the pollutant via organic and inorganic
phytoremediation or rhizospheric metabolism. Phytoremediation includes several approaches,
namely, phytostabilization, phytoextraction, phytodegradation, and phytostimulation
to air or another gas mixture (Kim et al. 2002a, b; Stiles and Liu 2013). The design of
the reactor also depends on the product location, which is either intracellular or extra-
cellular (Meena et al. 2013b, 2015d; Shrivastava et al. 2016; Singh et al. 2015).
HR cultivation is usually associated with clumps formation that are naturally
composed of primary roots and their bridged lateral roots. It is very difficult to find
appropriate bioreactor for HR cultures because the rheological properties of HR
cultures vary from one species to another and even within clones of a single species.
Several bioreactor designs have been reported for HRs (Mishra and Ranjan 2008).
Schematic diagrams of promising bioreactor types that have been successfully
tested are depicted in Fig. 7.5.
Table 7.4 (continued)
Chemical
nature of
pollutant Nature of HR Pollutant Plant species References
Reactive red 198 Tagetes patula L. Patil et al. (2009)
dye
Tetracycline, Helianthus annuus Gujarathi et al.
oxytetracycline (2005)
Textile dye: Methyl Brassica juncea. Telke et al. (2011)
orange
Textile dye: reactive Sesuvium Lokhande et al.
green 19A- HE4BD portulacastrum L. (2015)
Transgenic Phenol Nicotiana tabacum Alderete et al.
(2009)
Brassica juncea Gonzalez et al.
inoculated with two (2013)
rhizobacteria
Solanum Oller et al. (2005)
lycopersicum
Brassica juncea Ontañon et al.
inoculated with (2014)
Pantoea sp. FC1
Nicotiana tabacum Sosa Alderete
expressing tpx1 et al. (2009),
and/or tpx2 genes (2012)
Nicotiana tabacum Ibanez et al.
expressing tpx1 (2011)
gene + AMF
TCE Atropa belladonna Banerjee et al.
(2002)
DNT 2,4-dinitrotoluene, PCB polychlorinated biphenyls, TNT 2,4,6-trinitrotoluene, DANTs diami-
nonitrotoluenes, ADNTs aminodinitrotoluenes, AMF Arbuscular Mycorrhizal Fungi, TCE
trichloroethylene
Cooling jacket
Roots
Baffle Sparger Wire mesh
Roots
Wire mesh Agitator
Plastic liner
Roots
Sparger Sparger
Medium Air in
Air ut Mist
Air out Medium
Air in
Air in
Mist
Culture chamber
Roots
Roots
Sparger
Mist generator
Reservoir
Bubble column reactors (BCR) are among the simplest bioreactors that are easy
to scale up. The use of bubbles instead of mechanical mixers minimizes the shear
stress on the cultures (Choi et al. 2008; Huang and McDonald 2012). However, the
major drawback with BCRs are the undefined flow pattern of the liquid (Choi et al.
2008) and the reduced growth performance (Kwok and Doran 1995). In the
7 Hairy Root Culture: A Biotechnological Approach to Produce Valuable Metabolites 147
presence of high biomass, the bubbles may coalesce resulting in the reduction of
gas-liquid interface area (Huang and McDonald 2012). BCRs are liquid-phase bio-
reactors in which the roots are submerged in the medium. Liquid mixing is obtained
by the upflow of air bubbles generated from an air distributor situated at the bottom
of the column. In contrast to BCRs, airlift reactors (ALRs) contain a draft tube
(either internal or external) to avoid coalescing bubbles. ALRs distribute shear stress
more evenly, reduce shear stress, consume little energy, and promote a cylindrical
mixing of the medium (Stiles and Liu 2013).
The draft tube in ALRs prevents bubble coalescence by forcing the bubbles to
move in one direction. It also distributes shear stresses equally throughout the reac-
tor. As a result, cells are able to grow in a more stable physical environment than
those growing under high shear, a condition causing cell damage and lower produc-
tivity in STRs. It has also been shown experimentally that shear stress rates gener-
ated in ALRs are lower than those generated in BCRs.
ALRs have been extensively used for HRs since the initiation of HR bioreactor
studies for species including Panax ginseng (Yoshikawa and Furuya 1987),
Armoracia rusticana (Taya et al. 1989), Trigonella foenum-graceum (Rodriguez-
Mendiola et al. 1991), Lippia dulcis (Sauerwein et al. 1991), Lithospermum eryth-
rorhizon (Shimomura et al. 1991), Ophiorrhiza pumila (Sudo et al. 2002), and
Echinacea purpurea (Abbasi et al. 2009).
Conventional ALRs have been extensively used for scale-up cultures of HR
lines; however, they are generally not appropriate for high-density cultures due to
inadequate mixing and oxygen mass transfer (Choi et al. 2008). This phenomenon
is mainly based on uneven distribution of root tissue at certain regions as well as
excessive gas-phase channeling (Taya et al. 1989).
9 l NMRs by Huang et al. (2004) to evaluate the oxygen uptake rate, effects of inter-
mittent medium supply, and other growth-related parameters.
Several other comparative studies also have been evaluated the optimal bioreac-
tor type for a particular species of HRs. The production of Artemisia annua HRs
was compared both in bubble and mist bioreactors. Based on the results, authors
suggested that the overall biomass was higher in the BCR (Kim et al. 2002a, b).
However, the mist reactor usually accumulates lower amount of biomass as com-
pared to the BCR. This behavior could be due to insufficient nutrient availability
(Choi et al. 2008; Srivastava and Srivastava 2012a, b).
Process intensification methods could also be utilized in plant and tissue culture
works (Stiles and Liu 2013). The ability to exploit HR cultures as a source of bioac-
tive chemicals depends on the development of a suitable bioreactor system where
several physical and chemical parameters must be taken into consideration. Selection
of highly productive cell lines, manipulation of nutrients, optimizing the culture
environment, elicitation, metabolic engineering, in situ product removal, and ultra-
sound have been applied for process intensification in HR bioreactor cultures
(Mishra and Ranjan 2008; Stiles and Liu 2013).
7.6.2.2 Elicitation
Elicitation is the effective technique which is currently used for improving the pro-
duction of secondary metabolites (Zhao et al. 2005; Baenas et al. 2014). Overall,
based on origin, elicitors are classified biotic and abiotic. Basically, biotic elicitors
are either physical factors or chemical factors such as ultraviolet light heavy metals
and salts (Stiles and Liu 2013). Salts including AlCl3, AgNO3, CdCl2, CaCl2, CuCl2,
CoCl2, KCl, HgCl2, MgSO4, VOSO4NiSO4, and Zn ions have been used to increase
the secondary metabolite production in a variety of plant (Ramirez-Estrada et al.
2016; Li et al. 2006; Vasconsuelo and Boland 2007). Abiotic elicitors are usually
cheaper than biotic; however, they are not as efficient for the cultivation of the target
microorganism (Georgiev et al. 2007). Specificity of the elicitor, culture growth
stage, treatment interval, the concentration, medium composition, and light are the
main factors that affect the effectiveness of elicitation (Sharma et al. 2013).
To date, significant progresses have been made in the genetic transformation and
tissue culture in order to amplify the key pathways for the biosynthesis of targeted
metabolites. Commercial production of HRs has attracted much attention recently
as compared to the other plant cells. HR cultures are unique due to their much
higher genetic and biosynthetic stability. However, exploration into inexpensive
novel elicitors and bioreactors are required in order to warrant their industrial imple-
mentation. Further, generated knowledge from plant metabolic pathways and
advancements in genetic engineering will help HRs to become a promising and
sustainable fabrication system in the near future.
150 H. Vaghari et al.
Acknowledgments The authors would like to thank Food Research Institute of Sahand University
of Technology for cooperation in preparation of this review.
References
Abbasi BH, Tian CL, Murch SJ, Saxena PK, Liu CZ (2007) Light-enhanced caffeic acid deriva-
tives biosynthesis in hairy root cultures of Echinacea purpurea. Plant Cell Rep 26:1367–1372
Abbasi BH, Liu R, Saxena PK, Liu CZ (2009) Cichoric acid production from hairy root cultures
of Echinacea purpurea grown in a modified airlift bioreactor. J Chem Technol Biotechnol
84:1697–1701
Agostini E, Coniglio MS, Milrad SR, Tigier HA, Giulietti AM (2003) Phytoremediation of
2,4-dichlorophenol by Brassica napus hairy root cultures. Biotechnol Appl Biochem
37:139–144
Ahmad M, Nadeem SM, Naveed M, Zahir ZA (2016) Potassium-solubilizing bacteria and their
application in agriculture. In: Meena VS, Maurya BR, Verma JP, Meena RS (eds) Potassium
solubilizing microorganisms for sustainable agriculture. Springer, India, pp 293–313.
doi:10.1007/978-81-322-2776-2_21
Alderete LG, Talano MA, Ibáñez SG, Purro S, Agostini E, Milrad SR, Medina MI (2009)
Establishment of transgenic tobacco hairy roots expressing basic peroxidases and its applica-
tion for phenol removal. J Biotechnol 139:273–279
Angelini VA, Orejas J, Medina MI, Agostini E (2011) Scale up of 2, 4-dichlorophenol removal
from aqueous solutions using Brassica napus hairy roots. J Hazard Mater 185:269–274
Ayora-Talavera T, Chappell J, Lozoya-Gloria E, Loyola-Vargas VM (2002) Overexpression in
Catharanthus roseus hairy roots of a truncated hamster 3-hydroxy-3-methylglutaryl-CoA
reductase gene. Appl Biochem Biotechnol 97:135–145
Azlan GJ, Marziah M, Radzali M, Johari R (2002) Establishment of Physalis minima hairy roots
culture for the production of physalins. Plant Cell Tissue Organ Cult 69:271–278
Baenas N, García-Viguera C, Moreno DA (2014) Elicitation: a tool for enriching the bioactive
composition of foods. Molecules 19:13541–13563
Bahadur I, Meena VS, Kumar S (2014) Importance and application of potassic biofertilizer in
Indian agriculture. Int Res J Biol Sci 3:80–85
Bahadur I, Maurya BR, Kumar A, Meena VS, Raghuwanshi R (2016a) Towards the soil sustain-
ability and potassium-solubilizing microorganisms. In: Meena VS, Maurya BR, Verma JP,
Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture. Springer,
India, pp 225–266. doi:10.1007/978-81-322-2776-2_18
Bahadur I, Maurya BR, Meena VS, Saha M, Kumar A, Aeron A (2016b) Mineral release dynamics
of tricalcium phosphate and waste muscovite by mineral-solubilizing rhizobacteria isolated
from indo-gangetic plain of India. Geomicrobiol J. doi:10.1080/01490451.2016.1219431
Bahadur I, Maurya BR, Meena VS, Saha M, Kumar A, Aeron A (2017) Mineral release dynamics
of tricalcium phosphate and waste muscovite by mineral-solubilizing rhizobacteria isolated
from indo-gangetic plain of India. Geomicrobiol J. doi:10.1080/01490451.2016.1219431
Banerjee S, Shang TQ, Wilson AM, Moore AL, Strand SE, Gordon MP, Lafferty Doty S (2002)
Expression of functional mammalian P450 2E1 in hairy root cultures. Biotechnol Bioeng
77:462–466
Boominathan R, Doran PM (2003) Cadmium tolerance and antioxidative defenses in hairy roots of
the cadmium hyperaccumulator, Thlaspi caerulescens. Biotechnol Bioeng 83:158–167
Boominathan R, Saha-Chaudhury NM, Sahajwalla V, Doran PM (2004) Production of nickel bio-
ore from hyperaccumulator plant biomass: applications in phytomining. Biotechnol Bioeng
86:243–250
Bordonaro JL, Curtis WR (2000) Inhibitory role of root hairs on transport within root culture bio-
reactors. Biotechnol Bioeng 70:176–186
7 Hairy Root Culture: A Biotechnological Approach to Produce Valuable Metabolites 151
Chandra S (2012) Natural plant genetic engineer Agrobacterium rhizogenes: role of T-DNA in
plant secondary metabolism. Biotechnol Lett 34:407–415
Chandra S, Chandra R (2011) Engineering secondary metabolite production in hairy roots.
Phytochem Rev 10:371–395
Chinou I (2008) Primary and secondary metabolites and their biological activity. In: Waksmundzka-
Hajnos M, Sherma J, Kowalska T (eds) Thin layer chromatography in photochemistry. CRC
Press, Boca Raton, pp 59–76
Choi YE, Kim YS, Paek KY (2008) Types and designs of bioreactors for hairy root culture. In:
Gupta SD, Ibaraki Y (eds) Plant tissue culture engineering. Springer, Netherlands, pp 161–172
Coniglio MS, Busto VD, González PS, Medina MI, Milrad S, Agostini E (2008) Application of
Brassica napus hairy root cultures for phenol removal from aqueous solutions. Chemosphere
72:1035–1042
Curtis WR (2000) Bioreactor growth of hairy roots. In: Spier R (ed) Encyclopedia of cell technol-
ogy. Wiley, New York, pp 827–841
Das I, Pradhan M (2016) Potassium-solubilizing microorganisms and their role in enhancing
soil fertility and health. In: Meena VS, Maurya BR, Verma JP, Meena RS (eds) Potassium
solubilizing microorganisms for sustainable agriculture. Springer, India, pp 281–291.
doi:10.1007/978-81-322-2776-2_20
Datta A, Bhattacharya S, Pal CB, Sen J, Dasgupta S, Biswas A, Batra J (2010) Process for
production of anti-diabetic compound in root culture of Catharanthus roseus.US Patent
WO2010004584 Aug 2010
De Araujo BS, Dec J, Bollag JM, Pletsch M (2006) Uptake and transformation of phenol and chlo-
rophenols by hairy root cultures of Daucus carota, Ipomoea batatas and Solanum aviculare.
Chemosphere 63:642–651
Dechaux CE, Boitel-Conti MI (2005) A strategy for overaccumulation of scopolamine in Datura
innoxia hairy root cultures. Acta Biol Cracov Ser Bot 47:101–107
Dixit AK, Vaidya S (2010) Agrobacterium rhizogenes induced hairy root development and its
effect on production of glycyrrhizin in Abrus precatorius (L). Int J Curr Res 6:33–38
Dominguez-Nunez JA, Benito B, Berrocal-Lobo M, Albanesi A (2016) Mycorrhizal fungi: role
in the solubilization of potassium. In: Meena VS, Maurya BR, Verma JP, Meena RS (eds)
Potassium solubilizing microorganisms for sustainable agriculture. Springer, India, pp 77–98.
doi:10.1007/978-81-322-2776-2_6
Doran PM (2009) Application of plant tissue cultures in phytoremediation research: incentives and
limitations. Biotechnol Bioeng 103:60–76
Dotaniya ML, Meena VD, Basak BB, Meena RS (2016) Potassium uptake by crops as well
as microorganisms. In: Meena VS, Maurya BR, Verma JP, Meena RS (eds) Potassium
solubilizing microorganisms for sustainable agriculture. Springer, India, pp 267–280.
doi:10.1007/978-81-322-2776-2_19
Doty SL (2008) Enhancing phytoremediation through the use of transgenics and endophytes. New
Phytol 179:318–333
Eapen S, Suseelan KN, Tivarekar S, Kotwal SA, Mitra R (2003) Potential for rhizofiltration of ura-
nium using hairy root cultures of Brassica juncea and Chenopodium amaranticolor. Environ
Res 91:127–133
Eapen S, Singh S, D’souza SF (2007) Advances in development of transgenic plants for remedia-
tion of xenobiotic pollutants. Biotechnol Adv 25:442–451
Eibl R, Eibl D (2006) Design and use of the wave bioreactor for plant cell culture. In: Gupta SD,
Ibaraki Y (eds) Plant tissue culture engineering. Springer, The Netherlands, pp 203–227
Eibl R, Eibl D (2008) Design of bioreactors suitable for plant cell and tissue cultures. Phytochem
Rev 7:593–598
Ferdosi A, Kashefi B (2014) A review hairy roots and secondary metabolite production in salvia.
Intl J Farm Allied Sci 3:940–945
Flores HE, Filner P (2012) Metabolic relationships of putrescine, GABA and alkaloids in cell and
root cultures of Solanaceae. In: Neumann KH, Barz W, Reinhard E (eds) Primary and second-
ary metabolism of plant cell cultures. Springer Science and Business Media, p 174–185
152 H. Vaghari et al.
Fu CX, Zhao DX, Xue XF, Jin ZP, Ma FS (2005) Transformation of Saussurea involucrata by
Agrobacterium rhizogenes: hairy root induction and syringin production. Process Biochem
40:3789–3794
Garbisu C, Alkorta I (2001) Phytoextraction: a cost-effective plant-based technology for the
removal of metals from the environment. Bioresour Technol 77:229–236
Georgiev M, Heinrich M, Kerns G, Pavlov A, Bley T (2006) Production of iridoids and phenolics
by transformed Harpagophytum procumbens root cultures. Eng Life Sci 6:593–596
Georgiev MI, Pavlov AI, Bley T (2007) Hairy root type plant in vitro systems as sources of bioac-
tive substances. Appl Microbiol Biotechnol 74:1175–1185
Georgiev MI, Ludwig-Müller J, Bley T (2010) Hairy root culture: copying nature in new biopro-
cesses. In: Arora R (ed) Medicinal plant biotechnology. CAB International, Oxon, pp 156–175
Georgiev MI, Ludwig-Müller J, Alipieva K, Lippert A (2011) Sonication-assisted Agrobacterium
rhizogenes-mediated transformation of Verbascum xanthophoeniceum Griseb for bioactive
metabolite accumulation. Plant Cell Rep 30:859–866
Giri A, Narasu ML (2000) Transgenic hairy roots: recent trends and applications. Biotechnol Adv
18:1–22
Giri A, Ravindra ST, Dhingra V, Narasu ML (2001) Influence of different strains of Agrobacterium
rhizogenes on induction of hairy roots and artemisinin production in Artemisia annua. Curr
Sci 81:378–382
Gonzalez PS, Capozucca C, Tigier H, Milrad S, Agostini E (2006) Phytoremediation of phe-
nol from wastewater, by peroxidases of tomato hairy root cultures. Enzym Microb Technol
39:647–653
Gonzalez PS, Agostini E, Milrad SR (2008) Comparison of the removal of 2,4-dichlorophenol and
phenol from polluted water, by peroxidases from tomato hairy roots, and protective effect of
polyethylene glycol. Chemosphere 70:982–989
Gonzalez PS, Ontanon OM, Armendariz AL, Talano MA, Paisio CE, Agostini E (2013) Brassica
napus hairy roots and rhizobacteria for phenolic compounds removal. Environ Sci Pollut Res
20:1310–1317
Guillon S, Trémouillaux-Guiller J, Pati PK, Rideau M, Gantet P (2006) Hairy root research: recent
scenario and exciting prospects. Curr Opin Plant Biol 9:341–346
Gujarathi NP, Haney BJ, Park HJ, Wickramasinghe SR, Linden JC (2005) Hairy roots of
Helianthus annuus: a model system to study phytoremediation of tetracycline and oxytetracy-
cline. Biotechnol Prog 21:775–780
Huang TK, McDonald KA (2012) Bioreactor systems for in vitro production of foreign proteins
using plant cell cultures. Biotechnol Adv 30:398–409
Huang SY, Hung CH, Chou SN (2004) Innovative strategies for operation of mist trickling reactors
for enhanced hairy root proliferation and secondary metabolite productivity. Enzym Microb
Technol 35:22–32
Huber C, Bartha B, Harpaintner R, Schröder P (2009) Metabolism of acetaminophen (paracetamol)
in plants-two independent pathways result in the formation of a glutathione and a glucose con-
jugate. Environ Sci Pollut Res 16:206–213
Ibanez SG, Medina MI, Agostini E (2011) Phenol tolerance, changes of antioxidative enzymes and
cellular damage in transgenic tobacco hairy roots colonized by Arbuscular mycorrhizal fungi.
Chemosphere 83:700–705
Ibanez S, Talano M, Ontañon O, Suman J, Medina MI, Macek T, Agostini E (2016) Transgenic
plants and hairy roots: exploiting the potential of plant species to remediate contaminants. New
Biotechnol 33:625–635
Ismail AM, Theodor PA (2012) The effect of heavy metals Zn and Ni on growth of in vitro hairy
root cultures of Indian mustard Brassica juncea L. Int J Adv Biotechnol Res 3:688–697
Jacob A, Malpathak N (2004) Green hairy root cultures of Solanum khasianum Clarke-a new route
to in vitro Solasodine production. Curr Sci 87:1442–1447
Jaiswal DK, Verma JP, Prakash S, Meena VS, Meena RS (2016) Potassium as an important plant
nutrient in sustainable agriculture: a state of the art. In: Meena VS, Maurya BR, Verma JP,
7 Hairy Root Culture: A Biotechnological Approach to Produce Valuable Metabolites 153
Kumar A, Meena VS, Maurya BR, Raghuwanshi R, Bisht JK, Pattanayak A (2017b) Towards the
biological nitrogen fixation and nitrogen management in legume under sustainable agriculture.
Appl Soil Ecol. doi:10.1016/j.apsoil.2017.05.013
Kwok KH, Doran PM (1995) Kinetic and stoichiometric analysis of hairy roots in a segmented
bubble-column reactor. Biotechnol Prog 11:429–435
Lan X, Quan H (2010) Hairy root culture of Przewalskia tangutica for enhanced production of
pharmaceutical tropane alkaloids. J Med Plant Res 4:1477–1481
Le Flem-Bonhomme V, Laurain-Mattar D, Fliniaux MA (2004) Hairy root induction of Papaver
somniferum var. album, a difficult-to-transform plant, by A. rhizogenes LBA 9402. Planta
218:890–893
Li W, Koike K, Asada Y, Hirotani M, Rui H, Yoshikawa T, Nikaido T (2002) Flavonoids from
Glycyrrhiza pallidiflora hairy root cultures. Phytochemistry 60:351–355
Li FX, Jin ZP, Zhao DX, Cheng LQ, Fu CX, Ma FS (2006) Overexpression of the Saussurea
medusa chalcone isomerase gene in Saussurea involucrata hairy root cultures enhances their
biosynthesis of apigenin. Phytochemistry 67:553–560
Lokhande VH, Kudale S, Nikalje G, Desai N, Suprasanna P (2015) Hairy root induction and phy-
toremediation of textile dye, reactive green 19A-HE4BD, in a halo-phyte, Sesuvium portulac-
astrum L. Biotechnol Rep 8:56–63
Ludwig-Müller J, Jahn L, Lippert A, Püschel J, Walter A (2014) Improvement of hairy root cul-
tures and plants by changing biosynthetic pathways leading to pharmaceutical metabolites:
strategies and applications. Biotechnol Adv 32:1168–1179
Macek T, Kotrba P, Svatos A, Novakova M, Demnerova K, Mackova M (2008) Novel roles for
genetically modified plants in environmental protection. Trends Biotechnol 26:146–152
Masood S, Bano A (2016) Mechanism of potassium solubilization in the agricultural soils by
the help of soil microorganisms. In: Meena VS, Maurya BR, Verma JP, Meena RS (eds)
Potassium solubilizing microorganisms for sustainable agriculture. Springer, India, pp 137–
147. doi:10.1007/978-81-322-2776-2_10
Maurya BR, Meena VS, Meena OP (2014) Influence of Inceptisol and Alfisol’s potassium solubi-
lizing bacteria (KSB) isolates on release of K from waste mica. Vegetos 27:181–187
McCoy E, O’Connor SE (2008) Natural products from plant cell cultures. In: Petersen F, Amstutz
R (eds) Natural compounds as drugs, vol 1. Birkhäuser, Basel, pp 329–370
McKelvey SA, Gehrig JA, Hollar KA, Curtis WR (1993) Growth of plant root cultures in liquid-
and gas-dispersed reactor environments. Biotechnol Prog 9:317–322
Medina-Bolivar LF, Dolan M, Bennett S, Condori JM, Hubstenberger JF, inventors; Condori Jose
M, Hubstenberger John F, assignee (2010) Production of stilbenes in plant hairy root cultures.
US patent 7,666,677, 23 Feb 2010
Meena VS, Maurya BR, Bohra JS, Verma R, Meena MD (2013a) Effect of concentrate manure
and nutrient levels on enzymatic activities and microbial population under submerged rice in
alluvium soil of Varanasi. Crop Res 45(1, 2 & 3):6–12
Meena OP, Maurya BR, Meena VS (2013b) Influence of K-solubilizing bacteria on release of
potassium from waste mica. Agric Sust Dev 1:53–56
Meena VS, Maurya BR, Verma R, Meena RS, Jatav GK, Meena SK, Meena SK (2013c) Soil
microbial population and selected enzyme activities as influenced by concentrate manure and
inorganic fertilizer in alluvium soil of Varanasi. Bioscan 8(3):931–935
Meena VS, Maurya BR, Bahadur I (2014a) Potassium solubilization by bacterial strain in waste
mica. Bang J Bot 43:235–237
Meena VS, Maurya BR, Verma JP (2014b) Does a rhizospheric microorganism enhance K+ avail-
ability in agricultural soils? Microbiol Res 169:337–347
Meena VS, Maurya BR, Meena RS (2015a) Residual impact of wellgrow formulation and NPK
on growth and yield of wheat (Triticum aestivum L.). Bangladesh. Aust J Bot 44(1):143–146
Meena VS, Maurya BR, Verma JP, Aeron A, Kumar A, Kim K, Bajpai VK (2015b) Potassium
solubilizing rhizobacteria (KSR): isolation, identification, and K-release dynamics from waste
mica. Ecol Eng 81:340–347
7 Hairy Root Culture: A Biotechnological Approach to Produce Valuable Metabolites 155
Meena RS, Meena VS, Meena SK, Verma JP (2015c) The needs of healthy soils for a healthy
world. J Clean Prod 102:560–561
Meena RS, Meena VS, Meena SK, Verma JP (2015d) Towards the plant stress mitigate the agricul-
tural productivity: a book review. J Clean Prod 102:552–553
Meena VS, Meena SK, Verma JP, Meena RS, Ghosh BN (2015e) The needs of nutrient use efficiency
for sustainable agriculture. J Clean Prod 102:562–563. doi:10.1016/j.jclepro.2015.04.044
Meena VS, Verma JP, Meena SK (2015f) Towards the current scenario of nutrient use efficiency in
crop species. J Clean Prod 102:556–557. doi:10.1016/j.jclepro.2015.04.030
Meena VS, Bahadur I, Maurya BR, Kumar A, Meena RK, Meena SK, Verma JP (2016a) Potassium-
solubilizing microorganism in evergreen agriculture: an overview. In: Meena VS, Maurya BR,
Verma JP, Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture.
Springer, India, pp 1–20. doi:10.1007/978-81-322-2776-2_1
Meena RS, Bohra JS, Singh SP, Meena VS, Verma JP, Verma SK, Sihag SK (2016b) Towards the
prime response of manure to enhance nutrient use efficiency and soil sustainability a current
need: a book review. J Clean Prod 112(1):1258–1260
Meena VS, Meena SK, Bisht JK, Pattanayak A (2016c) Conservation agricultural practices in
sustainable food production. J Clean Prod 137:690–691
Meena SK, Rakshit A, Meena VS (2016d) Effect of seed bio-priming and N doses under varied
soil type on nitrogen use efficiency (NUE) of wheat (Triticum aestivum L.) under greenhouse
conditions. Biocatal Agric Biotechnol 6:68–75
Meena RK, Singh RK, Singh NP, Meena SK, Meena VS (2016e) Isolation of low temperature sur-
viving plant growth-promoting rhizobacteria (PGPR) from pea (Pisum sativum L.) and docu-
mentation of their plant growth promoting traits. Biocatalysis Agric Biotechnol 4:806–811
Mishra BN, Ranjan R (2008) Growth of hairy-root cultures in various bioreactors for the produc-
tion of secondary metabolites. Biotechnol Appl Biochem 49:1–10
Moyano E, Jouhikainen K, Tammela P, Palazón J, Cusidó RM, Piñol MT, Teeri TH, Oksman-
Caldentey KM (2003) Effect of pmt gene overexpression on tropane alkaloid production in
transformed root cultures of Datura metel and Hyoscyamus muticus. J Exp Bot 54:203–211
Najmanova J, Mackova M, Macek T, Kotrba P (2007) Preparation of transgenic flax with enhanced
metal tolerance. J Biotechnol 131:S38–S39
Namdeo AG (2007) Plant cell elicitation for production of secondary metabolites: a review.
Pharmacogn Rev 1:69–79
Nath D, Maurya BR, Meena VS (2017) Documentation of five potassium- and phosphorus-
solubilizing bacteria for their K and P-solubilization ability from various minerals. Biocatal
Agric Biotechnol 10:174–181
Nedelkoska TV, Doran PM (2000) Characteristics of heavy metal uptake by plant species with
potential for phytoremediation and phytomining. Miner Eng 13:549–561
Nepovim A, Podlipná R, Soudek P, Schröder P, Vanek T (2004) Effects of heavy metals and nitroaro-
matic compounds on horseradish glutathione S-transferase and peroxidase. Chemosphere
57:1007–1015
Nguyen C, Bourgaud F, Forlot P, Guckert A (1992) Establishment of hairy root cultures of Psoralea
species. Plant Cell Rep 11:424–427
Novakova M, Mackova M, Sylvestre M, Macek T (2007) Preparation of genetically modified
plants containing bacterial dioxygenase – tool for preferable phytoremediation. J Biotechnol
131:S36
Oller AL, Agostini E, Talano MA, Capozucca C, Milrad SR, Tigier HA, Medina MI (2005)
Overexpression of a basic peroxidase in transgenic tomato (Lycopersicon esculentum Mill. cv.
Pera) hairy roots increases phytoremediation of phenol. Plant Sci 169:1102–1111
Ono NN, Tian L (2011) The multiplicity of hairy root cultures: prolific possibilities. Plant Sci
180:439–446
Ontañon OM, González PS, Ambrosio LF, Paisio CE, Agostini E (2014) Rhizoremediation of
phenol and chromium by the synergistic combination of a native bacterial strain and Brassica
napus hairy roots. Int Biodeterior Biodegrad 88:192–198
156 H. Vaghari et al.
Palazón J, Cusidó RM, Bonfill M, Mallol A, Moyano E, Morales C, Piñol MT (2003a) Elicitation
of different Panax ginseng transformed root phenotypes for an improved ginsenoside produc-
tion. Plant Physiol Biochem 41:1019–1025
Palazón J, Mallol A, Eibl R, Lettenbauer C, Cusidó RM, Piñol MT (2003b) Growth and ginsen-
oside production in hairy root cultures of Panax ginseng using a novel bioreactor. Planta Med
69:344–349
Parewa HP, Yadav J, Rakshit A, Meena VS, Karthikeyan N (2014) Plant growth promoting rhizo-
bacteria enhance growth and nutrient uptake of crops. Agric Sustain Dev 2(2):101–116
Park SU, Facchini PJ (2000) Agrobacterium rhizogenes-mediated transformation of opium poppy,
Papaver somniferum L., and California poppy, Eschscholzia californica Cham, root cultures.
J Exp Bot 51:1005–1016
Patil P, Desai N, Govindwar S, Jadhav JP, Bapat V (2009) Degradation analysis of reactive Red198
by hairy roots of Tagetes patula L. (Marigold). Planta 230:725–735
Pavlov A, Georgiev V, Kovatcheva P (2003) Relationship between type and age of the inocu-
lum cultures and betalains biosynthesis by Beta vulgaris hairy root culture. Biotechnol Lett
25:307–309
Pavlov A, Berkov S, Weber J, Bley T (2009) Hyoscyamine biosynthesis in Datura stramonium
hairy root in vitro systems with different ploidy levels. Appl Biochem Biotechnol 157:210–225
Pistelli L, Giovannini A, Ruffoni B, Bertoli A, Pistelli L (2010) Hairy root cultures for secondary
metabolites production. In: Teresa M, Giuseppina G, Berra R, Berra B (eds) Bio-farms for
nutraceuticals. Springer, US, pp 167–184
Prakash S, Verma JP (2016) Global perspective of potash for fertilizer production. In: Meena VS,
Maurya BR, Verma JP, Meena RS (eds) Potassium solubilizing microorganisms for sustainable
agriculture. Springer, India, pp 327–331. doi:10.1007/978-81-322-2776-2_23
Praveen N, Murthy HN (2012) Synthesis of with anolide A depends on carbon source and medium
pH in hairy root cultures of Withania somnifera. Ind Crop Prod 35:241–243
Priyadharsini P, Muthukumar T (2016) Interactions between arbuscular mycorrhizal fungi and
potassium-solubilizing microorganisms on agricultural productivity. In: Meena VS, Maurya
BR, Verma JP, Meena RS (eds) Potassium solubilizing microorganisms for sustainable agricul-
ture. Springer, India, pp 111–125. doi:10.1007/978-81-322-2776-2_8
Qaderi A, Akbari Z, Kalateh-jari S, Fatehi F, Tolyat M, Jalali Moghadam M, Naghdi Badi H (2016)
Improving trigonelline production in hairy root culture of fenugreek (Trigonella foenum-
graecum). J Med Plants 59:73–80
Raghavendra MP, Nayaka NC, Nuthan BR (2016) Role of rhizosphere microflora in potas-
sium solubilization. In: Meena VS, Maurya BR, Verma JP, Meena RS (eds) Potassium
solubilizing microorganisms for sustainable agriculture. Springer, India, pp 43–59.
doi:10.1007/978-81-322-2776-2_4
Ramakrishnan D, Curtis WR (2004) Trickle-bed root culture bioreactor design and scale-up:
growth, fluid-dynamics, and oxygen mass transfer. Biotechnol Bioeng 88:248–260
Ramirez-Estrada K, Vidal-Limon H, Hidalgo D, Moyano E, Golenioswki M, Cusidó RM, Palazon
J (2016) Elicitation, an effective strategy for the biotechnological production of bioactive high-
added value compounds in plant cell factories. Molecules 21:182–205
Rao SR, Ravishankar GA (2002) Plant cell cultures: chemical factories of secondary metabolites.
Biotechnol Adv 20:101–153
Rates SM (2001) Plants as source of drugs. Toxicon 39:603–613
Rawat J, Sanwal P, Saxena J (2016) Potassium and its role in sustainable agriculture. In: Meena
VS, Maurya BR, Verma JP, Meena RS (eds) Potassium solubilizing microorganisms for sus-
tainable agriculture. Springer, India, pp 235–253. doi:10.1007/978-81-322-2776-2_17
Rezek J, Macek T, Mackova M, Triska J (2007) Plant metabolites of polychlorinated biphenyls in
hairy root culture of black nightshade Solanum nigrum SNC-90. Chemosphere 69:1221–1227
Rezek J, Macek T, Doubsky J, Mackova M (2012) Metabolites of 2,2′-dichlorobiphenyl and
2,6-dichlorobiphenyl in hairy root culture of black nightshade Solanum nigrum SNC-90.
Chemosphere 89:383–838
7 Hairy Root Culture: A Biotechnological Approach to Produce Valuable Metabolites 157
Richter U, Rothe G, Fabian AK, Rahfeld B, Dräger B (2005) Overexpression of tropinone reduc-
tases alters alkaloid composition in Atropa belladonna root cultures. J Exp Bot 56:645–652
Rodriguez-Mendiola MA, Stafford A, Cresswell R, Ariascastro C (1991) Bioreactors for growth of
plant-roots. Enzym Microb Technol 13:697–702
Rudrappa T, Neelwarne B, Aswathanarayana RG (2004) In situ and ex situ adsorption and recov-
ery of betalains from hairy root cultures of Beta vulgaris. Biotechnol Prog 20:777–785
Saha M, Maurya BR, Bahadur I, Kumar A, Meena VS (2016a) Can potassium-solubilising bac-
teria mitigate the potassium problems in India? In: Meena VS, Maurya BR, Verma JP, Meena
RS (eds) Potassium solubilizing microorganisms for sustainable agriculture. Springer, India,
pp 127–136. doi:10.1007/978-81-322-2776-2_9
Saha M, Maurya BR, Meena VS, Bahadur I, Kumar A (2016b) Identification and characterization
of potassium solubilizing bacteria (KSB) from Indo-Gangetic Plains of India. Biocatal Agric
Biotechnol 7:202–209
Sarkar D, Meena VS, Haldar A, Rakshit R (2017) Site-specific nutrient management (SSNM):
a unique approach towards maintaining soil health. In: The adaptive soil management: from
theory to practices. p 69–88 doi. 10.1007/978-981-10-3638-5_3
Sato F, Hashimoto T, Hachiya A, Tamura KI, Choi KB, Morishige T, Fujimoto H, Yamada Y
(2001) Metabolic engineering of plant alkaloid biosynthesis. Proc Natl Acad Sci 98:367–372
Sauerwein M, Yamazaki T, Shimomura K (1991) Hernandulcin in hairy root cultures of Lippia
dulcis. Plant Cell Rep 9:579–581
Sevón N, Oksman-Caldentey KM (2002) Agrobacterium rhizogenes-mediated transformation:
root cultures as a source of alkaloids. Planta Med 68:859–868
Sharma S, Shahzad A, Sahai A (2013) Hairy root culture: an efficient system for secondary metab-
olite production. In: Shahid M, Shahzad A, Malik A, Sahai A (eds) Recent trends in biotech-
nology and therapeutic applications of medicinal plants. Springer, The Netherlands, pp 51–78
Sharma A, Shankhdhar D, Shankhdhar SC (2016) Potassium-solubilizing microorganisms:
mechanism and their role in potassium solubilization and uptake. In: Meena VS, Maurya BR,
Verma JP, Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture.
Springer, India, pp 203–219. doi:10.1007/978-81-322-2776-2_15
Shimomura K, Sudo H, Saga H, Kamada H (1991) Shikonin production and secretion by hairy root
cultures of Lithospermum erythrorhizon. Plant Cell Rep 10:282–285
Shrivastava M, Srivastava PC, D’Souza SF (2016) KSM soil diversity and mineral solubilization,
in relation to crop production and molecular mechanism. In: Meena VS, Maurya BR, Verma JP,
Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture. Springer,
India, pp 221–234. doi:10.1007/978-81-322-2776-2_16
Sindhu SS, Parmar P, Phour M, Sehrawat A (2016) Potassium-solubilizing microorganisms
(KSMs) and its effect on plant growth improvement. In: Meena VS, Maurya BR, Verma JP,
Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture. Springer,
India, pp 171–185. doi:10.1007/978-81-322-2776-2_13
Singh S, Melo JS, Eapen S, D’souza SF (2006) Phenol removal using Brassica juncea hairy roots:
role of inherent peroxidase and H2O2. J Biotechnol 123:43–49
Singh NP, Singh RK, Meena VS, Meena RK (2015) Can we use maize (Zea mays) rhizobacteria as
plant growth promoter? Vegetos 28(1):86–99. doi:10.5958/2229-4473.2015.00012.9
Singh M, Dotaniya ML, Mishra A, Dotaniya CK, Regar KL, Lata M (2016) Role of biofertil-
izers in conservation agriculture. In: Bisht JK, Meena VS, Mishra PK, Pattanayak A (eds)
Conservation agriculture: an approach to combat climate change in Indian Himalaya. Springer,
Singapore, pp 113–134. doi:10.1007/978-981-10-2558-7_4
Sivakumar G, Yu KW, Hahn EJ, Paek KY (2005) Optimization of organic nutrients for ginseng
hairy roots production in large-scale bioreactors. Curr Sci 89:641–649
Sivanesan I, Jeong BR (2009) Induction and establishment of adventitious and hairy root cultures
of Plumbago zeylanica L. Afr J Biotechnol 8:5294–5300
Sosa Alderete LG, Talano MA, Ibáñez SG, Purro S, Agostini E, Milrad SR et al (2009) Establishment
of transgenic tobacco hairy roots expressing basic peroxidases and its application for phenol
removal. J Biotechnol 139:273–279
158 H. Vaghari et al.
Sosa Alderete LG, Ibǎne˜z SG, Agostini E, Medina MI (2012) Phytoremediation of phenol at pilot
scale by tobacco hairy roots. Int J Environ Sci 3:398–407
Soudek P, Petrová S, Benesova D, Vanek T (2011) Uranium uptake and stress responses of in vitro
cultivated hairy root culture of Armoracia rusticana. Agrochimica 55:15–28
Srivastava S, Srivastava AK (2007) Hairy root culture for mass-production of high-value second-
ary metabolites. Crit Rev Biotechnol 27:29–43
Srivastava S, Srivastava AK (2012a) Azadirachtin production by hairy root cultivation of
Azadirachta indica in a modified stirred tank reactor. Bioprocess Biosyst Eng 35:1549–1553
Srivastava S, Srivastava AK (2012b) In vitro Azadirachtin production by hairy root cultivation of
Azadirachta indica in nutrient mist bioreactor. Appl Biochem Biotechnol 166:365–378
Srivastava S, Srivastava AK (2012c) Statistical medium optimization for enhanced azadirachtin
production in hairy root culture of Azadirachta indica. In Vitro Cell Dev Biol Plant 48:73–84
Stiles AR, Liu CZ (2013) Hairy root culture: bioreactor design and process intensification. In:
Doran PM (ed) Biotechnology of hairy root systems. Springer, Berlin, pp 91–114
Straczek A, Wannijn J, Van Hees M, Thijs H, Thiry Y (2009) Tolerance of hairy roots of carrots to
U chronic exposure in a standardized in vitro device. Environ Exp Bot 65:82–89
Sudha CG, Reddy BO, Ravishankar GA, Seeni S (2003) Production of ajmalicine and ajmaline
in hairy root cultures of Rauvolfia micrantha Hook f., a rare and endemic medicinal plant.
Biotechnol Lett 25:631–636
Sudo H, Yamakawa T, Yamazaki M, Aimi N, Saito K (2002) Bioreactor production of camptoth-
ecin by hairy root cultures of Ophiorrhiza pumila. Biotechnol Lett 24:359–363
Sung LS, Huang SY (2006) Lateral root bridging as a strategy to enhance L-DOPA production in
Stizolobium hassjoo hairy root cultures by using a mesh hindrance mist trickling bioreactor.
Biotechnol Bioeng 94:441–447
Suresh B, Sherkhane PD, Kale S, Eapen S, Ravishankar GA (2005) Uptake and degradation of DDT
by hairy root cultures of Cichorium intybus and Brassica juncea. Chemosphere 61:1288–1292
Talano MA, Frontera S, González P, Medina MI, Agostini E (2010) Removal of 2,4- diclorophenol
from aqueous solutions using tobacco hairy root cultures. J Hazard Mater 176:784–791
Talano MA, Oller AL, Gonzalez PS, Agostini E (2012) Hairy roots, their multiple applications and
recent patents. Recent Patents Biotechnol 6:115–133
Talano MA, Oller AL, González P, González SO, Agostini E (2014) Effects of arsenate on tobacco
hairy root and seedling growth, and its removal. In Vitro Cell Dev Biol Plant 50:217–225
Taya M, Yoyama A, Kondo O, Kobayashi T, Matsui C (1989) Growth characteristics of plant hairy
roots and their cultures in bioreactors. J Chem Eng Jpn 22:84–89
Taya M, Sato H, Kinooka M, Tone S (1994) Characterization of Pak-bung green hairy roots culti-
vated under light irradiation. J Ferment Bioeng 78:42–48
Telke AA, Kagalkar AN, Jagtap UB, Desai NS, Bapat VA, Govindwar SP (2011) Bio-chemical
characterization of laccase from hairy root culture of Brassica juncea L. and role of redox
mediators to enhance its potential for the decolorization of textile dyes. Planta 234:1137–1149
Teotia P, Kumar V, Kumar M, Shrivastava N, Varma A (2016) Rhizosphere microbes: potassium
solubilization and crop productivity-present and future aspects. In: Meena VS, Maurya BR,
Verma JP, Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture.
Springer, India, pp 315–325. doi:10.1007/978-81-322-2776-2_22
Thwe A, Arasu MV, Li X, Park CH, Kim SJ, Al-Dhabi NA, Park SU (2016) Effect of different
agrobacterium rhizogenes strains on hairy root induction and phenylpropanoid biosynthesis in
tartary buckwheat (Fagopyrum tataricum gaertn). Front Microbiol 7:318–328
Ur Rahman L, Ikenaga T, Kitamura Y (2004) Penicillin derivatives induce chemical structure-
dependent root development, and application for plant transformation. Plant Cell Rep
22:668–677
Vanisree M, Lee CY, Lo SF, Nalawade SM, Lin CY, Tsay HS (2004) Studies on the production of
some important secondary metabolites from medicinal plants by plant tissue cultures. Bot Bull
Acad Sin 45:1–22
Vasconsuelo A, Boland R (2007) Molecular aspects of the early stages of elicitation of secondary
metabolites in plants. Plant Sci 172:861–875
7 Hairy Root Culture: A Biotechnological Approach to Produce Valuable Metabolites 159
P.V. Bramhachari, Ganji Purnachandra Nagaraju,
and E. Kariali
Abstract
The rationale of this article especially emphasizes the potential contributions of
metagenomics to the study of plant growth-promoting rhizobacteria (PGPRs)
which play an important role in the sustainable agriculture. Several studies are
currently focused to unravel the molecular mechanism implicated in plant–
microbe interactions to comprehend the functionality of PGPRs. In this context,
researchers are presently using advanced molecular techniques, modern sequenc-
ing technologies, and metagenomics, to explore the structural and functional
aspects of genes that analyze the protein composition. Metagenomics has a huge
prospective to endow with fundamental knowledge on plant–microbe interac-
tions necessary for new innovations to increase sustainable agricultural produc-
tivity. This review summarizes the main features of PGPRs in sustainable
agriculture and gives a brief outlook on the recent trends in plant–microbe
metagenomics. It also critically discusses the current knowledge of plant–soil–
microbe-mediated interactions and the impact of enhanced genomic technolo-
gies and our perception to understand how these relationships impact plant
performance and sustainable improvement of plant productivity. Metagenomic
applications in framework of plant–microbe interactions are also highlighted that
profit from these novel technologies.
Keywords
Plant growth-promoting rhizobacteria (PGPR) • Sustainable agriculture • Crop
productivity • Plant–microbe interactions • Metagenomics
8.1 Introduction
The rhizosphere is a biologically dynamic zone of the soil contiguous to plant roots
that encompass soilborne microbes, where the biochemical and physical character-
istics together influence the roots (Zeyaullah et al. 2009). Studies on the plant–
microbe interactions in the rhizosphere are greatly imperative for understanding an
array of intrinsic metabolic and physiological processes, such as nutrient geocy-
cling, carbon sequestration, and ecosystem functioning. There are aplethora of
reports on the role of rhizosphere–microbe interactions in nutrient cycling; plant
diversity and carbon sinks raised noteworthy interest and enthused further researches
on plant–microbe interactions (Opik et al. 2003; Bahadur et al. 2017; Verma et al.
2017b; Kumar et al. 2017a). The microbial diversity present in soil is exceptionally
incredible, with ~109 bacteria per gram of soil representing surplus 5000 bacterial
species (Fierer et al. 2012). However, soil microbiome is very essential in shaping
the following parameters, viz., plant nutrition plan, health/disease, soil gene pool,
pesticide/pollutant decomposition, bioindicators, nutrient cycling, soil fertility, and
soil structure. Each gram soil contains 109–10 bacterial cells (107–8culturable, 104
species).
The soil metagenome has a greater biodiversity than any other habitat,
>1,000,000 Gb per gram soil estimated from 109 bacterial cells with 5 Mb genomes),
when compared to 6 Gb Sargasso sea metagenome sequenced, 3 Gb human genome,
gut metagenome 0.5 Gb, 577 Gb sequenced from 124 individual gut microbiomes
(Qin et al. 2010). Nonetheless, the ecologists face a colossal challenge to link
microbial diversity studies in the rhizosphere. Primarily the molecular interactions
between microbes and plants have been contemplated as an effect. However, as
indicated in the recent literature, it was recognized as an effective process with high
level of complexity wherein at least two genomes share information devoid of shar-
ing the same ecological niches from a cellular perspective (Sharma et al. 2016a, b).
More recently, the extent of influence among the organisms involved in the interac-
tions is therefore claimed important that the biotechnological management of
microorganisms has evicted to be an indispensable target for improving the func-
tions of plant (Swarupa et al. 2016). Therefore, the technologies that overcome
these limitations in agricultural production are highly inevitable. Several solutions
that can increase agricultural yield include better agricultural practices and land
management, use of biofertilizers, integrated pest management and farm mechani-
zation, transgenic crops, and extended use of plant growth-promoting rhizobacteria
(Glick 2014). Numerous approaches will only have short stint benefits as the world
is finite with limited resources and, they count to several environmental reparations.
Sustainable agriculture devoid of causing environmental damage is one of the key
8 Metagenomic Approaches in Understanding the Mechanism and Function… 165
Approximately 15% of the root surface is covered by microbes that belong to differ-
ent microbial species (Haldar and Sengupta 2015). The roots secrete a variety of
different sugars produced by photosynthesis (~5 to 30%) which are sequentially
utilized by rhizosphere microbes (Glick 2014). Bacteria that efficiently colonize the
rhizosphere and stimulate plant growth through direct or indirect mechanisms are
referred to as PGPRs (Ahemad and Kibretm 2014). PGPRs include a group of natu-
rally occurring soil microbial flora that inhabit in the rhizosphere and thus inflict
beneficial effect on the overall growth of the plant. PGPRs include bacteria that
belong to genera Agrobacterium, Arthrobacter, Acinetobacter, Azospirillum,
Azotobacter, Bradyrhizobium, Burkholderia, Frankia, Serratia, Rhizobium,
Pseudomonads, Thiobacillus and Bacillus (Vessey 2003). These bacteria assist the
plants by employing nutrient transport of minerals and uptake through roots (Glick
1995). Several researchers are actively involved in understanding the underlying
mechanism of plant growth promotion by PGPRs (Fig. 8.1). These mechanics are
broadly divided into direct and indirect mechanisms. Nonetheless the direct mecha-
nism involves the straight use of required nutrients and/or growth regulators for
promoting plant growth. However, if the plants were protected from various stresses
factors (biotic and abiotic stress), their healthy growth may come under indirect
mechanics (Prakash and Verma 2016; Bahadur et al. 2016; Masood and Bano 2016;
Meena et al. 2015e, 2016e; Teotia et al. 2016).
The direct mechanism of PGPRs includes indole-3-acetic acid (IAA), sidero-
phores (Jahanian et al. 2012), 1-amino-cyclopropane-1-carboxylate (ACC) deami-
nase, hydrogen cyanate (HCN), nitrogenase (Glick 2012), and phosphate
solubilization (Ahemad and Khan 2012). Perhaps some PGPRs also possess more
environment-specific PGP traits such as heavy metal-detoxifying activity (Ma et al.
2011), salinity tolerance (Tank and Saraf 2010), and biocontrol of phytopathogens
and insects (Hynes et al. 2008). PGPRs have become of interest as inoculants for
phytoremediation because of their diverse plant growth-promoting capabilities (Liu
et al. 2013). Among these, phytohormone production by PGPRs plays a very
8 Metagenomic Approaches in Understanding the Mechanism and Function… 167
PGPRs Bacteria
and fungi Plant
health/disease
Po4 Solubilzing
microbes, phytohormones,
Bioindicators
siderophores
Other endophytic
Soil fertility/Soil
bacteria, heterotrophic SUSTAINABLE structure
bacteria AGRICULTURE
important role in promoting plant growth as phytohormones can intercede with the
processes including cell division, plant cell enlargement, and extension in symbiotic
as well as nonsymbiotic roots (Glick 2014). Systematic analysis of whole genome
data and the identification of genes that contribute to the beneficial activity of PGPR
will aid our understanding of the molecular mechanisms of many bacterial species
and also help in the development of PGPR-assisted phytoremediation technology
(Fig. 8.1). NGS have recently been employed to study the genomes of several
PGPRs such as Pseudomonas sp. (Duan et al. 2013), Bacillus sp. (Song et al. 2012),
and P. polymyxa (Li et al. 2014). A thorough understanding of the plant growth-
promoting (PGP) metagenomics is highly inevitable to explore the rhizospheric
flora in order to maximize the processes that robustly augment plant productivity.
In the analysis of whole microbiome, the initial effort was started with the discovery
of a conserved 16S rRNA gene sequence and its PCR applications in the identifica-
tion of microorganisms (Woese and Fox 1977). Thus far, there are meticulous
improvements achieved with these techniques, consenting to metagenomics, in
order to study and understand the microbiome in a holistic insight in a petite period.
The comprehensive assessment of soil ecosystems and more significantly functional
genes allied with ecosystem processes are fastly curved into cost-efficient facets.
This is primarily through the development of a range of whole metagenome sam-
pling, followed by purification, separation, sequencing, and finally data analysis and
interpretation. Particularly, the sequencing technology is going through speedy
development, as it provides wide and thorough views of metagenomics, and
170 P.V. Bramhachari et al.
Soil-plant-microbes
Rhizosphere
Soil Enrichment Isolations;
Nucleic acid extractions
Function driven Cloning of DNA fragments Sequence driven
Metagenomics Metagenomics
Soil Metagenomic library
constructions
Screening of Recombinants
Sustainable
Agriculture
Fig. 8.2 Scheme of metagenomics for the discovery of novel soil biocatalysts and gene clusters
from soil microorganisms
nowadays the largely named are (a) high-throughput sequencing (HTS) and (b)
next-generation sequencing (NGS) technologies. These HTS techniques include the
use of 454 Genome Sequencer (Roche Diagnostics), the HiSeq 2000 (Illumina), and
the AB SOLiD System (Life Technologies Rincon-Florez et al. 2013; Yergeau et al.
2014; Saha et al. 2016a; Sharma et al. 2016; Verma et al. 2015a; Meena et al. 2013b).
Furthermore, other advanced techniques, such as DNA/RNA-SIP and DNA arrays
(PhyloChip and functional gene arrays), moreover have promising features in the
analysis of microbiomes, particularly their functional parts (Rincon-Florez et al.
2013; Uhlik et al. 2013). At present, there is a transition from metagenomics to
metatranscriptomics, as the latter answers the diversity and functional part of the
microbiome, rather than only showing the diversity. Nonetheless, it was also appar-
ent in recent times that the functional flexibility and function-based diversity of the
microbiome may possibly be central factors in niches somewhat than mere diversity
(Barret et al. 2011; Chaparro et al. 2012).
However, in metatranscriptomics approaches, RNA-SIP, quantitative reverse
transcription-PCR, and complementary DNA analysis coupled with pyrosequenc-
ing provide advanced functional insights into microbiome activities in the soil and
rhizosphere (Schauss et al. 2009; Uhlik et al. 2013). Predominantly, the significance
of RNA-SIP was underlined in forthcoming studies for sequential analysis of the
flow of root-derived carbon and differentiation of primary and secondary microbial
utilizers, which have elevated rates of labeling than their genes and need not depend
on cell division, unlike DNA-SIP (Bressan et al. 2009; Uhlik et al. 2013). In order
8 Metagenomic Approaches in Understanding the Mechanism and Function… 171
phosphate solubilization activity. Interestingly the genes and operons with homol-
ogy to phosphorus uptake, regulatory system and solubilization mechanisms were
investigated.
The phyllosphere metagenomic datasets generated in these studies were addi-
tionally characterized in combination with a metagenomic dataset from Tamarix-
associated phyllosphere bacteria to screen for photosynthetic genes that were further
identified from other microorganisms to be involved in light-driven energy genera-
tion and diverse microbial rhodopsins (Atamna-Ismaeel et al. 2012a, b). Another
metagenomic project was performed with the aim to attain a full sequence of a
complete genome sequence of uncultured phytopathogen and insect symbiont
Candidatus Liberibacter asiaticus which causes citrus huanglongbing disease
(Duan et al. 2009).
In the rhizosphere makes a source of gene pool with a huge potential, particularly
for agricultural applications with the aim to improve crop productivity and quality
of agricultural products and shield crops from pests. Perhaps this could be per-
formed by exploring the genes responsible for PGPR activities and application of
the recombinant molecules to soil. A functionally competent chitinase may perhaps
be used as an effective fungicide in a research experiment by Ikeda et al. (2007),
who used T-RFLP and clone library analysis to identify chitinase genes in maize
rhizosphere. The same group of researchers established novel bacterial chitinase
genes and found significantly large differences in chitinase gene diversity between
the bulk and rhizosphere soils.
An additional example of Hjort et al. (2010) also explored the potential of chitin-
ases in phytopathogen-suppressive soils. An activity that improves the plant fitness
and, hence, improves crop production is apparently ACC deaminase activity.
Interestingly Nikolic et al. (2011) also analyzed ACC deaminase genes (acdS) of
bacterial endophytes colonizing field-grown potato plants and discovered the pres-
ence of two unique types of acdS genes, the dominant one showing high homology
to an acdS gene derived from P. fluorescens through PCR analysis. The fundamental
study on siderophores is mainly fascinating due to its triple function application,
nutritional, systemic resistance inductor (ISR) and biocontrol (Bakker et al. 2007;
Ramos Solano et al. 2008). Despite its significance in agriculture, the only instance
to date that has been reported in marine settings, through expression-dependent
methods, and resulted in a cluster composed by five ORFs that synthesized vibrio-
ferrin, a siderophore explored in V. parahaemolyticus (Fujita et al. 2011).
The sequencing of whole genomes from a number of species permits to delineate
their organization and provides the basis for understanding their functionality
(Morrell et al. 2012), as a consequence favoring metagenomic–agricultural prac-
tices. Additionally metagenomic analyses also depicted several metabolic pro-
cesses, adaptations, and plant growth-promoting characteristics in rice (Sessitsch
8 Metagenomic Approaches in Understanding the Mechanism and Function… 173
2015). Furthermore, the applications can lead to the discovery of new genes, bio-
products, and plant growth-promoting microorganism consortia, useful for under-
standing relevant aspects such as response to stresses (Timmusk et al. 2014) or
dysbiosis (Handelsman 2004; Vayssier-Taussat et al. 2014). Therefore, the amount
of metabolic diversity present in soil to harness novel gene encoding is undeniably
vast which could be explored with the help of metagenomic techniques. Interestingly
the metagenome represents the genomes of uncultured microbes as a rich source for
isolation of many novel genes. In consequence, metagenomics not only provides us
innovative insights into microbial taxonomic/genetic diversity life but also access to
genes (metabolic diversity) producing novel biomolecules (Fig. 8.2).
PGPRs that are difficult to culture are hardly understood and make up a huge part of
soil sequencing techniques that allow us to do more researches. Therefore, the trans-
lational research like metagenomics is highly indispensable to apply these tools for
the benefit of agriculture, particularly by evaluating the effects of decisions on
microbes known to have positive or negative effects on productivity and identify
microbes that are indicators for higher and lower productivity (Souza et al. 2015).
However, preceding works highlighted numerous approaches to harness microbe–
rhizosphere interactions for increased farm productivity (Kaymak 2010; Altomare
and Tringovska 2011; Shen et al. 2013); nevertheless, how these interactions may
perhaps be exploited for biotechnological applications is not completely explored.
Remarkably these rhizosphere–microbe interactions are modulated by numerous
chemical molecules produced by plant roots which in turn communicate with soil
microbes. However, identifying these signal molecules and harnessing them to
improve interaction between beneficial microbes and plant roots can further improve
resource availability. Interestingly one such mechanism explored so far is the
metagenomics of plant microbial signaling that can undoubtedly dole out this ratio-
nale. However, using metagenomics it is now possible to identify mechanisms and
isolate genetic resources for maximizing nutrient cycling and nutrient use efficiency
without cultivating soil microbes (Abhilash et al. 2012).
Nonetheless, utilizing metagenomics to characterize soil microbial communities
facilitates regulation of plant performance through enhanced bioavailability of
nutrients and secretion of bioactive compounds for pest control and hormones to
excite plant growth. In the years to come the plant microbiome may shed light on
solutions to modify plant genetics, function, and ecology for survival in unique
habitats or environmental conditions (Fig. 8.2). Noteworthy, such resolutions may
help to meet rising food demands, curtail land and biodiversity loss, and mitigate the
impacts of climate change. Furthermore understanding the physiological interac-
tions among the microbiomes may allow feasible expression of specific bacterial
traits, viz., nitrogen fixation, phosphate solubilization, ACC deaminase, quorum
sensing, siderophores, chitinases, auxins, gibberellins, or antibiotic production.
Eventually detection of novel genes coding for known activities associated to the
8 Metagenomic Approaches in Understanding the Mechanism and Function… 175
8.9 Conclusions
The advanced tools of metagenomics offer many avenues into a broadened view of
PGPR and their activities in particular. Quantifying the impacts of plant microbi-
ome has advanced our conceptual understanding in plant–microbe metagenomics,
with real-world applications that have the potential to increase crop production dur-
ing enhanced plant trait expression and/or increased utilization of insignificant habi-
tats. Recent advances in metagenomics augmented our overall understanding of
plant–microbe and microbe–microbe interactions in the rhizosphere and also out-
lined quite a few questions for the future. Notably the field of metagenomics, with
reference to soil microbial ecosystems has immense potential, it is hitherto not stud-
ied and explored to a great extent. Interestingly, the discovery of novel PGPR activi-
ties, either by functional screening or sequence information, will add a great deal to
our current understanding of mechanistic variations that exist in PGPR phenotypes.
Additionally, metagenomics can also unravel the gene clusters of biosynthetic-
related genes, synthesis of antimicrobial compounds. Hoping for further advance-
ment of metagenomics in developing successful research and development, PGPR
use will certainly befall a reality and will be helpful to fundamental biochemical and
physiological processes that ascertain the stability and productivity of agroecosys-
tems, thus leading us toward an ideal sustainable agricultural system. The use of
metagenomics in parallel with established or novel molecular approaches will cer-
tainly lead to the discovery of novel mechanisms of PGPR activity, newer types of
PGPR identity, and a spanking new look on the biology and practical applications
of PGPR.
References
Abhilash PC, Powell JR, Singh HB, Singh BK (2012) Plant-microbe interactions: novel applica-
tions for exploitation in multipurpose remediation technologies. Trends Biotechnol 30:416–420
Acosta-Martínez V, Cotton J, Gardner T, Moore-Kucera J, Zak J, Wester D, Cox S (2014)
Predominant bacterial and fungal assemblages in agricultural soils during a record drought/heat
wave and linkages to enzyme activities of biogeochemical cycling. Appl Soil Ecol 84:69–82
176 P.V. Bramhachari et al.
Chhabra S, Brazil D, Morrissey J, Burke JI, O’Gara FN, Dowling D (2013) Characterization of
mineral phosphate solubilization traits from a barley rhizosphere soil functional metagenome.
Microbiol Open 2(5):717–724
Choudhary DK, Sharma KP, Gaur RK (2011) Biotechnological perspectives of microbes in agro-
ecosystems. Biotechnol Lett 33:1905–1910
Daniel R (2005) The metagenomics of soil. Nat Rev Microbiol 3:470e478
Deusch S, Tilocca B, Camarinha-Silva A, Seifert J (2015) News in livestock research-use of omics-
technologies to study the microbiota in the gastrointestinal tract of farm animals. Comput
Struct Biotechnol J 13:55–63
Dey R, Pal KK, Bhatt DM, Chauhan SM (2004) Growth promotion and yield enhancement of pea-
nut (Arachis hypogea L.) by application of plant growth-promoting rhizobacteria. Microbiol
Res 159:371–394
Duan Y, Zhou L, Hall DG, Li W, Doddapaneni H, Lin H et al (2009) Complete genome sequence
of citrus huanglongbing bacterium, ‘Candidatus Liberibacter asiaticus’ obtained through
metagenomics. Mol Plant-Microbe Interact 22:1011–1020
Duan J, Jiang W, Cheng Z, Heikkila JJ, Glick BR (2013) The complete genome sequence of the
plant growth-promoting bacterium Pseudomonas sp. UW4. PLoS One 8:462–469
East R (2013) Microbiome: Soil science comes to life. Nature 501(7468):S18–S19
van Elsas JD, Jansson JK, Trevors JT (2007) Modern soil microbiology. CRC Press. Taylor &
Francis Group, Boca Raton
Faure D, Vereecke D, Leveau JH (2009) Molecular communication in the rhizosphere. Plant Soil
321(1–2):279–303
Fernández-Arrojo L, Guazzaroni ME, López-Cortés N, Beloqui A, Ferrer M (2010) Metagenomic
era for biocatalyst identification. Curr Opin Biotechnol 21(6):725–733
Fierer N, Lauber CL, Ramirez KS, Zaneveld J, Bradford MA, Knight R (2012) Comparative
metagenomic, phylogenetic and physiological analyses of soil microbial communities across
nitrogen gradients. ISME J 6(5):1007e1017
Forchetti G, Masciarelli O, Alemano S, Alvarez D, Abdala G (2007) Endophytic bacteria in sun-
flower (Helianthus annuus L.): isolation, characterization, and production of jasmonates and
abscisic acid in culture medium. Appl Microbiol Biotechnol 76(5):1145–1152
Fujita JM, Kimura N, Sakai A, Ichikawa Y, Hanyu T, Otsuka M (2011) Cloning and heterologous
expression of the Vibrioferrin biosynthetic gene cluster from a marine metagenomic library.
Biosci Biotechnol Biochem 75:2283–2287
Gamalero E, Glick BR (2012) Ethylene and abiotic stress tolerance in plants. In: Environmental
adaptations and stress tolerance of plants in the era of climate change. Springer, New York,
pp 395–412
Glick BR (1995) The enhancement of plant growth by free-living bacteria. Can J Microbiol
41(2):109–117
Glick BR (2012) Plant growth-promoting bacteria: mechanisms and applications. Scientifica.
doi:10.6064/2012/963401
Glick BR (2014) Bacteria with ACC deaminase can promote plant growth and help to feed the
world. Microbiol Res 169:30–39
Haldar S, Sengupta S (2015) Plant-microbe cross-talk in the rhizosphere: insight and biotechno-
logical potential. Open Microbiol J 9:1
Handelsman J (2004) Metagenomics: application of genomics to uncultured microorganisms.
Microbiol Mol Biol Rev 68(4):669–685
Haney CH, Samuel BS, Bush J, Ausubel FM (2015) Associations with rhizosphere bacteria can
confer an adaptive advantage to plants. Nat Plant 1(6)
Hardoim PR, van Overbeek LS, van Elsas JD (2008) Properties of bacterial endophytes and their
proposed role in plant growth. Trends Microbiol 16(10):463–471
Hjort K, Bergström M, Adesina MF, Jansson JK, Smalla K, Sjöling S (2010) Chitinase genes
revealed and compared in bacterial isolates, DNA extracts and a metagenomic library from a
phytopathogen-suppressive soil. FEMS Microbiol Ecol 71:197–207
178 P.V. Bramhachari et al.
Hoff KJ, Tech M, Lingner T, Daniel R, Morgenstern B, Meinicke P (2008) Gene prediction in
metagenomic fragments: a large scale machine learning approach. BMC Bioinform 9(1):1
Hugenholtz P, Goebel BM, Pace NR (1998) Impact of culture-independent studies on the emerging
phylogenetic view of bacterial diversity. J Bacteriol 180(18):4765–4774
Hynes RK, Leung GCY, Hirkala DLM, Nelson LM (2008) Isolation, selection, and characteriza-
tion of beneficial rhizobacteria from pea, lentil, and chickpea grown in western Canada. Can
J Microbial 54:248–258
Ikeda S, Ytow N, Ezura H, Minamisawa K, Miyashita K, Fujimura T (2007) Analysis of Molecular
diversity of bacterial chitinase genes in the maize rhizosphere using culture independent meth-
ods. Microbes Environ 22:71–77
Iovene M, Barone A, Frusciante L, Monti L, Carputo D (2004) Selection for aneuploid potato
hybrids combining a low wild genome content and resistance traits from Solanum commerso-
nii. Theor Appl Genet 109(6):1139–1146
Jahanian A, Chaichi MR, Rezaei K, Rezayazdi K, Khavazi K (2012) The effect of plant growth
promoting rhizobacteria (PGPR) on germination and primary growth of artichoke (Cynara
scolymus). Int J Agric Crop Sci 4:923–929
Jetiyanon K, Kloepper JW (2002) Mixtures of plant growth-promoting rhizobacteria for induction
of systemic resistance against multiple plant diseases. Biol Control 24(3):285–291
Kaymak HC (2010) Potential of PGPR in agricultural innovations. In: Maheshwari DK (ed) Plant
growth and health promoting bacteria. Springer, Berlin, pp 45–79
Kennedy J, Flemer B, Jackson SA, Lejon DP, Morrissey JP, O’Gara F, Dobson AD (2010) Marine
metagenomics: new tools for the study and exploitation of marine microbial metabolism. Mar
Drugs 8(3):608–628
Kristiansson E, Fick J, Janzon A, Grabic R, Rutgersson C, Weijdegard B, Soderstrom H, Larsson
DGJ (2011) Pyrosequencing of antibiotic-contaminated river sediments reveals high levels of
resistance and gene transfer elements. PLoS One 6:e17038
Kumar A, Maurya BR, Raghuwanshi R, Meena VS, Islam MT (2017a) Co-inoculation with
Enterobacter and Rhizobacteria on yield and nutrient uptake by wheat (Triticum aestivum L.)
in the alluvial soil under indo-gangetic plain of India. J Plant Growth Regul. doi:10.1007/
s00344-016-9663-5
Kumar A, Meena VS, Maurya BR, Raghuwanshi R, Bisht JK, Pattanayak A (2017b) Towards the
biological nitrogen fixation and nitrogen management in legume under sustainable agriculture.
Appl Soil Ecol. doi:10.1016/j.apsoil.2017.05.013
Li S et al (2014) Complete genome sequence of Paenibacillus polymyxa SQR-21, a plant growth-
promoting rhizobacterium with antifungal activity and rhizosphere colonization ability.
Genome Announc 2:e00281–e00214
Liu WX et al (2013) Rhizobacteria (Pseudomonas sp. SB) assist phytoremediation of oily-sludge-
contaminated soil by tall fescue (Testuca arundinacea L.) Plant Soil 371:533–542
Lugtenberg B, Kamilova F (2009) Plant-growth-promoting rhizobacteria. Annu Rev Microbiol
63:541–556
Ma Y, Rajkumar M, Luo YM, Freitas H (2011) Inoculation of endophytic bacteria on host and non-
host plants-effects on plant growth and Ni uptake. J Hazard Mater 195:230–237
Martinez-Viveros O, Jorquera MA, Crowley DE, Gajardo G, Mora ML (2010) Mechanisms and
practical considerations involved in plant growth promotion by rhizobacteria. J Soil Sci Plant
Nutr 10:293–319
Masood S, Bano A (2016) Mechanism of potassium solubilization in the agricultural soils by the
help of soil microorganisms. In: Meena VS, Maurya BR, Verma JP, Meena RS (eds) Potassium
solubilizing microorganisms for sustainable agriculture. Springer, New Delhi, pp 137–147.
doi:10.1007/978-81-322-2776-2_10
Meena VS, Maurya BR, Bohra JS, Verma R, Meena MD (2013b) Effect of concentrate manure
and nutrient levels on enzymatic activities and microbial population under submerged rice in
alluvium soil of Varanasi. Crop Res 45(1,2 & 3):6–12
Meena VS, Maurya BR, Verma R, Meena RS, Jatav GK, Meena SK, Meena SK (2013c) Soil
microbial population and selected enzyme activities as influenced by concentrate manure and
inorganic fertilizer in alluvium soil of Varanasi. Bioscan 8(3):931–935
8 Metagenomic Approaches in Understanding the Mechanism and Function… 179
Meena VS, Maurya BR, Bahadur I (2014a) Potassium solubilization by bacterial strain in waste
mica. Bangladesh J Bot 43:235–237
Meena VS, Maurya BR, Verma JP (2014b) Does a rhizospheric microorganism enhance K+ avail-
ability in agricultural soils? Microbiol Res 169:337–347
Meena RS, Meena VS, Meena SK, Verma JP (2015a) The needs of healthy soils for a healthy
world. J Clean Prod 102:560–561
Meena RS, Meena VS, Meena SK, Verma JP (2015b) Towards the plant stress mitigate the agricul-
tural productivity: a book review. J Clean Prod 102:552–553
Meena VS, Maurya BR, Meena RS (2015c) Residual impact of wellgrow formulation and NPK on
growth and yield of wheat (Triticum aestivum L.) Bangladesh J Bot 44(1):143–146
Meena VS, Maurya BR, Verma JP, Aeron A, Kumar A, Kim K, Bajpai VK (2015d) Potassium
solubilizing rhizobacteria (KSR): isolation, identification, and K-release dynamics from waste
mica. Ecol Eng 81:340–347
Meena VS, Meena SK, Verma JP, Meena RS, Ghosh BN (2015e) The needs of nutrient use efficiency
for sustainable agriculture. J Clean Prod 102:562–563. doi:10.1016/j.jclepro.2015.04.044
Meena RS, Bohra JS, Singh SP, Meena VS, Verma JP, Verma SK, Sihag SK (2016b) Towards the
prime response of manure to enhance nutrient use efficiency and soil sustainability a current
need: a book review. J Clean Prod 112(1):1258–1260
Meena SK, Rakshit A, Meena VS (2016c) Effect of seed bio-priming and N doses under varied
soil type on nitrogen use efficiency (NUE) of wheat (Triticum aestivum L.) under greenhouse
conditions. Biocatal Agric Biotechnol 6:68–75
Meena VS, Bahadur I, Maurya BR, Kumar A, Meena RK, Meena SK, Verma JP (2016d) Potassium-
solubilizing microorganism in evergreen agriculture: an overview. In: Meena VS, Maurya BR,
Verma JP, Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture.
Springer, New Delhi, pp 1–20. doi:10.1007/978-81-322-2776-2_1
Meena VS, Meena SK, Bisht JK, Pattanayak A (2016e) Conservation agricultural practices in
sustainable food production. J Clean Prod 137:690–691
Mendes LW, Kuramae EE, Navarrete AA, van Veen JA, Tsai SM (2014) Taxonomical and func-
tional microbial community selection in soybean rhizosphere. ISME J 8(8):1577e1587
Morrell PL, Buckler ES, Ross-Ibarra J (2012) Crop genomics: advances and applications. Nat Rev
Genet 13(2):85–96
Müller CA, Obermeier MM, Berg G (2016) Bioprospecting plant-associated microbiomes.
J Biotechnol 235:171–180
Muyzer G (1999) DGGE/TGGE a method for identifying genes from natural ecosystems. Curr
Opin Microbiol 2(3):317–322
Muyzer G, De Waal EC, Uitterlinden AG (1993) Profiling of complex microbial populations by
denaturing gradient gel electrophoresis analysis of polymerase chain reaction-amplified genes
coding for 16S rRNA. Appl Environ Microbiol 59(3):695–700
Nath D, Maurya BR, Meena VS (2017) Documentation of five potassium- and phosphorus-
solubilizing bacteria for their K and P-solubilization ability from various minerals. Biocatal
Agric Biotechnol 10:174–181
Nikolic B, Schwab H, Sessitsch A (2011) Metagenomic analysis of the 1-aminocyclopropane-1-
carboxylate deaminase gene (acdS) operon of an uncultured bacterial endophyte colonizing
Solanum tuberosum. L Arch Microbiol 193:665–676
Ofek-Lalzar M, Sela N, Goldman-Voronov M, Green SJ, Hadar Y, Minz D (2014) Niche and host-
associated functional signatures of the root surface microbiome. Nat Commun:5, 4950
Öpik M, Moora M, Liira J, Kõljalg U, Zobel M, Sen R (2003) Divergent arbuscular mycorrhizal
fungal communities colonize roots of Pulsatilla spp. in boreal Scots pine forest and grassland
soils. New Phytol 160(3):581–593
Pan Y, Cassman N, de Hollander M, Mendes LW, Korevaar H, Geerts RH, van Veen JA, Kuramae
EE (2014) Impact of long-term N, P, K, and NPK fertilization on the composition and potential
functions of the bacterial community in grassland soil. FEMS Microbiol Ecol 90(1):195–205
Pii Y, Borruso L, Brusetti L, Crecchio C, Cesco S, Mimmo T (2016) The interaction between
iron nutrition, plant species and soil type shapes the rhizosphere microbiome. Plant Physiol
Biochem 99:39–48
180 P.V. Bramhachari et al.
Prakash S, Verma JP (2016) Global perspective of potash for fertilizer production. In: Meena VS,
Maurya BR, Verma JP, Meena RS (eds) Potassium solubilizing microorganisms for sustainable
agriculture. Springer, New Delhi, pp 327–331. doi:10.1007/978-81-322-2776-2_23
Priyadharsini P, Muthukumar T (2016) Interactions between arbuscular mycorrhizal fungi and
potassium-solubilizing microorganisms on agricultural productivity. In: Meena VS, Maurya
BR, Verma JP, Meena RS (eds) Potassium solubilizing microorganisms for sustainable agricul-
ture. Springer, New Delhi, pp 111–125. doi:10.1007/978-81-322-2776-2_8
Qin J, Li R, Raes J, Arumugam M, Burgdorf KS, Manichanh C, …, Mende DR (2010) A human gut
microbial gene catalogue established by metagenomic sequencing. Nature 464(7285):59–65
Raghavendra MP, Nayaka NC, Nuthan BR (2016) Role of rhizosphere microflora in potas-
sium solubilization. In: Meena VS, Maurya BR, Verma JP, Meena RS (eds) Potassium
solubilizing microorganisms for sustainable agriculture. Springer, New Delhi, pp 43–59.
doi:10.1007/978-81-322-2776-2_4
Ramesh R, Joshi AA, Ghanekar MP (2009) Pseudomonads: major antagonistic endophytic bac-
teria to suppress bacterial wilt pathogen, Ralstonia solanacearum in the eggplant (Solanum
melongena L.) World J Microbiol Biotechnol 25(1):47–55
Ramos Solano B, Maicas B, Gutiérrez Mañero J (2008) Physiological and molecular mechanisms
of PGPRs. In: Ahamd I, Pichtel J, Hayat S (eds) En: plant-bacteria interactions. Strategies and
techniques to promote plant growth. Wiley, New York, pp 41–54
Rappé MS, Giovannoni SJ (2003) The uncultured microbial majority. Ann Rev Microbiol 57(1):369–394
Rawat J, Sanwal P, Saxena J (2016) Potassium and its role in sustainable agriculture. In: Meena
VS, Maurya BR, Verma JP, Meena RS (eds) Potassium solubilizing microorganisms for sus-
tainable agriculture. Springer, New Delhi, pp 235–253. doi:10.1007/978-81-322-2776-2_17
Rincon-Florez VA, Carvalhais LC, Schenk PM (2013) Culture-independent molecular tools for
soil and rhizosphere microbiology. Diversity 5(3):581–612
Saha M, Maurya BR, Bahadur I, Kumar A, Meena VS (2016a) Can potassium-solubilising bacteria
mitigate the potassium problems in India? In: Meena VS, Maurya BR, Verma JP, Meena RS
(eds) Potassium solubilizing microorganisms for sustainable agriculture. Springer, New Delhi,
pp 127–136. doi:10.1007/978-81-322-2776-2_9
Saha M, Maurya BR, Meena VS, Bahadur I, Kumar A (2016b) Identification and characterization
of potassium solubilizing bacteria (KSB) from Indo-Gangetic Plains of India. Biocatal Agric
Biotechnol 7:202–209
Sarkar D, Meena VS, Haldar A, Rakshit R (2017) Site-specific nutrient management (SSNM):
a unique approach towards maintaining soil health. In: The adaptive soil management: from
theory to practices, pp 69–88. doi:10.1007/978-981-10-3638-5_3
Schauss K, Focks A, Leininger S, Kotzerke A, Heuer H, Thiele-Bruhn S, ..., Munch JC (2009)
Dynamics and functional relevance of ammonia-oxidizing archaea in two agricultural soils.
Environ Microbiol 11(2):446–456
Schlaeppi K, Dombrowski N, Oter RG, van Themaat EVL, Schulze-Lefert P (2014) Quantitative
divergence of the bacterial root microbiota in Arabidopsis thaliana relatives. Proc Natl Acad
Sci 111(2):585–592
Sessitsch A, Hardoim P, Döring J, Weilharter A, Krause A, Woyke T, ..., Hurek T (2012) Functional
characteristics of an endophyte community colonizing rice roots as revealed by metagenomic
analysis. Mol Plant-Microbe Interactions 25(1):28–36
Sharma A, Shankhdhar D, Shankhdhar SC (2016a) Potassium-solubilizing microorganisms:
mechanism and their role in potassium solubilization and uptake. In: Meena VS, Maurya BR,
Verma JP, Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture.
Springer, New Delhi, pp 203–219. doi:10.1007/978-81-322-2776-2_15
Sharma M, Tarafdar A, Ghosh R (2016b) Use of genomic approaches in understanding the role of
Actinomycetes as PGP in grain legumes. In: Plant growth promoting actinobacteria, Springer,
Singapore, pp 249–262
Shen J, Li C, Mi G, Li L, Yuan L, Jiang R et al (2013) Maximizing root/rhizosphere efficiency to
improve crop productivity and nutrient use efficiency in intensive agriculture of China. J Exp
Bot 64:1181–1192
8 Metagenomic Approaches in Understanding the Mechanism and Function… 181
Shrivastava M, Srivastava PC, D’Souza SF (2016) KSM soil diversity and mineral solubilization,
in relation to crop production and molecular mechanism. In: Meena VS, Maurya BR, Verma JP,
Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture. Springer,
New Delhi, pp 221–234. doi:10.1007/978-81-322-2776-2_16
Simon C, Daniel R (2009) Achievements and new knowledge unraveled by metagenomic
approaches. Appl Microbiol Biotechnol 85(2):265–276
Sindhu SS, Parmar P, Phour M, Sehrawat A (2016) Potassium-solubilizing microorganisms
(KSMs) and its effect on plant growth improvement. In: Meena VS, Maurya BR, Verma JP,
Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture. Springer,
New Delhi, pp 171–185. doi:10.1007/978-81-322-2776-2_13
Singh NP, Singh RK, Meena VS, Meena RK (2015) Can we use maize (Zea mays) rhizobacteria as
plant growth promoter? Vegetos 28(1):86–99. doi:10.5958/2229-4473.2015.00012.9
Singh M, Dotaniya ML, Mishra A, Dotaniya CK, Regar KL, Lata M (2016) Role of biofertil-
izers in conservation agriculture. In: Bisht JK, Meena VS, Mishra PK, Pattanayak A (eds)
Conservation agriculture: an approach to combat climate change in Indian Himalaya. Springer,
Singapore, pp 113–134. doi:10.1007/978-981-10-2558-7_4
Song JY et al (2012) Genome sequence of the plant growth-promoting rhizobacterium Bacillus sp.
strain JS. J Bacteriol 194:3760–3761
Sorensen J, Nicolaisen MH, Ron E, Simonet P (2009) Molecular tools in rhizosphere microbiology
from single-cell to whole-community analysis. Plant Soil 321:483e512
Souza RC, Hungria M, Cantão ME, Vasconcelos ATR, Nogueira MA, Vicente VA (2015)
Metagenomic analysis reveals microbial functional redundancies and specificities in a soil
under different tillage and crop-management regimes. Appl Soil Ecol 86:106–112
Stempfhuber B, Richter-Heitmann T, Regan KM, Kölbl A, Kaul P, Marhan S, Sikorski J, Overmann
J, Friedrich MW, Kandeler E (2015) Spatial interaction of archaeal ammonia-oxidizers and
nitrite-oxidizing bacteria in an unfertilized grassland soil. Front Microbiol 6:1567
Streit WR, Schmitz RA (2004) Metagenomics–the key to the uncultured microbes. Curr Opin
Microbiol 7(5):492–498
Sun L, Qiu F, Zhang X, Dai X, Dong X, Song W (2008) Endophytic bacterial diversity in rice
(Oryza sativa L.) roots estimated by 16S rDNA sequence analysis. Microb Ecol 55(3):415–424
Swarupa V, Pavitra K, Shivashankara KS, Ravishankar KV (2016) Omics-driven approaches in
plant–microbe interaction. In Microbial inoculants in sustainable agricultural productivity,
Springer, India, pp 61–84
Tank N, Saraf M (2010) Salinity-resistant plant growth promoting rhizobacteria ameliorates
sodium chloride stress on tomato plants. J Plant Interact 5:51–58
Teotia P, Kumar V, Kumar M, Shrivastava N, Varma A (2016) Rhizosphere microbes: potassium
solubilization and crop productivity-present and future aspects. In: Meena VS, Maurya BR,
Verma JP, Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture.
Springer, New Delhi, pp 315–325. doi:10.1007/978-81-322-2776-2_22
Timmusk S, El-Daim IAA, Copolovici L, Tanilas T, Kännaste A, Behers L, Nevo E, Seisenbaeva
G, Stenström E, Niinemets Ü (2014) Drought-tolerance of wheat improved by rhizosphere
bacteria from harsh environments: enhanced biomass production and reduced emissions of
stress volatiles. PLoS One 9:e96086
Torsvik V, Sorheim R, Goksoyr J (1996) Total bacterial diversity in soil and sediment communities
a review. J Ind Microbiol 17:170e178
Uhlik O, Leewis MC, Strejcek M, Musilova L, Mackova M, Leigh MB, Macek T (2013) Stable iso-
tope probing in the metagenomics era: a bridge towards improved bioremediation. Biotechnol
Adv 31(2):154–165
Unno Y, Shinano T (2013) Metagenomic analysis of the rhizosphere soil microbiome with respect
to phytic acid utilization. Microbes Environ 28:120–127
Van Emon JM (2015) The omics revolution in agricultural research. J Agric Food Chem 64(1):36–44
Van Oosten VR, Bodenhausen N, Reymond P, Van Pelt JA, Van Loon LC, Dicke M, Pieterse CM
(2008) Differential effectiveness of microbially induced resistance against herbivorous insects
in Arabidopsis. Mol Plant-Microbe Interact 21(7):919–930
182 P.V. Bramhachari et al.
Vayssier-Taussat M, Albina E, Citti C, Cosson J-F, Jacques M-A, Lebrun M-H, Le Loir Y, Ogliastro
M, Petit M-A, Roumagnac P (2014) Shifting the paradigm from pathogens to pathobiome: new
concepts in the light of meta-omics. Front Cell Infect Microbiol 4:29
Velazquez E, Silva LR, Ramírez-Bahena MH, Peix A (2016) Diversity of potassium-solubilizing
microorganisms and their interactions with plants. In: Meena VS, Maurya BR, Verma JP,
Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture. Springer,
New Delhi, pp 99–110. doi:10.1007/978-81-322-2776-2_7
Verma R, Maurya BR, Meena VS (2014) Integrated effect of bio-organics with chemical fertil-
izer on growth, yield and quality of cabbage (Brassica oleracea var capitata). Ind J Agric Sci
84(8):914–919
Verma JP, Jaiswa DK, Meena VS, Meena RS (2015a) Current need of organic farming for enhanc-
ing sustainable agriculture. J Clean Prod 102:545–547
Verma JP, Jaiswal DK, Meena VS, Kumar A, Meena RS (2015b) Issues and challenges about sus-
tainable agriculture production for management of natural resources to sustain soil fertility and
health. J Clean Prod 107:793–794
Verma R, Maurya BR, Meena VS, Dotaniya ML, Deewan P (2017a) Microbial dynamics as influ-
enced by bio-organics and mineral fertilizer in alluvium soil of Varanasi, India. Int J Curr
Microbiol App Sci 6(2):1516–1524
Verma R, Maurya BR, Meena VS, Dotaniya ML, Deewan P, Jajoria M (2017b) Enhancing pro-
duction potential of cabbage and improves soil fertility status of indo-Gangetic plain through
application of bio-organics and mineral fertilizer. Int J Curr Microbiol App Sci 6(3):301–309
Vessey JK (2003) Plant growth promoting rhizobacteria as biofertilizers. Plant Soil 255(2):571–586
van der Vlugt R, Minafra A, Olmos A, Ravnikar M, Wetzel T, Varveri C, Massart S (2015)
Application of next generation sequencing for study and diagnosis of plant viral diseases in
agriculture
Whitman WB, Coleman DC, Wiebe WJ (1998) Prokaryotes: the unseen majority. Proc Natl Acad
Sci 95(12):6578–6583
Wilson SA, Roberts SC (2014) Metabolic engineering approaches for production of biochemicals
in food and medicinal plants. Curr Opin Biotechnol 26:174–182
Woese CR, Fox GE (1977) Phylogenetic structure of the prokaryotic domain: the primary king-
doms. Proc Natl Acad Sci 74(11):5088–5090
Yadav BK, Sidhu AS (2016) Dynamics of potassium and their bioavailability for plant nutrition. In:
Meena VS, Maurya BR, Verma JP, Meena RS (eds) Potassium solubilizing microorganisms for
sustainable agriculture. Springer, New Delhi, pp 187–201. doi:10.1007/978-81-322-2776-2_14
Yasin M, Munir I, Faisal M (2016) Can Bacillus spp. enhance K+ uptake in crop species. In:
Meena VS, Maurya BR, Verma JP, Meena RS (eds) Potassium solubilizing microorganisms for
sustainable agriculture. Springer, New Delhi, pp 163–170. doi:10.1007/978-81-322-2776-2_12
Yergeau E, Sanschagrin S, Maynard C, St-Arnaud M, Greer CW (2014) Microbial expression
profiles in the rhizosphere of willows depend on soil contamination. ISME J 8(2):344–358
Zahedi H (2016) Growth-promoting effect of potassium-solubilizing microorganisms on
some crop species. In: Meena VS, Maurya BR, Verma JP, Meena RS (eds) Potassium sol-
ubilizing microorganisms for sustainable agriculture. Springer, New Delhi, pp 31–42.
doi:10.1007/978-81-322-2776-2_3
Zeyaullah M, Kamli MR, Islam B, Atif M, Benkhayal FA, Nehal M, ..., Ali A (2009)
Metagenomics-An advanced approach for noncultivable micro-organisms. Biotechnol Mol
Biol Rev 4(3):49–54
Zhang Z, Ober U, Erbe M, Zhang H, Gao N, He J, Li J, Simianer H (2014) Improving the accuracy
of whole genome prediction for complex traits using the results of genome wide association
studies. PLoS One 9(3):e93017
Role of Bioinoculants as Plant
Growth-Promoting Microbes 9
for Sustainable Agriculture
Abstract
The green revolution bought amazing consequences in food grain production but
with insufficient concern for agriculture and environmental sustainability. The
efficient biofertilizers are gaining importance in sustaining agriculture. Various
complementing combinations of microbial inoculants for management of major
nutrients are necessary for agriculture sustainability. In such case, microorgan-
isms are useful for biomineralization of bound soil and make nutrients available
to their host and/or its surroundings. Nitrogen and phosphorus are major plant
nutrients which occupy a key place in the balanced use of fertilizer. Leguminous
crop fixes the atmospheric nitrogen by Rhizobium which requires optimum level
of phosphorus in plant tissue. Mineral solubilizers play an important role in seed-
ling setting because more of the tropical soils are phosphate fixing and make it
unavailable to the plants. Phosphate-solubilizing microorganisms (PSMs) that
solubilize bound form of phosphorus and AM fungi act as uptaker of phosphorus
and make it available to the host plants. Microorganisms facilitate plant mineral
nutrition by changing the amounts, concentrations and properties of minerals
available to plants. Because of the various challenges faced in screening, formu-
lation and application, PGPB have yet to fulfil their promise and potential as
commercial inoculants. This chapter focuses on some important information
Keywords
Beneficial bacteria • Inoculant • Plant growth-promoting bacteria (PGPR) •
Sustainable agriculture
9.1 Introduction
In the late 1960s, agriculture system focused on food crop productivity, though
high-yielding varieties (HYVs), agrochemicals, irrigation system and chemical fer-
tilizers were extensively used throughout India. About 50% of the yield growth was
contributed by the use of fertilizer alone. In fact, India is the world’s largest user of
chemical fertilizers, consuming each year ~16% of the world’s N consumption, ~
19% of phosphatic and ~15% of potassic nutrients of the global total (Anonymous
2008). As per the Raghuwanshi (2012) report, out of the ~329 M ha of India’s geo-
graphical area, ~114 M ha are under cultivation. Farmers inoculate the soil with
fertilizers in order to reap a better harvest. Fertilizers function as catalysts for growth
and development of plants by providing proper nutrients. These can be categorized
into chemical and biofertilizer which has its advantages and disadvantages (Tables
9.1 and 9.2).
For the balanced nutrient management for crop growth, these advantages need to
be integrated in order to achieve optimum performance by each type of fertilizer.
High inputs of chemical fertilizers during the last 150 years left soils degraded, pol-
luted and less productive but have also posed severe health and environmental haz-
ards. The use of biofertilizers would solve these issues and make the ecosystem
healthier. It is reported that the ~22 M ha of land are now cultivated organically
which represents <1% of the world’s conventional agricultural production and ~9%
of the total agricultural area (Mishra and Dash 2014).
Essential elements are required for proper growth and functioning of plant. The
shortage of any of these essential nutrients to plant, listed in Table 9.3, can lead to
damage to crop health. Primary macronutrients or micronutrients such as nitrogen,
phosphorus and potassium are required in the greatest quantities, whereas second-
ary macronutrients or micronutrients are needed in smaller quantities, and are found
in adequate quantities in soil, and therefore do not often limit crop growth.
Micronutrients, or trace nutrients, are needed in very small amounts and can be
toxic to plants in excess. Silicon (Si) and sodium (Na) are sometimes considered
essential plant nutrients. Due to their omnipresent nature, they are never in short
supply of these nutrients in soil (Parikh and James 2012).
Biofertilizers play an important role in improving the nutrient supplies and their
availability in plant which includes selective organisms, like bacteria, fungi and
9 Role of Bioinoculants as Plant Growth-Promoting Microbes for Sustainable… 185
Table 9.3 Essential plant nutrient elements and their primary form utilized by plants
Essential plant element Symbol Primary form
Carbon C CO2 (g)
Hydrogen H H2O (l), H+
Oxygen O H2O (l), O2(g)
Mineral elements
Primary macronutrients Nitrogen N NH4+, NO3−
Phosphorus P HPO42−, H2PO4−
Potassium K K+
Calcium Ca Ca2+
Secondary macronutrients Magnesium Mg Mg2+
Sulphur S SO42−
Iron Fe Fe3+, Fe2+
Manganese Mn Mn2+
Zinc Zn Zn2+
Micronutrients Copper Cu Cu2+
Boron B B(OH)3
Molybdenum Mo MoO42−
Chlorine Cl Cl−
Nickel Ni Ni2+
Parikh and James (2012)
algae. These organisms are capable of fixing atmospheric nitrogen and make it
available to the soil for the plant growth. They are eco-friendly, cost-effective and
renewable source of plant nutrients. The biofertilizer plays a vital role in maintain-
ing long-term soil fertility (Mishra et al. 2015). Biofertilizers are economical, eco-
friendly, productive and accessible to small farmers over chemical fertilizers after
the long-term use (Meena et al. 2013a; Bahadur et al. 2014; Maurya et al. 2014; Jat
et al. 2015; Kumar et al. 2016b).
The efficient biofertilizers are the outcome of most advance microbial technology
required to support sustainable and non-polluting agriculture. The biofertilizers
contained living microorganism that, when applied to seed, plant surfaces or soil,
accelerate their microbial processes which result in the availability of nutrients for
easy assimilation by plants (Sharma et al. 2012; Kumar et al. 2015, 2016a; Ahmad
et al. 2016; Meena et al. 2016a; Jha and Subramanian 2016). Since nitrogen is inert
in nature, the plants cannot utilize it from the atmosphere. At ambient temperature
and pressure, diazotrophs mediate nitrogen fixation under enzymatic reaction by a
process known as biological nitrogen fixation (Table 9.4).
9 Role of Bioinoculants as Plant Growth-Promoting Microbes for Sustainable… 187
(a) Vegetative cells: normal photosynthetic cells found under favourable growing
conditions.
(b) Akinetes: climate resistant spores formed under environmental stress
conditions.
(c) Heterocysts: thick-walled cells containing nitrogenase enzyme vital for N2 fixa-
tion. Heterocysts are specialized structures for N2 fixation and able to fix nitro-
gen (N2) into ammonia (NH3+), nitrites (NO2-) or nitrates (NO3-) which can be
utilized by plants.
likely storage structures. Arbuscules are formed inside the plant cells. Nutrients
travel from soil through fungal hyphae to these arbuscules which gradually degener-
ate and provide nutrients to plants. Thus, the association helps in higher uptake of P
and better utilization of other nutrients like N, Cu, Zn, S, etc. (Sharma et al. 2012).
For the growth of plant and soil health, sulphur is an essential nutrient, and the
genus Thiobacillus is an important group of sulphur oxidizer. The application of
sulphur coupled with thiobacilli renders alkali soils fit for cultivation of crops. The
formation of sulphuric acid by Thiobacillus in soil increases the level of soluble P,
K, Ca, Mg, Al and Mn ion. The sulphuric acid produced in the mixture dissolves the
phosphate and thereby enhances phosphorus nutrition of plants (Sharma et al.
2012).
Soil microorganisms have the ability to absorb and mobilize the fixed form of nutri-
ent from trace mineral sources. Silicate bacteria are known to release organic acids
during solubilization (Nath et al. 2017; Sarkar et al. 2017; Verma et al. 2017a). It is
known to dissolve potassium, silica and aluminium. Soil microorganisms for solu-
bilization of silicate minerals are Erwinia, Pseudomonas sp. and Bacillus sp.
(Sharma et al. 2012; Rawat et al. 2016; Yasin et al. 2016; Meena et al. 2016c, e;
Saha et al. 2016a; Masood and Bano 2016).
9 Role of Bioinoculants as Plant Growth-Promoting Microbes for Sustainable… 191
Penicillium citrinum present in soil is known to solubilize manganese from the low-
grade manganese ores from the soil. It produces reductive compounds such as
organic acids which help in solubilizing manganese.
9.4.4 P
lant Growth-Promoting Rhizobacteria (PGPR)
as Biofertilizer
Rhizobacteria aggressively colonize plant roots and multiply and colonize on the
roots in the presence of a competing microflora and enhance plant growth.
1-Aminocyclopropane-1-catrboxylate (ACC) deaminase is the PGPR enzyme that
is able to hydrolyse ACC by lowering ethylene concentration in seedlings; these
PGPR stimulate seedlings’ root length (Sharma et al. 2012). Several PGPR inocu-
lants are commercialized which function as bio-protectants, biofertilizers and bio-
stimulators (Sharma et al. 2012). Several bacterial species such as Pseudomonas,
Azospirillum, Azotobacter, Klebsiella, Enterobacter, Alcaligenes, Arthrobacter,
Burkholderia, Bacillus, Flavobacterium, Microbacterium and Serratia have
reported as the most significant phosphate-solubilizing bacteria (Saharan and Nehra
2011; Bhattacharya and Jha 2012; Yadav and Sidhu 2016; Meena et al. 2016d; Saha
et al. 2016b; Bahadur et al. 2016a).
Piriformospora indica Piriformospora indica is a wide-host root-colonizing endo-
phytic fungus which promotes plant growth and performance. The use of P. indica
as a growth and biomass-stimulating fungus in agriculture is well known because:
(a) Single infection processes are sufficient for the efficient root colonization.
(b) It can survive under adverse environmental conditions, viz. such as cold (<0 °C),
extreme heat (>45 °C), salt and heavy metal-polluted soils and soils with varied
pH values (3.3–9.2).
(c) It promotes micro (e.g. Fe)- and macro (such as P, S and N)-nutrient uptake
from soil.
(d) It confers resistance to root and leaf pathogens through induced systemic resis-
tance (ISR).
(e) It stimulates plant growth and biomass though root exudates.
bacterial strains were used as biofertilizers for nonlegume species with good effi-
cacy results (Verma et al. 2014, 2015b; Meena et al. 2013c, 2014a; Singh et al.
2015). Recently, Hettiarachchi et al. (2014) worked on the fungal-bacterial interac-
tion as biofilmed biofertilizers (BFBFs) in Hevea seedlings and observed that BFBF
application increased root growth of the seedling rubber plants.
Several mechanisms encourage the crop growth by the use of biofertilizers. The
mode of action of commercially available biofertilizers is not always understood;
growth promotion has been identified as the result of indirect or direct mechanisms.
Conversely, direct growth promotion mechanism may provide some compounds
essential for crop growth which stimulate nutrient uptake. Biofertilizers solubilize
the insoluble forms of phosphates like tricalcium, iron and aluminium phosphates
into available forms (Sharma et al. 2016; Verma et al. 2015a; Meena et al. 2013b).
Biofertilizers compose organic matter and help in mineralization in soil. Core
mechanism for mineral phosphate solubilization is the production of organic acids
and acid phosphatases. It is generally accepted that the major mechanism of mineral
phosphate solubilization is the action of organic acids synthesized by soil microor-
ganisms (Shrivastava et al. 2016; Velazquez et al. 2016; Meena et al. 2015c; Sindhu
et al. 2016). The production of organic acids results in acidification of the microbial
cell and its surroundings. The most frequent mean of mineral P solubilization is
gluconic acid and ketogluconic acid. Other organic acids, such as glycolic, oxalic,
malonic and succinic, have also been identified among P solubilizers. Chelating
substances and inorganic acids such as sulphideric, nitric, and carbonic acid are
considered as other mechanisms for P solubilization. However, the efficacy and
their contributions to P release in soils seem to be less than organic acid production.
In recent years, the use of artificially produced inoculum of mycorrhizal fungi has
increased its significance due to their varied role in plant growth and yield and resis-
tance against climatic and edaphic stresses, pathogens and pests (Meena et al.
2014b; Singh et al. 2016).
9.6.1 Carriers
The delivery vehicle of live microorganisms from the factory to the field is called
carrier (Trevors et al. 1992). A good carrier should therefore possess as much of the
following properties:
In case the inoculant is used as seed coating, the carrier shall assure the survival
of the PGPM on the seed. A good carrier should have one essential characteristic:
the capacity to deliver the right number of viable cells in good physiological condi-
tion at the right time (Trevors et al. 1992; Meena et al. 2015d).
Peat formulations have been the carriers of choice and are the most commonly
used in the rhizobia inoculation industry (Kaljeet et al. 2011). Peat inoculant
applied to the seed as slurry is the most commonly used method to inoculate grain
legumes with rhizobia (e.g. Bradyrhizobium sp., Mesorhizobium sp., Rhizobium
sp., etc.). Peat slurry inoculants are made using finely milled peat that has been
sterilized by gamma irradiation, and these sterilized inoculants can support high
concentrations of rhizobia, generally 109–1010 cells g−1 peat at manufacture (Hartley
et al. 2005).
A popular method for storing and delivering microorganisms through liquid formu-
lations is water-in-oil emulsions (Vandergheynst et al. 2006). This formulation
slows down water evaporation as the oil traps the water around the organism which
is particularly beneficial for organisms that are sensitive to desiccation. Thickening
the oil phase using hydrophobic silica nanoparticles significantly reduced cell sedi-
mentation and improved cell viability during storage (Vandergheynst et al. 2007).
9 Role of Bioinoculants as Plant Growth-Promoting Microbes for Sustainable… 195
Recently, a new process named PGSS (particles from gas saturated solutions), based
on the application of supercritical fluid properties, is used which is carried out at
low temperatures and uses carbon dioxide as a supercritical fluid. Another new tech-
nology is proposing the exploitation of the natural production of bacterial biofilms
as a possible carrier. Application of a biofilmed inoculant containing a fungal rhizo-
bia consortium significantly increased N2 fixation in soybean compared to a tradi-
tional rhizobium inoculant (Jayasinghearachchi and Seneviratne 2004).
Interestingly, beneficial endophytes were observed to produce higher acidity and
plant growth-promoting hormones than their mono- or mixed cultures with no bio-
film formation (Bandara et al. 2006). Wheat seedlings inoculated with biofilm-
producing bacteria exhibited an increased yield in moderate saline soils (Ashraf
et al. 2004). Inocula made with biofilms were shown to allow their rhizobia survive
at high salinity (400 mM NaCl) by 105-fold compared to rhizobial monocultures
(Seneviratne et al. 2008). Bio-nanotechnology applications which employ nanopar-
ticles made of inorganic or organic materials could also provide new avenues for the
development of carrier-based microbial inocula (Malusa et al. 2012).
9.10 C
ommercially Available Biofertilizer in India and Its
Application
9.11 Conclusions
Microbial inoculants have long been incorporated into field practices worldwide.
The demand for natural biofertilizers is rising steadily in all parts of the world.
Public awareness about the environment is increasing, while the apprehension about
pollution and health hazards due to synthetic chemicals especially in rich countries
is growing. Significant advances have been made by some Asian countries in the
development and use of biofertilizers, but their potential remains largely underuti-
lized due to the difference in efforts and experiences in different countries. Peat
Table 9.5 Microbial biofertilizer available in India
196
(continued)
9 Role of Bioinoculants as Plant Growth-Promoting Microbes for Sustainable…
197
Table 9.5 (continued)
198
(continued)
199
Table 9.5 (continued)
200
formulations have been developed into effective carriers which are accepted world-
wide, but their development has almost reached its limits. It is quite early to declare
these carriers as potentially universal due to the lack of information about new
developments from research institute. Special attention should be paid to the needs
and constraints of developing countries that need easy-to-use and inexpensive for-
mulations as agriculture in developed countries is the major promoter of microbial
inoculants that are environmentally friendly. For the future, more research should be
focused on the development of better and more economical feasible, synthetic inoc-
ulant carriers, while sustaining peat-based inoculant production for agriculture.
References
Ahmad M, Nadeem SM, Naveed M, Zahir ZA (2016) Potassium-solubilizing bacteria and their
application in agriculture. In: Meena VS, Maurya BR, Verma JP, Meena RS (eds) Potassium
solubilizing microorganisms for sustainable agriculture. Springer, New Delhi, pp 293–313.
doi:10.1007/978-81-322-2776-2_21
Amer GA, Utkhede RS (2000) Development of formulations of biological agents for management
of root rot of lettuce and cucumber. Can J Microbiol 46:809–816
Amutha R, Karunakaran S, Dhanasekaran S, Hemalatha K, Monika R, Shanmugapriya P,
Sornalatha T (2014) Isolation and mass production of biofertilizer (Azotobacter and phospho-
bacter). Int J Lat Res Sci Tech 3(1):79–81
Anonymous (2008) Data on bio-fertilizer production and demand, maintained by Department of
Fertilizers. Ministry of Chemical and Fertilizers, New Delhi. Based upon information received
from State Govt Regional Centers/NGO
Ashraf M, Hasnain S, Berge O, Mahmood T (2004) Inoculating wheat seedlings with exopolysac-
charide -producing bacteria restricts sodium uptake and stimulates plant growth under salt
stress. J Food Sci 71(3):89–99
Bahadur I, Meena VS, Kumar S (2014) Importance and application of potassic biofertilizer in
Indian agriculture. Int Res J Biol Sci 3:80–85
Bahadur I, Maurya BR, Kumar A, Meena VS, Raghuwanshi R (2016a) Towards the soil sustain-
ability and potassium-solubilizing microorganisms. In: Meena VS, Maurya BR, Verma JP,
Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture. Springer,
New Delhi, pp 225–266. doi:10.1007/978-81-322-2776-2_18
Bahadur I, Maurya BR, Meena VS, Saha M, Kumar A, Aeron A (2016b) Mineral release dynamics
of tricalcium phosphate and waste muscovite by mineral-solubilizing rhizobacteria isolated
from indo-gangetic plain of India. Geomicrobiol J. doi:10.1080/01490451.2016.1219431
Bahadur I, Maurya BR, Meena VS, Saha M, Kumar A, Aeron A (2017) Mineral release dynamics
of tricalcium phosphate and waste muscovite by mineral-solubilizing rhizobacteria isolated
from indo-gangetic plain of India. Geomicrobiol J. doi:10.1080/01490451.2016.1219431
Bandara WMMS, Seneviratne G, Kulasooriya SA (2006) Interactions among endophytic bacteria
and fungi: effects and potentials. J Biosci 31(5):645–650
Baset Mia MA, Shamsuddin ZH (2010) Rhizobium as a crop enhancer and biofertilizer for
increased cereal production. Afr J Biotechnol 9(37):6001–6009
Bashan Y (1998) Inoculants of plant growth-promoting bacteria for use in agriculture. Biotech
Adv 16:729–770
Bhattacharyya PN, Jha DK (2012) Plant growth-promoting rhizobacteria (PGPR): emergence in
agriculture. World J Microbiol Biotechnol 28:1327–1350
Bora T, Ozaktan H, Gore E, Aslan E (2004) Biological control of Fusarium oxysporum f. sp.
Melonis by wettable powder formulations of the two strains of Pseudomonas putida.
J Phytopathol 152:471–475
202 T. Parween et al.
Mishra S, Singh A, Keswani C, Saxena A, Sarma BK, Singh HB (2015) Harnessing plant-microbe
interactions for enhanced protection against phytopathogens. In: Plant microbes symbiosis:
applied facets. Springer India, p 111–125
Murumkar DR, Nalawade S V, Indi DV, Pawar SM (2016) Response of sugarcane seed plot to
microbial inoculation by Gluconacetobacter diazotrophicus and phosphate-solubilizing bac-
teria. Sugar Tech 6:1–7
Muthukumarasamy R, Revathi G, Lakshminarasimhan C (1999) Diazotrophic associations in sug-
arcane cultivation in South India. Trop Agric 76:171–178
Nath D, Maurya BR, Meena VS (2017) Documentation of five potassium- and phosphorus-
solubilizing bacteria for their K and P-solubilization ability from various minerals. Biocatal
Agric Biotechnol 10:174–181
Parewa HP, Yadav J, Rakshit A, Meena VS, Karthikeyan N (2014) Plant growth promoting rhizo-
bacteria enhance growth and nutrient uptake of crops. Agric Sustain Dev 2(2):101–116
Parikh SJ, James BR (2012) Soil: the foundation of agriculture. Nat Educ Knowl 3(10):2
Partha N, Sivasubramanian V (2006) Recovery of chemicals from press mud – a sugar industry
waste. Indian Chem Eng 48(3):161–163
Prakash S, Verma JP (2016) Global perspective of potash for fertilizer production. In: Meena VS,
Maurya BR, Verma JP, Meena RS (eds) Potassium solubilizing microorganisms for sustainable
agriculture. Springer, New Delhi, pp 327–331. doi:10.1007/978-81-322-2776-2_23
Priyadharsini P, Muthukumar T (2016) Interactions between arbuscular mycorrhizal fungi and
potassium-solubilizing microorganisms on agricultural productivity. In: Meena VS, Maurya
BR, Verma JP, Meena RS (eds) Potassium solubilizing microorganisms for sustainable agricul-
ture. Springer, New Delhi, pp 111–125. doi:10.1007/978-81-322-2776-2_8
Raghavendra MP, Nayaka NC, Nuthan BR (2016) Role of rhizosphere microflora in potas-
sium solubilization. In: Meena VS, Maurya BR, Verma JP, Meena RS (eds) Potassium
solubilizing microorganisms for sustainable agriculture. Springer, New Delhi, pp 43–59.
doi:10.1007/978-81-322-2776-2_4
Raghuwanshi R (2012) Opportunities and challenges to sustainable agriculture in India. NEBIO
3(2):78–86
Rawat J, Sanwal P, Saxena J (2016) Potassium and its role in sustainable agriculture. In: Meena
VS, Maurya BR, Verma JP, Meena RS (eds) Potassium solubilizing microorganisms for sus-
tainable agriculture. Springer, New Delhi, pp 235–253. doi:10.1007/978-81-322-2776-2_17
Reid WS, Liptay A, Nicholls CF, Marriage PB (1983) A plug-mix planter attachment for dispens-
ing a charcoal vermiculite mixture to protect emerging seedlings from herbicide toxicity. Can
J Plant Sci 63:567–571
Saha M, Maurya BR, Bahadur I, Kumar A, Meena VS (2016a) Can potassium-solubilising bacteria
mitigate the potassium problems in India? In: Meena VS, Maurya BR, Verma JP, Meena RS
(eds) Potassium solubilizing microorganisms for sustainable agriculture. Springer, New Delhi,
pp 127–136. doi:10.1007/978-81-322-2776-2_9
Saha M, Maurya BR, Meena VS, Bahadur I, Kumar A (2016b) Identification and characterization
of potassium solubilizing bacteria (KSB) from indo-gangetic plains of India. Biocatal Agric
Biotechnol 7:202–209
Saharan BS, Nehra V (2011) Plant growth promoting rhizobacteria: a critical review. Life Sci Med
Res 21. http://astonjournals.com/lsmr
Saravanakumar D, Vijayakumar C, Kumar N, Samiyappan R (2007a) PGPR induced defense
responses in tea plants against blister blight disease. Crop Protect 26:556–565
Saravanakumar D, Harish S, Loganathan M, Vivekananthan R, Rajendran L, Samiyappan R
(2007b) Rhizobacterial bioformulation for the effective management of Macrophomina root
rot in mung bean. Arch Phytopathol Plant Protect 40(5):323–337
Sarkar D, Meena VS, Haldar A, Rakshit R (2017) Site-specific nutrient management (SSNM):
a unique approach towards maintaining soil health. In: The Adaptive soil management: from
theory to practices, pp 69–88. doi:10.1007/978-981-10-3638-5_3
9 Role of Bioinoculants as Plant Growth-Promoting Microbes for Sustainable… 205
Seneviratne G, Zavahir JS, Bandara WMMS, Weerasekara MLMW (2008) Fungal-bacterial bio-
films: their development for novel biotechnological applications. World J Microbiol Biotechnol
24(6):739–743
Sharma P, Kaur N, Gargi K (2012) Revitalizing soil health with biofertilizers. Biofert News Lett
20(2)
Sharma A, Shankhdhar D, Shankhdhar SC (2016) Potassium-solubilizing microorganisms:
mechanism and their role in potassium solubilization and uptake. In: Meena VS, Maurya BR,
Verma JP, Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture.
Springer, New Delhi, pp 203–219. doi:10.1007/978-81-322-2776-2_15
Shrivastava M, Srivastava PC, D’Souza SF (2016) KSM soil diversity and mineral solubilization,
in relation to crop production and molecular mechanism. In: Meena VS, Maurya BR, Verma JP,
Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture. Springer,
New Delhi, pp 221–234. doi:10.1007/978-81-322-2776-2_16
Sindhu SS, Parmar P, Phour M, Sehrawat A (2016) Potassium-solubilizing microorganisms
(KSMs) and its effect on plant growth improvement. In: Meena VS, Maurya BR, Verma JP,
Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture. Springer,
New Delhi, pp 171–185. doi:10.1007/978-81-322-2776-2_13
Singh NP, Singh RK, Meena VS, Meena RK (2015) Can we use maize (Zea mays) rhizobacteria as
plant growth promoter? Vegetos 28(1):86–99. doi:10.5958/2229-4473.2015.00012.9
Singh M, Dotaniya ML, Mishra A, Dotaniya CK, Regar KL, Lata M (2016) Role of biofertil-
izers in conservation agriculture. In: Bisht JK, Meena VS, Mishra PK, Pattanayak A (eds)
Conservation agriculture: an approach to combat climate change in Indian Himalaya. Springer,
Singapore, pp 113–134. doi:10.1007/978-981-10-2558-7_4
Stephens JHG, Rask HM (2000) Inoculant production and formulation. Field Crops Res
65:249–258
Teotia P, Kumar V, Kumar M, Shrivastava N, Varma A (2016) Rhizosphere microbes: potassium
solubilization and crop productivity-present and future aspects. In: Meena VS, Maurya BR,
Verma JP, Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture.
Springer, New Delhi, pp 315–325. doi:10.1007/978-81-322-2776-2_22
Trevors JT, van Elsas JD, Lee H, vanOverbeek LS (1992) Use of alginate and other carriers for
encapsulation of microbial cells for use in soil. Microb Releases 1:61–69
Vahed HS, Shahinrokhsar P, Heydarnezhad F (2012) Performance of phosphate solubilizing bac-
teria for improving growth and yield of rice (Oryza sativa L.) in the presence of phosphorous
fertilizer. Int J Agric Crop Sci 4(17):1228–1232
Vaid SK, Kumar B, Sharma A, Shukla AK, Sivastava PC (2014) Effect of zinc solubilizing bacteria
on growth promotion and zinc nutrition of rice. J Soil Sci Plant Nutr 13(4):889–910
Vandergheynst JS, Scher H, Hong-Yun G (2006) Design of formulations for improved biological
control agent viability and sequestration during storage. Ind Biotechnol 2(3):213–219
Vandergheynst JS, Scher HB, Guo HY, Schultz DL (2007) Water-in-oil emulsions that improve
the storage and delivery of the biolarvacide Lagenidium giganteum. BioControl 52(2):207–229
Vassileva M, Serrano M, Bravo V, Jurado E, Nikolaeva I, Martos V, Vassilev N (2010)
Multifunctional properties of phosphate-solubilizing microorganisms grown on agro-industrial
wastes in fermentation and soil conditions. Appl Microbiol Biotechnol 85(5):1287–1299
Velazquez E, Silva LR, Ramírez-Bahena MH, Peix A (2016) Diversity of potassium-solubilizing
microorganisms and their interactions with plants. In: Meena VS, Maurya BR, Verma JP,
Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture. Springer,
New Delhi, pp 99–110. doi:10.1007/978-81-322-2776-2_7
Verma R, Maurya BR, Meena VS (2014) Integrated effect of bio-organics with chemical fertilizer
on growth, yield and quality of cabbage (Brassica oleracea var capitata). Indian J Agric Sci
84(8):914–919
Verma JP, Jaiswa DK, Meena VS, Meena RS (2015a) Current need of organic farming for enhanc-
ing sustainable agriculture. J Clean Prod 102:545–547
206 T. Parween et al.
Verma JP, Jaiswal DK, Meena VS, Kumar A, Meena RS (2015b) Issues and challenges about sus-
tainable agriculture production for management of natural resources to sustain soil fertility and
health. J Clean Prod 107:793–794
Verma R, Maurya BR, Meena VS, Dotaniya ML, Deewan P (2017a) Microbial dynamics as
influenced by bio-organics and mineral fertilizer in alluvium soil of Varanasi. India Int J Curr
Microbiol App Sci 6(2):1516–1524
Verma R, Maurya BR, Meena VS, Dotaniya ML, Deewan P, Jajoria M (2017b) Enhancing pro-
duction potential of cabbage and improves soil fertility status of indo-gangetic plain through
application of bio-organics and mineral fertilizer. Int J Curr Microbiol App Sci 6(3):301–309
Vidhyasekaran P, Muthamilan M (1995) Development of formulations of Pseudomonas fluores-
cens for control of chickpea wilt. Plant Dis 79:782–786
Yadav BK, Sidhu AS (2016) Dynamics of potassium and their bioavailability for plant nutrition. In:
Meena VS, Maurya BR, Verma JP, Meena RS (eds) Potassium solubilizing microorganisms for
sustainable agriculture. Springer, New Delhi, pp 187–201. doi:10.1007/978-81-322-2776-2_14
Yasin M, Munir I, Faisal M (2016) Can Bacillus spp. enhance K+ uptake in crop species. In: Meena
VS, Maurya BR, Verma JP, Meena RS (eds) Potassium solubilizing microorganisms for sus-
tainable agriculture. Springer, New Delhi, pp 163–170. doi:10.1007/978-81-322-2776-2_12
Zahedi H (2016) Growth-promoting effect of potassium-solubilizing microorganisms on
some crop species. In: Meena VS, Maurya BR, Verma JP, Meena RS (eds) Potassium sol-
ubilizing microorganisms for sustainable agriculture. Springer, New Delhi, pp 31–42.
doi:10.1007/978-81-322-2776-2_3
Microbial Diversity of Tropical Andean
Soils and Low-Input Sustainable 10
Agriculture Development
Abstract
The Tropical Andes spans over 1,540,000 km2 from western Venezuela to north-
ern Chile and Argentina and is currently considered the most biologically rich
and diverse region on Earth. The rural populations from the Andean highlands
depend largely on agriculture and related activities for their livelihoods. In spite
of a long history of agriculture in the region, crop production in these highlands
is constrained by adverse natural factors such as low soil fertility, rugged ter-
rains, water deficit, and cold climates. In the last decades, a better understanding
of the functioning of microbial communities in agricultural soils has led to the
development of technologies that could alleviate some of these natural con-
straints and increase productivity sustainably. A promising approach to take
advantage of the functioning of these microbial communities is the development
of bioinoculants, products containing live or latent cells of agriculturally benefi-
cial strains of microorganisms. When applied to seeds or soils, these bioinocu-
lants boost the populations of beneficial microorganisms in the rhizosphere and
accelerate certain microbial processes that result in increased nutrient acquisition
by plants and in augmented antagonism to pathogens. A key step in the process
of developing this kind of bioinoculants is the study of the indigenous microbial
Keywords
Bioinoculants • Plant growth-promoting microorganisms • Tropical Andes •
Sustainable agriculture • Biofertilization • Biocontrol
10.1 Introduction
The Tropical Andes is a vast and diverse territory spanning north to south from
western Venezuela to northern Chile and Argentina, next to the Pacific coast of
South America. Home to ~70 million people – almost half of the population of the
Andean countries – the region represents a melting pot of cultures in which both
ancient and modern traditions hybridize (Sandor and Eash 1995; Stanish 2007)
(Fig. 10.1a). The livelihoods of the people living in the Tropical Andes are strongly
linked to agriculture. Indeed, mixtures of indigenous and introduced crops are cul-
tivated by Andean farmers to satisfy national food demands, in agricultural produc-
tion systems mostly dominated by smallholders and family farmers (Fonte et al.
2012) (Fig. 10.1b, c).
During the last decades, a tremendous demographic growth in the region has led
to unprecedented pressures on natural resources and to an increase of rural poverty
and hunger. To satisfy food demands of this growing population, the use of natural
resources has been intensified in an attempt to achieve higher productivities. Alas,
agriculture in the Andean highlands faces several challenges that limit productivity
gains. For instance, besides lacking the financial resources needed to afford fertil-
izers, pesticides, irrigation systems, machinery, and/or improved crop varieties,
small Andean farmers must frequently deal with harsh climate conditions and
nutrient-limited soils (Fonte et al. 2012). These production conditions have caused
many farmer families to suffer food shortages of varying degrees, exacerbating the
poverty-resource degradation-scarcity-poverty cycle (Jodha and Shrestha 1993;
Meena et al. 2013a; Bahadur et al. 2014; Maurya et al. 2014; Jat et al. 2015; Kumar
et al. 2015, 2016b; Ahmad et al. 2016).
It is, therefore, imperative to explore new options aimed at increasing the pro-
ductivity and carrying capacity of farms, in order to improve the livelihoods of
Andean farmers and allowing them to compete favorably in the modern world with-
out causing further environmental degradation (Partap 1999). Among these options,
the Food and Agriculture Organization of the United Nations identified the follow-
ing: (1) cultivation of high-value cash crops (like fruits and vegetables), (2) adop-
tion of sustainable farming practices (which avoid the abuse and misuse of synthetic
10 Microbial Diversity of Tropical Andean Soils and Low-Input Sustainable… 209
Fig. 10.1 (a) The Tropical Andes Region. ©2005 CI/CABS (Reproduced with permission of
Critical Ecosystem Partnership Fund). Available at: http://www.cepf.net/where_we_work/regions/
south_america/tropical_andes/Pages/default.aspx. (b) Small farms in the Andean region of
Ecuador (Highlands or Páramos). Copyright: Luis Andrés Yarzábal (Reproduced with permission)
agrochemicals and fertilizers), and (3) rescue of native species, varieties, and breeds
known as neglected crops, which have played an important role in the human diet
and traditional cultures for millennia (FAO Reports 2003, 2013).
A complementary and promising strategy for improving the agricultural produc-
tivity of Andean farms – without serious social, economic, or environmental
impacts – is the proper use of biotechnological products, such as bioinoculants (also
termed microbial inoculants). A bioinoculant can be defined as a product containing
individual strains or consortia of known microbes that have plant growth-promoting
effects (plant growth-promoting microorganisms; PGPM) when added directly to
the soil or as a seed coating during planting (Owen et al. 2014). Among currently
used bioinoculants, we can mention commercial formulations containing nitrogen-
fixing bacteria (NFB or N-fixers), phosphate solubilizing bacteria (PSB), and/or
mycorrhizal fungi (MF).
The use of such kind of bioinoculants has allowed to increase agricultural pro-
ductivity and sustainability worldwide, without harming the environment (Fuentes-
Ramírez and Caballero-Mellado 2006; Singh et al. 2011a; Velivelli et al. 2014a),
thus becoming a real alternative to synthetic agrochemicals. Contrary to the general
opinion, adoption of bioinoculants by the local farmers in underdeveloped coun-
tries, particularly in the Andean region, can be achieved successfully and perceived
favorably if adequately planned and addressed. More importantly, in the long-term,
210 L.A. Yarzábal et al.
the use of bioinoculants, together with appropriate land management and selection
of suitable cultivars, may positively impact the livelihoods of individual farmers and
their families (Meena et al. 2016a, b; Parewa et al. 2014; Prakash and Verma 2016;
Jha and Subramanian 2016; Kumar et al. 2016a).
The Tropical Andes hold an old and rich tradition in agriculture. Alongside Central
America, the Tropical Andes are considered the cradle of agriculture in the Americas,
with archeological evidence of farming dating back to ~9000 years before the pres-
ent (Dillehay et al. 2007; Piperno 2011). Dominated mainly by the influence of the
Inca Empire (Tawantinsuyu) formed by the assimilation of different peoples, just
prior to the arrival of European conquistadores, agriculture in the Tropical Andes
exhibited an advanced degree of ecological specialization and technological inten-
sification that allowed cultivation on steep mountain slopes. Inca agriculture took
advantage of the natural altitudinal gradient formed in the slopes of the Andes, to
cultivate crops with different temperature requirements in a highly organized fash-
ion that allowed feeding a population of about eight million at its height (Borsdorf
and Stadel 2015).
Water and soil management was an important issue in Inca agriculture, and sev-
eral strategies such as long-distance irrigation canals (acequias), raised bed (waru-
waru), sunken fields (qochas), and field terraces (andenes) were used to conserve
these resources (Borsdorf and Stadel 2015). Incan farmers managed also practices
such as intercropping (chacras), crop rotation, and planned fallows to maintain and
restore soil fertility and point manage the incidence of pests (Borsdorf and Stadel
2015; Pestalozzi 2000). To date, many of these traditional agricultural practices are
still commonly used among Andean farmers.
Modern agriculture in the Tropical Andes is hard to define because of the diver-
sity of technologies and traditions it is made of. Different criteria could be used to
classify these production systems (Borsdorf and Stadel 2015; Mulligan et al. 2009);
however, by integrating several of these criteria, three broad categories can be iden-
tified, namely, (1) smallholder commercial and subsistence farms, (2) large-scale
commercial export-oriented farms, and (3) managed pastures and rangelands. These
three systems are roughly distributed along an altitudinal gradient, with large farms
located at lower altitudes, where soils are more fertile, and small farms and pastures
located at higher altitudes, on marginally fertile lands. These systems also differ
among themselves in terms of their demand for external inputs, their impact on
natural resources, their productivity and their level of technological intensification
(Priyadharsini and Muthukumar 2016; Kumar et al. 2017; Meena et al. 2015a;
Jaiswal et al. 2016).
By large, agriculture in the Tropical Andes is dominated by smallholder com-
mercial and subsistence farms. These farms concentrate around towns and cities, in
the valleys, on hillslopes, and in the high plateaus (altiplanos) up to ~4200 m asl
(Borsdorf and Stadel 2015; Mulligan et al. 2009) (Fig. 10.1b, c). Agricultural
10 Microbial Diversity of Tropical Andean Soils and Low-Input Sustainable… 211
Table 10.1 Commercially cultivated crops in Tropical Andean agricultural production systems
Smallholders farms Large-scale farms Pastures and rangelands
Native cultivars Introduced cultivars Forages Livestock
Corn Tomato Rose Kikuyu Cattle
Bean Peppers Gypsophila Ryegrass Sheep
Potato Broccoli Hypericum Alfalfa Goats
Squash Cauliflower Tomato Clover Llamas
Quinoa Cabbage Broccoli Alpacas
Amaranth Radish Alcachofa Vicuñas
Oca Spinach Sugarcane
Mashwa Onion
Ullucus Lettuce
Chili pepper Carrot
Coffee
production in these farms is devoted to local and national markets and to self-
consumption by farmer families. In general, a greater share of agricultural produc-
tion is used for self-consumption in upland communities that are farther away from
urban centers (Mulligan et al. 2009).
Although productivity in these smallholder farms is low compared to large-scale
commercial farms, they supply most of the national demand of fresh fruits and veg-
etables in their corresponding countries and, thus, have a significant socioeconomic
impact (Devenish and Gianella 2012). Crops cultivated in these small farms are
highly diverse and represent a mixture of native and introduced crops (Table 10.1),
which are cultivated using a combination of traditional practices and modern tech-
nology. However, low productivity, inappropriate adoption of production practices
and technologies, and lack of research oriented to understand and improve these
production systems threaten the sustainability and environmental health of these
small farms (Bojaca et al. 2010; Cole et al. 2011).
In contrast with smallholder farms, large-scale commercial and export-oriented
farms in the Tropical Andes are concentrated almost uniquely in the warmer valleys.
The main crops cultivated in these systems are vegetables (e.g., broccoli, artichoke,
tomato, peppers, lettuce), cut flowers (e.g., roses, Gypsophila, Hypericum), and sug-
arcane. The production of these systems is destined in part to supply national super-
market chains, but the majority of it is exported to foreign markets. These systems
exhibit high productivity and intensification and contribute to rural economies by
providing rural employment and generating external revenue; however, they gener-
ate a larger environmental impact than small farms and increase the dependence on
external inputs (Mulligan et al. 2009). In the last decades, market preferences and
environmental regulations have promoted the introduction, in many of these large-
scale operations, of practices that reduce the negative impacts they generate on the
environment (Raghavendra et al. 2016; Zahedi 2016; Meena et al. 2015b, f; Rawat
et al. 2016; Yasin et al. 2016).
The third type of agricultural production system in the Tropical Andes is the
managed pastures and rangelands for livestock grazing. These systems can be found
212 L.A. Yarzábal et al.
typically at greater elevations (>3000 m asl) on mountain slopes and high plateaus
up to ~4500 m asl (Borsdorf and Stadel 2015). The most common species managed
in these systems are dairy cattle, sheep, goats, and Andean cameloids (llamas,
alpacas, and vicuñas) (Borsdorf and Stadel 2015). Among these species, dairy cattle
are the most commercially important species in the Tropical Andes. Both large and
small dairy cattle farmers (2–30 cattle) with different levels of technological inten-
sification coexist in the Tropical Andes (Aubron et al. 2009). Most of the dairy cattle
are fed on rangelands or on crop residues, with few farmers actually establishing
and managing pastures and fodder crops (Aubron et al. 2009).
In certain parts of the Peruvian and Bolivian Andes, elaborated long rotations
(~12 years) of crops (potato, quinoa, oats, barley) and fallows are used (Pestalozzi
2000). Common fodder crops used by dairy cattle ranchers are kikuyu, ryegrass,
clover, and alfalfa, whereas common breeds include Holstein, Brown Swiss, and
Criollos (Aubron et al. 2009). Historically, cattle farming in the Tropical Andes has
been one of the main drivers for the deforestation and ecosystem degradation since
native forests are normally cleared to create new rangelands and later abandoned
when productivity starts to decrease (Lozada 2006). These abandoned rangelands
are difficult to recover either to reestablish commercial production or for restoring
the original ecosystem (Posada et al. 2000).
Agricultural production systems in the Tropical Andes are also strongly influ-
enced by local natural and cultural factors. Depending on their location and the
scale of operations, different systems show different levels of sophistication in
terms of cultural practices and inclusion of technology, which in turn affect the
sustainability of these systems. Being the most numerous, a better understanding of
the functioning of smallholder production systems, as well as their optimization and
sustainable intensification, would have a greater overall impact on the sustainability
of agriculture in the Tropical Andes in the face of emerging challenges (Devenish
and Gianella 2012; Fonte et al. 2012; Haller 2014; Mulligan et al. 2010).
The Tropical Andes exhibits a high diversity of soil types (Gardi et al. 2014), whose
detailed description goes beyond the scope of this chapter. However, considering
that the most vulnerable population (small farmers) are established in the Andean
highlands (over 2500 m asl) (Fonte et al. 2012) and that they depend mostly on
agriculture for their livelihoods, it is of the utmost importance to describe some
characteristics of the most representative soils in this region.
Soils in the Tropical Andes can be defined as those located between the Tropic of
Cancer (23°26′16″N) and the Tropic of Capricorn (23°26′16″) (Gardi et al. 2014)
and above 2500 m asl. Alpine soils dominate above the tree line and, among them,
páramo soils stand out, with Andosols as the most extended soil type (Poulenard
and Podwojewski 2006; Podwojewski and Poulenard 2006). Nevertheless, Histosols,
Leptosols, and Regosols are also present (Buytaert et al. 2011), mainly in large
10 Microbial Diversity of Tropical Andean Soils and Low-Input Sustainable… 213
extensions at the Puna highland region of the Central Andes (Gardi et al. 2014;
Ochoa-Tocachi et al. 2016).
Andosols are, in essence, volcanic ash soils (Shoji et al. 1993). These soils
exhibit different features depending on the factors present during their formation.
For example, zones of Holocene (with materials from recent glaciations) and
Pleistocene (older glaciers) deposits have promoted the development of allophanic
and non-allophanic Andosols, respectively (Poulenard et al. 2003; Harden 2007).
Topography and vegetation are other important key factors involved in the forma-
tion of these soils: for instance, according to Zehetner et al. (2003), additions of
high amounts of organic matter from the páramo vegetation in combination with its
slow decomposition rate promoted the presence of allophanic materials over 3200 m
asl in the Ecuadorian Andes. However, as altitude decreases, those allophanic mate-
rials transformed into other types of colloids – like halloysites – thus modifying the
properties of Andosols to such a degree that they transformed into Entisols or
Inceptisols.
Human activities commonly performed in Andean páramos – such as burning,
grazing, cultivation, and afforestation – also affect the variability of Andosols.
Although Andosols present a good physical fertility due to their high water reten-
tion capacity, good hydraulic conductivity, and low bulk density (Podwojewski and
Poulenard 2006; Buytaert et al. 2011), they seriously limit agriculture development
owing to their strong capacity to fix phosphate ions, their low pH, and their high
aluminum content (Saigusa and Matsuyama 1998; IUSS Working Group WRB
2014).
Among the many microorganisms able to promote plant growth and develop-
ment, arbuscular mycorrhizal fungi (AMF) deserve a special place. Estimated to be
present in up to 80% of all terrestrial plant species, arbuscular mycorrhiza is the
most frequent type of symbiosis established between plants and members of the
Glomeromycota phylum (Wang and Qiu 2006; Brundrett 2009). More than 225 spe-
cies in this phylum have been described; however, it is likely that this number does
not reflect the actual richness of AMF (Kivlin et al. 2011).
AMF colonize cortical cells of the roots of vascular plants where they develop
their characteristic arbuscular structures that allow nutrient exchange between the
host plant and the fungus. By extending the volume of soil that can be explored by
AMF hyphae (soil scouting), these fungi help their host plants to reach distant nutri-
ents – primarily phosphorus but also sulfur, nitrogen, and micronutrients – and to
uptake and transfer water (Parniske 2008; Baum et al. 2015). Additionally, they can
alleviate plant biotic and abiotic stresses (Gianinazzi et al. 2010; Singh et al. 2011b)
and also act as biocontrollers of a wide range of pathogens, mostly fungi but also
bacteria and nematodes (Harrier and Watson 2004; Whipps 2004; Jung et al. 2012;
Schouteden et al. 2015). In addition to the direct benefits provided to their host
plants, AMF also favors plant growth and development by improving soil structure
and aggregation through the formation of an intricate network of hyphae and an
abundant secretion of exopolysaccharides (Wright and Upadhyaya 1998) (Fig. 10.2).
10.5 P
GPM Prospection in the Tropical Andean Region
Context
Many of the abovementioned types of PGPM have been tested both in greenhouse
and field trials and proven to be very efficient at promoting plant growth (Glick
2012). Consequently, numerous commercial PGPM-based biofertilizers have been
developed and are currently marketed worldwide. However, their effectiveness in
the soil-plant system is uncertain, particularly under field conditions. Still, they
remain a very attractive alternative to chemical fertilizers and pesticides, particu-
larly in less developed regions (Sharma et al. 2016; Verma et al. 2015a; Meena et al.
2013b, 2016e; Masood and Bano 2016).
Often, commercial products formulated from exogenous PGPM result in poor
performances when applied to different crops. This is frequently due to a low sur-
vival rate of exogenous microbial strains when applied to soils different than those
from which these strains were isolated. Among the reasons explaining this phenom-
enon, we can mention (1) the different edaphic characteristics or climatic conditions
these microorganisms encounter, (2) a fierce competition by better-adapted indige-
nous microorganisms during plant colonization, and/or (3) predation by other
microorganisms (Bashan 1998). Therefore, in order to guarantee an adequate per-
formance of bioinoculants, isolation and screening of indigenous PGPM strains are
more than justified (Shrivastava et al. 2016; Velazquez et al. 2016; Meena et al.
2015c; Bahadur et al. 2016a).
216 L.A. Yarzábal et al.
Common to both:
Nutrient mobilization and acquisition
Resistance against pathogenic fungi
Drought tolerance
Cold tolerance (antifreezing)
Plant defense induction (Induced Systemic Resistance)
Competitionwithpathogens
Altered rhizosphereinte ractions
Fig. 10.2 Prominent plant-beneficial traits exhibited by rhizospheric soil microorganisms. Some
of the most important functions performed by these microorganisms are also included
Traditionally, this prospection has been performed by first analyzing the soil
characteristics where the plants will be grown and taking into consideration their
specific requirements, to identify then the required bacterial traits that might be
beneficial to these particular conditions. Following such a scheme, Andean environ-
ments have been increasingly prospected during recent years to isolate and charac-
terize potential indigenous PGPM in order to develop new biotechnological
products, well fitted to increase crop yields under the particular environmental con-
ditions of the Andes. Since most of these microorganisms are frequently associated
with plant roots, several research groups have focused on studying the microbiology
of the rhizosphere of the most important food crops in the Central Andes. The cases
of potato and quinoa are reviewed in the following pages considering their relevance
in terms of global markets, food security, and tradition (Sindhu et al. 2016; Meena
et al. 2014b, 2015d; Singh et al. 2015, 2016; Meena et al. 2013c).
10 Microbial Diversity of Tropical Andean Soils and Low-Input Sustainable… 217
With a worldwide annual production of ~385 Mt per year (FAO Stat 2014), potato
(Solanum tuberosum) is the fourth most important food crop after maize, wheat, and
rice and is considered critical for food security in an increasingly populated world
(Birch et al. 2012). Potato, which originated in the highlands of Peru and Bolivia,
has been cultivated locally by farming communities for centuries (Spooner et al.
2005) and is nowadays the main staple crop in the Andes Mountains.
Potato plants are often grown at high altitudes under harsh climatic conditions
and in fields where nutrients are often unavailable due to high soil acidity, which
may explain why their average production yields are so low. Because it requires
large inputs of expensive chemical fertilizers, the productivity of potatoes in small
farms is severely constrained. Additionally, fungal diseases that affect potato yield
and tuber quality (such as late blight, black scurf, and dry rot) are still common in
the Andes (Finckh et al. 2006). The aforementioned reasons explain why research-
ers have devoted great efforts to characterize the microbes colonizing potato root
system in order to search for potential growth promoters and biocontrollers.
The International Potato Center – founded in 1971 with the aim of delivering
sustainable solutions to the pressing world problems of hunger, poverty, and the
degradation of natural resources (www.cipotato.org) – implemented in 2005 a strat-
egy directed toward exploring the use of rhizobacteria to improve potato production
in a sustainable way. The final goal of this project was to develop biofertilizers for
small-scale farmers in the Central Andean Highlands. Initial studies were performed
with commercial products based on either a Peruvian strain of Azotobacter sp.
(Azotolum from Peru) or a Bacillus subtilis strain from Germany (FZB24). In a field
trial performed at 4000 m asl, biofertilization of three Peruvian potato varieties –
Yungay, Putis, and Queccorani –increased tuber yields by up to 25% compared to
the control plants (Oswald et al. 2007). In a second field trial, performed at 3200 m
asl, a total of 23 potato varieties – mainly Peruvian potato landraces – were planted
and biofertilized. The results were not as clear-cut as the previous ones, and differ-
ent responses to PGPB were observed depending on the potato variety (Oswald
et al. 2007).
Following these encouraging results, bacteria colonizing the rhizosphere of pota-
toes were subsequently isolated from rhizospheric soils sampled in different prov-
inces of the Peruvian Andes, tested in vitro for their PGP abilities, and inoculated to
potatoes grown in aeroponic systems (Calvo-Vélez et al. 2008; Calvo et al. 2009).
The results obtained showed significant increases in tuber numbers per plant, rang-
ing from 40% to 100%, depending on the potato cultivar (Oswald and Calvo 2009).
In a follow-up of this work, Oswald et al. (2010) isolated bacteria from the rhizo-
spheric soil of potatoes cultivated in Huancavelica and Puno, at locations situated
between 3900 and 4200 m asl in the Central Andean Highlands of Peru.
On the basis of the PGP activities recorded in vitro, 56 isolates – belonging to the
genera Bacillus and Azotobacter and actinomycetes – were selected for a pot experi-
ment conducted under greenhouse conditions and including seven commercial
potato varieties. Inoculation of plants with more than 80% of these strains increased
218 L.A. Yarzábal et al.
total plant and tuber weights between 50% and 200%. Using some of these strains,
a field trial was conducted in Puno (3820 m asl), where the authors recorded higher
tuber yields in inoculated potato plants than those obtained in the uninoculated
controls.
The characterization of bacteria colonizing the rhizosphere of potato was also
performed by other research groups in Peru. For example, Calvo and Zuñiga (2010)
and Calvo et al. (2010) studied the PGP abilities of several Bacillus sp. strains, for-
merly isolated in the highland Peruvian regions of Huancavelica and Puno – Titicaca
Lake – at 3900–4200 m asl by Calvo-Vélez et al. (2008). Besides exhibiting several
PGP characteristics in vitro, the isolates inhibited growth of R. solani and F. solani.
Taken together, the results showed the potential of these strains to develop PGP
inoculants.
More recently, Ghyselinck et al. (2013) investigated the bacterial communities
colonizing potatoes grown at different altitudes in the Central Andean Highlands of
Peru and Bolivia. Fifty eight strains out of 585 isolates exhibited antagonism against
R. solani and P. infestans. Additionally, 14 isolates (belonging to the Pseudomonas
koreensis subgroup, the P. fluorescens subgroup and the Bacillus genus) protected
potato plantlets challenged with R. solani. On the other side, 12 strains significantly
increased plant growth and development when tested in vitro. These results clearly
indicated that some of these isolates might be used to develop new biocontrol
products.
Field trials to investigate the effects of PGPM on potato productivity were also
conducted in the Andean region of Bolivia, particularly at Lope Mendoza (3440 m
asl, Carrasco Province), Tiraque highlands (3453 m asl, Tiraque Province), La Villa
(2650 m asl, Cochabamba Province), and Colomi (3200 m asl, Chapare Province)
(Franco et al. 2011). In these trials, native isolates of B. amyloliquefaciens and
B. subtilis, as well as a commercial B. subtilis strain (FZB 24WG), were tested as
single microbial treatments on the native potato cultivar Waycha (S. tuberosum
sub-sp. andigena Hawkes) with the participation of local farmers. Treatments com-
bining native B. subtilis strains with organic amendments resulted in superior and
uniform plant emergence, higher plant vigor and development, improved plant
health, and higher yield of tubers. Also, a suppressive effect on the incidence of
R. solani in tubers was observed. The commercial B. subtilis strain did not perform
well in regard to plant growth and yield.
Aside from the beneficial effects of PGP bacteria on potatoes, it has been repeat-
edly shown that AMF can also promote potato production (Hijri 2016). These ben-
efits do not only rely on the ability of AMF to increase nutrient uptake (mainly P)
by plant roots, but also has to do with an enhanced water uptake, a greater disease
resistance, and a significant alleviation of cultural and environmental stresses
(Berruti et al. 2015). However, these benefits depend on several factors among
which we can mention the potato genotype (Vosatka and Gryndler 2000), the AM
fungal species or strain (Davies et al. 2005a), and the soil P levels (Herrera-Peraza
et al. 2011).
Whatever the case, positive responses of S. tuberosum were recorded by
McArthur and Knowles (1993) after inoculation with AMF, in experiments
10 Microbial Diversity of Tropical Andean Soils and Low-Input Sustainable… 219
10.6 O
mic Technologies and Agriculture Development
in the Andean Region
Until now, prospection of the microbial communities colonizing Andean crops has
relied on culture-based techniques. Consequently, the major flaw in these studies is
that the great majority of microbes – the unculturable ones – remain to be identified
and their functions revealed. Therefore, our understanding of these communities
10 Microbial Diversity of Tropical Andean Soils and Low-Input Sustainable… 221
and their changes over time, plant species and age, soil properties, and/or manage-
ment is at its infancy.
Phenotypic and genotypic approaches are currently available to characterize soil
microbial community structures. Metagenomics—defined as the direct genetic anal-
ysis of genomes contained within an environmental sample—provides access not
only to the microbial diversity of a given community (the so-called phylogenetic
surveys) but, more importantly, also to the functional gene composition of microbial
communities (Thomas et al. 2012). For example, in a recent study based on pyrose-
quencing, Barnett et al. (2015) studied the variability of the potato root-associated
bacterial community with regard to different locations and potato clones. From a
total of 26 potato clones collected at three sites, 123 bacterial operational taxonomic
units (OTUs) were correlated with tuber yield and/or tuber nutrient content, a major-
ity of them belonging to the order Rhizobiales. Interestingly, 23 of these bacterial
OTUs were present in at least 50% of samples at each site and, therefore, were
considered as part of a core bacteriome likely involved in supporting potato crop
health.
With a similar objective in mind, two studies were conducted in three different
Andean countries (Peru, Bolivia, and Ecuador), at different altitudes (from 2658 to
4075 m asl), and at three potato plant developmental stages (emergence, flowering,
and senescence) (Senés-Guerrero et al. 2014; Senés-Guerrero and Schüssler 2016).
In both studies, the AMF communities associated with potatoes were described,
although the methods used differed: the first report relied on soil DNA extraction,
PCR amplification, cloning and sequencing of selected molecular markers (SSU,
LSU, and ITS region), while the second report was based on metagenomic analyses
of LSU rRNA gene PCR amplicons by pyrosequencing methods (454-GS FLX+).
According to the results of the first study, potato plants were colonized by diverse
species from 8 of the 11 Glomeromycota families, a much higher AMF diversity
than those reported for potato in other ecosystems (Senés-Guerrero et al. 2014).
Besides, the composition of the AMF community was affected by potato plant
developmental stages, and most of the AMF species detected in potato roots at the
emergence stage did not persist during later stages. Contrary to what is characteris-
tic in other alpine environments, the highest AMF diversity occurred at the highest
altitude (4075 m asl) in both soil and root samples. The study also highlighted the
importance of three OTUs – namely, Funneliformis mosseae, an unknown
Claroideoglomus sp., and Rhizophagus irregularis, easily cultivable generalists
which colonize early plant developmental stages, as promising candidates for the
development of biofertilizers for sustainable potato agriculture.
The second work cited analyzed in depth, for the first time, the composition of
the AMF community colonizing Andean potatoes (Senés-Guerrero and Schüssler
2016). In this study, the SSU-ITS-LSU rRNA gene region used was defined as an
extended DNA barcode which allowed resolving closely related AMF species
(Stockinger et al. 2010; Schoch et al. 2012). Following this methodology, the results
showed that almost 92% of the individual root systems studied contained between 1
and 25 AMF species. Additionally, OTU analyses clearly revealed that the AMF
community structure varied according to altitude, plant stage, or plant variety.
222 L.A. Yarzábal et al.
10.7 D
oes Biotechnology (and Bioinoculants) Represent
a Real Alternative for Small Andean Farmers?
microorganisms to increase the productivity and health of potato plants grown under
high- and low-input cropping systems. These assays did not only include the pilot-
scale production of AMF and PGPR inoculums, but were followed by participatory
field trials conducted with local farming communities and NGOs. The farmers
themselves were asked to evaluate the success or failure of these trials and selected
the most accepted technologies in each community. As can be deduced from these
results, the VALORAM experience, as a whole, seems to confirm that when a bio-
logical approach is combined with suitable plant genotypes, appropriate land man-
agement, and inoculation with beneficial microorganisms, then a valuable
opportunity results for the sustainable intensification of potato-based farming sys-
tems in the Andean region (VALORAM Final Report 2015). Furthermore, prelimi-
nary cost-benefit analyses indicate that farmers may reduce their expenses to almost
50% – for example, by reducing their dependence on agrochemicals.
References
Adesemoye O, Torbert HA, Kloepper JW (2009) Plant growth-promoting rhizobacteria allow
reduced application rates of chemical fertilizers. Microb Ecol 58:921–929
Ahemad M, Kibret M (2014) Mechanisms and applications of plant growth promoting rhizobacte-
ria: current perspective. J King Saudi Univ Sci 26:1–20
Ahmad M, Nadeem SM, Naveed M, Zahir ZA (2016) Potassium-solubilizing bacteria and their
application in agriculture. In: Meena VS, Maurya BR, Verma JP, Meena RS (eds) Potassium
solubilizing microorganisms for sustainable agriculture. Springer, New Delhi, pp 293–313.
doi:10.1007/978-81-322-2776-2_21
Aubron C, Cochet H, Brunschwig G, Moulin C-H (2009) Labor and its productivity in Andean
dairy farming systems: a comparative approach. Hum Ecol 37:407–419
Bahadur I, Meena VS, Kumar S (2014) Importance and application of potassic biofertilizer in
Indian agriculture. Int Res J Biol Sci 3:80–85
Bahadur I, Maurya BR, Meena VS, Saha M, Kumar A, Aeron A (2016a) Mineral release dynamics
of tricalcium phosphate and waste muscovite by mineral-solubilizing rhizobacteria isolated
from indo-gangetic plain of India. Geomicrobiol J. doi:10.1080/01490451.2016.1219431
Bahadur I, Maurya BR, Kumar A, Meena VS, Raghuwanshi R (2016b) Towards the soil sustain-
ability and potassium-solubilizing microorganisms. In: Meena VS, Maurya BR, Verma JP,
Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture. Springer,
New Delhi, pp 225–266. doi:10.1007/978-81-322-2776-2_18
Barnett BA, Holm DG, Koym JW et al (2015) Site and clone effects on the potato root-associated
core microbiome and its relationship to tuber yield and nutrients. Am J Potato Res 92:1–9
10 Microbial Diversity of Tropical Andean Soils and Low-Input Sustainable… 227
Dotaniya ML, Meena VD, Basak BB, Meena RS (2016) Potassium uptake by crops as well as
microorganisms. In: Meena VS, Maurya BR, Verma JP, Meena RS (eds) Potassium solu-
bilizing microorganisms for sustainable agriculture. Springer, New Delhi, pp 267–280.
doi:10.1007/978-81-322-2776-2_19
Eakin H, Lemos MC (2006) Adaptation and the state: Latin America and the challenge of capacity-
building under globalization. Glob Environ Chang 16:7–18
FAO Stat (2014) Solanum tuberosum annual production. Available at: http://faostat3.fao.org/
search/solanum%20tuberosum/E
Finckh MR, Schulte-Geldermann E, Bruns C (2006) Challenges to organic potato farming: disease
and nutrient management. Potato Res 49:27–42
Fonte SJ, Vanek SJ, Oyarzun P et al (2012) Pathways to agroecological intensification of soil fer-
tility management by smallholder farmers in the Andean highlands. Adv Agron 116:125–184
Food and Agriculture Organization of the United Nations (FAO) Report (2003) Organic agri-
culture: the challenge of sustaining food production while enhancing biodiversity. FAO
Corporate Document Repository. http://www.fao.org/3/contents/4bd9da73-3374-592e-a44f-
c087adf7a552/AD090E00.HTM. Accessed 27 Sept 2016
Food and Agriculture Organization of the United Nations (FAO) Report (2013) The state of food
and agriculture: food systems for better nutrition. FAO Corporate Document Repository.
http://www.fao.org/docrep/018/i3300e/i3300e00.htm. Accessed 27 Sept 2016
Franco J, Main G, Navia O, Ortuño N, Herbas J (2011) Improving productivity of traditional
Andean small farmers by bio-rational soil management: I. The potato case. Rev Lat Papa
16:270–290
Franco J, Main G, Urquieta E (2015) Valorización de la diversidad microbiológica andina a través
de la intensificación sostenible de sistemas agrícolas basados en el cultivo de papa (VALORAM)
(pp. 52–57). In: Fundación PROINPA. Informe Compendio 2011–2014. Cochabamba –
Bolivia. Available at: http://www.proinpa.org/publico/Informe_compendio_2011_2014/valori-
zacion%20de%20la%20diversidad%20microbiologica.pdf
Fuentes-Ramírez LE, Caballero-Mellado J (2006) Chapter 5. Bacterial biofertilizers. In: Siddiqui
ZA (ed) PGPR: biocontrol and biofertilization. Springer, Dordrecht, pp 143–172
Gardi C, Angelini M, Barceló S et al (2014) Atlas de suelos de América Latina y el Caribe.
ComisiónEuropea – Oficina de Publicaciones de la UniónEuropea
Ghyselinck J, Velivelli SLS, Heylen K et al (2013) Bioprospecting in potato fields in the central
Andean highlands: screening of rhizobacteria for plant growth-promoting properties. Syst Appl
Microbiol 36:116–127
Gianinazzi S, Gollotte A, Binet M-N et al (2010) Agroecology: the key role of arbuscular mycor-
rhizas in ecosystem services. Mycorrhiza 20:519–530
Glick BR (2012) Plant growth-promoting bacteria: mechanisms and applications. Scientifica,
Article ID 963401, 15 pages. http://dx.doi.org/10.6064/2012/963401
Haller A (2014) The “sowing of concrete”: Peri-urban smallholder perceptions of rural–urban land
change in the central Peruvian Andes. Land Use Policy 38:239–247
Harden C (2007) Two soil toposequences in a tropical glacial trough, southern Ecuador.
Zeitschriftfür Geomorphol 51:139–152
Harrier LA, Watson CA (2004) The potential role of arbuscular mycorrhizal (AM) fungi in the
bioprotection of plants against soil-borne pathogens in organic and/or other sustainable farm-
ing systems. Pest Manag Sci 60:149–157
Herrera-Peraza RA, Hamel C, Fernandez F, Ferrer RL, Furrazola E (2011) Soil strain compat-
ibility: the key to effective use of arbuscular mycorrhizal inoculants? Mycorrhiza 21:183–193
Herridge D, Peoples M, Boddey R (2008) Global inputs of biological nitrogen fixation in agricul-
tural systems. Plant Soil 311:1–18
Hijri M (2016) Analysis of a large dataset of mycorrhiza inoculation field trials on potato shows
highly significant increases in yield. Mycorrhiza 26:209–214
Hirose Y, Fujita T, Ishii T, Ueno N (2010) Antioxidative properties and flavonoid composition of
Chenopodium quinoa seeds cultivated in Japan. Food Chem 119:1300–1306
10 Microbial Diversity of Tropical Andean Soils and Low-Input Sustainable… 229
IUSS Working Group WRB (2014) World reference base for soil resources 2014. International soil
classification system for naming soils and creating legends for soil maps, World Soil Resources
Reports, vol 106. FAO, Rome
Jacobsen SE (2011) The situation for quinoa and its production in southern Bolivia: from eco-
nomic success to environmental disaster. J Agron Crop Sci 197:390–399
Jaiswal DK, Verma JP, Prakash S, Meena VS, Meena RS (2016) Potassium as an important plant
nutrient in sustainable agriculture: a state of the art. In: Meena VS, Maurya BR, Verma JP,
Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture. Springer,
New Delhi, pp 21–29. doi:10.1007/978-81-322-2776-2_2
Jat LK, Singh YV, Meena SK, Meena SK, Parihar M, Jatav HS, Meena RK, Meena VS (2015)
Does integrated nutrient management enhance agricultural productivity? J Pure Appl Microbiol
9(2):1211–1221
Jha Y, Subramanian RB (2016) Regulation of plant physiology and antioxidant enzymes for alle-
viating salinity stress by potassium-mobilizing bacteria. In: Meena VS, Maurya BR, Verma JP,
Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture. Springer,
New Delhi, pp 149–162. doi:10.1007/978-81-322-2776-2_11
Jodha NS, Shrestha S (1993) Sustainable and more productive mountain agriculture: problems and
prospects. In: Mountain environment and development –part 3 (Thematic Papers). ICIMOD,
Kathmandu, pp 1–65
Johnson SE, Loeppert RH (2006) Role of organic acids in phosphate mobilization from iron oxide.
Soil Sci Soc Am J 70:222–234
Jones DL (1998) Organic acids in the rhizosphere—a critical review. Plant Soil 205:25–44
Jung SC, Martinez-Medina A, Lopez-Raez JA, Pozo MJ (2012) Mycorrhiza-induced resistance
and priming of plant defenses. J Chem Ecol 38:651–664
Kivlin SN, Hawkes CV, Treseder KK (2011) Global diversity and distribution of arbuscular mycor-
rhizal fungi. Soil Biol Biochem 43:2294–2303
Kumar A, Bahadur I, Maurya BR, Raghuwanshi R, Meena VS, Singh DK, Dixit J (2015) Does
a plant growth-promoting rhizobacteria enhance agricultural sustainability? J Pure Appl
Microbiol 9:715–724
Kumar A, Meena R, Meena VS, Bisht JK, Pattanayak A (2016b) Towards the stress management
and environmental sustainability. J Clean Prod 137:821–822
Kumar A, Patel JS, Bahadur I, Meena VS (2016a) The molecular mechanisms of KSMs for
enhancement of crop production under organic farming. In: Meena VS, Maurya BR, Verma JP,
Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture. Springer,
New Delhi, pp 61–75. doi:10.1007/978-81-322-2776-2_5
Kumar A, Maurya BR, Raghuwanshi R, Meena VS, Islam MT (2017) Co-inoculation with
Enterobacter and Rhizobacteria on yield and nutrient uptake by wheat (Triticum aestivum L.)
in the alluvial soil under indo-gangetic plain of India. J Plant Growth Regul. doi:10.1007/
s00344-016-9663-5
Lozada C (2006) Overgrazing and range degradation in the Peruvian Andes. Rangel Arch 13:64–66
Lucy M, Reed E, Glick B (2004) Application of free living plant growth-promoting rhizobacteria.
Antonie Van Leeuwenhoek 86:1–25
Masood S, Bano A (2016) Mechanism of potassium solubilization in the agricultural soils by the
help of soil microorganisms. In: Meena VS, Maurya BR, Verma JP, Meena RS (eds) Potassium
solubilizing microorganisms for sustainable agriculture. Springer, New Delhi, pp 137–147.
doi:10.1007/978-81-322-2776-2_10
Maurya BR, Meena VS, Meena OP (2014) Influence of Inceptisol and Alfisol’s potassium solubi-
lizing bacteria (KSB) isolates on release of K from waste mica. Vegetos 27:181–187
McArthur DAJ, Knowles NR (1993) Influence of vesicular arbuscular mycorrhizal fungi on the
response of potato to phosphorus deficiency. Plant Physiol 101:147–160
Meena VS, Maurya BR, Bohra JS, Verma R, Meena MD (2013a) Effect of concentrate manure
and nutrient levels on enzymatic activities and microbial population under submerged rice in
alluvium soil of Varanasi. Crop Res 45(1,2 & 3):6–12
230 L.A. Yarzábal et al.
Meena OP, Maurya BR, Meena VS (2013b) Influence of K-solubilizing bacteria on release of
potassium from waste mica. Agric Sust Dev 1:53–56
Meena VS, Maurya BR, Verma R, Meena RS, Jatav GK, Meena SK, Meena SK (2013c) Soil
microbial population and selected enzyme activities as influenced by concentrate manure and
inorganic fertilizer in alluvium soil of Varanasi. The Bioscan 8(3):931–935
Meena VS, Maurya BR, Bahadur I (2014a) Potassium solubilization by bacterial strain in waste
mica. Bang J Bot 43:235–237
Meena VS, Maurya BR, Verma JP (2014b) Does a rhizospheric microorganism enhance K+ avail-
ability in agricultural soils? Microbiol Res 169:337–347
Meena VS, Maurya BR, Meena RS (2015a) Residual impact of wellgrow formulation and NPK on
growth and yield of wheat (Triticum aestivum L.) Bangladesh J Bot 44(1):143–146
Meena VS, Maurya BR, Verma JP, Aeron A, Kumar A, Kim K, Bajpai VK (2015b) Potassium
solubilizing rhizobacteria (KSR): isolation, identification, and K-release dynamics from waste
mica. Ecol Eng 81:340–347
Meena VS, Meena SK, Verma JP, Meena RS, Ghosh BN (2015c) The needs of nutrient use efficiency
for sustainable agriculture. J Clean Prod 102:562–563. doi:10.1016/j.jclepro.2015.04.044
Meena VS, Verma JP, Meena SK (2015d) Towards the current scenario of nutrient use efficiency in
crop species. J Clean Prod 102:556–557. doi:10.1016/j.jclepro.2015.04.030
Meena RS, Meena VS, Meena SK, Verma JP (2015e) The needs of healthy soils for a healthy
world. J Clean Prod 102:560–561
Meena RS, Meena VS, Meena SK, Verma JP (2015f) Towards the plant stress mitigate the agricul-
tural productivity: a book review. J Clean Prod 102:552–553
Meena VS, Bahadur I, Maurya BR, Kumar A, Meena RK, Meena SK, Verma JP (2016a) Potassium-
solubilizing microorganism in evergreen agriculture: an overview. In: Meena VS, Maurya BR,
Verma JP, Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture.
Springer, New Delhi, pp 1–20. doi:10.1007/978-81-322-2776-2_1
Meena RS, Bohra JS, Singh SP, Meena VS, Verma JP, Verma SK, Sihag SK (2016b) Towards the
prime response of manure to enhance nutrient use efficiency and soil sustainability a current
need: a book review. J Clean Prod 112(1):1258–1260
Meena VS, Meena SK, Bisht JK, Pattanayak A (2016c) Conservation agricultural practices in
sustainable food production. J Clean Prod 137:690–691
Meena SK, Rakshit A, Meena VS (2016d) Effect of seed bio-priming and N doses under varied
soil type on nitrogen use efficiency (NUE) of wheat (Triticum aestivum L.) under greenhouse
conditions. Biocatal Agric Biotechnol 6:68–75
Meena RK, Singh RK, Singh NP, Meena SK, Meena VS (2016e) Isolation of low temperature sur-
viving plant growth-promoting rhizobacteria (PGPR) from pea (Pisum sativum L.) and docu-
mentation of their plant growth promoting traits. Biocatalysis and Agricultural. Biotechnology
4:806–811
Moreno Díaz P (1988) Inoculación de micorrizas en papa (Solanumtuberosum) respuesta en el cre-
cimiento y nutrición de plantas inoculadas en invernadero y en campo. Rev Lat Papa 1:84–103
Mulligan M, Rubiano J, Hyman G et al. (2009) The Andes basin focal project. Final report to the
CGIAR challenge program on water and food. CPWF Project Report PN63
Mulligan M, Rubiano J, Hyman G et al (2010) The Andes basins: biophysical and developmental
diversity in a climate of change. Water Int 35:472–492
Nunes da Rocha U, van Overbeek L, van Elsas JD (2009) Exploration of hitherto-uncultured bac-
teria from the rhizosphere. FEMS Microbiol Ecol 69:313–328
Ochoa-Tocachi BF, Buytaert W, De Bièvre B et al (2016) Impacts of land use on the hydrological
response of tropical Andean catchments. Hydrol Process 30:4074–4089
Oliveira CA, Alves VMC, Marriel IE et al (2009) Phosphate solubilizing microorganisms isolated
from rhizosphere of maize cultivated in an oxisol of the Brazilian Cerrado Biome. Soil Biol
Biochem 41:1782–1787
Ortuño N, Navia O, Medrano A, Rojas K, Torrico L (2010) Desarrollo de bioinsumos: un aporte a
la soberanía alimentaria de Bolivia. Rev Agric 62:30–38
10 Microbial Diversity of Tropical Andean Soils and Low-Input Sustainable… 231
Ortuño N, Castillo JA, Claros M et al (2013) Enhancing the sustainability of quinoa production and
soil resilience by using bioproducts made with native microorganisms. Agronomy 3:732–746
Ortuño N, Claros M, Gutiérrez C et al (2014) Bacteria associated with the cultivation of quinoa in
the Bolivian Altiplano and their biotechnological potential. Rev Agric 53:53–61
Oswald A, Calvo P (2009) Using rhizobacteria to improve productivity of potato. 15th Triennial
ISTRC Proceedings. pp 29–33
Oswald A, Calvo P, Sanchez J, Zúñiga D (2007) Using plant growth promoting rhizobacteria to
improve potato production and plant health. Proceedings of the 16th International Symposium of
the International Scientific Centre for Fertilizers (CIEC) -Mineral Versus Organic Fertilization
Conflict or Synergism? Gent, Belgium. pp 401–409
Oswald A, Calvo Velez P, ZúñigaDávila D, Arcos Pineda J (2010) Evaluating soil rhizobacteria
for their ability to enhance plant growth and tuber yield in potato. Ann Appl Biol 157:259–271
Owen D, Williams AP, Griffith GW, Withers PJA (2014) Use of commercial bio-inoculants to
increase agricultural production through improved phosphorus acquisition. Appl Soil Ecol
86:41–54
Parewa HP, Yadav J, Rakshit A, Meena VS, Karthikeyan N (2014) Plant growth promoting rhizo-
bacteria enhance growth and nutrient uptake of crops. Agric Sustain Dev 2(2):101–116
Parniske M (2008) Arbuscular mycorrhiza: the mother of plant root endosymbioses. Nat Rev
Microbiol 6:763–775
Partap T (1999) Sustainable land management in marginal mountain areas of the Himalayan
region. Mt Res Dev 19:251–260
Pérez B, Gabriel J, Angulo A et al (2015) Efecto de los bioinsumos sobre la capacidad de respuesta
de cultivares nativos de papa (Solanumtuberosum L.) a sequía. Rev Lat Papa 19:40–58
Persello-Cartieaux F, Nussaume L, Robaglia C (2003) Tales from the underground: molecular
plant–rhizobacteria interactions. Plant Cell Environ 26:189–199
Pestalozzi H (2000) Sectoral fallow systems and the management of soil fertility: the rationality of
indigenous knowledge in the High Andes of Bolivia. Mt Res Dev 20:64–71
Piperno DR (2011) The origins of plant cultivation and domestication in the new world tropics:
patterns, processes, and new developments. CurrAnthropol 52:S453–S470
Podwojewski P, Poulenard J (2006) Paramos soils. In: Lal R (ed) Encyclopedia of soil, Science, vol
2. Taylor and Francis, New York, pp 1239–1242
Posada JM, Aide TM, Cavelier J (2000) Cattle and weedy shrubs as restoration tools of tropical
montane rainforest. Restor Ecol 8:370–379
Poulenard J, Podwojewski P (2006) Alpine soils. In: Lal R (ed) Encyclopedia of soil, Science, vol
2. Taylor and Francis, New York, pp 75–79
Poulenard J, Podwojewski P, Herbillon AJ (2003) Characteristics of non-allophanic Andisols with
hydric properties from the Ecuadorian páramos. Geoderma 117:267–281
Prakash S, Verma JP (2016) Global perspective of potash for fertilizer production. In: Meena VS,
Maurya BR, Verma JP, Meena RS (eds) Potassium solubilizing microorganisms for sustainable
agriculture. Springer, New Delhi, pp 327–331. doi:10.1007/978-81-322-2776-2_23
Priyadharsini P, Muthukumar T (2016) Interactions between arbuscular mycorrhizal fungi and
potassium-solubilizing microorganisms on agricultural productivity. In: Meena VS, Maurya
BR, Verma JP, Meena RS (eds) Potassium solubilizing microorganisms for sustainable agricul-
ture. Springer, New Delhi, pp 111–125. doi:10.1007/978-81-322-2776-2_8
PROINPA. BiotopCatalog. Retrieved at: http://www.proinpa.org/tic/index.php?option=com_conte
nt&view=article&id=122:catalogo-de-bioinsumos&catid=44&Itemid=153
Raghavendra MP, Nayaka NC, Nuthan BR (2016) Role of rhizosphere microflora in potas-
sium solubilization. In: Meena VS, Maurya BR, Verma JP, Meena RS (eds) Potassium
solubilizing microorganisms for sustainable agriculture. Springer, New Delhi, pp 43–59.
doi:10.1007/978-81-322-2776-2_4
Rajendhran J, Gunasekaran P (2008) Strategies for accessing soil metagenome for desired applica-
tions. Biotechnol Adv 26:576–590
232 L.A. Yarzábal et al.
Rawat J, Sanwal P, Saxena J (2016) Potassium and its role in sustainable agriculture. In: Meena
VS, Maurya BR, Verma JP, Meena RS (eds) Potassium solubilizing microorganisms for sus-
tainable agriculture. Springer, New Delhi, pp 235–253. doi:10.1007/978-81-322-2776-2_17
Richardson AE, Simpson RJ (2011) Soil microorganisms mediating phosphorus availability.
Update on microbial phosphorus. Plant Physiol 156:989–996
Rodriguez H, Fraga R (1999) Phosphate solubilizing bacteria and their role in plant growth promo-
tion. Biotechnol Adv 17:319–339
Rojas W, Valdivia R, Padulosi S et al (2009) From neglect to limelight: issues, methods and
approaches in enhancing sustainable conservation and use of Andean grains in Bolivia and
Peru. J Agric Rural Dev Trop Subtrop 92:87–117
Rosset PM, Altieri MA (1997) Agroecology versus input substitution: a fundamental contradiction
of sustainable agriculture. Soc Nat Resour 10:283–295
Ruiz KB, Biondi S, Oses R et al (2014) Quinoa biodiversity and sustainability for food security
under climate change. A review. Agron Sustain Dev 34:349–359
Saha M, Maurya BR, Bahadur I, Kumar A, Meena VS (2016a) Can potassium-solubilising bacteria
mitigate the potassium problems in India? In: Meena VS, Maurya BR, Verma JP, Meena RS
(eds) Potassium solubilizing microorganisms for sustainable agriculture. Springer, New Delhi,
pp 127–136. doi:10.1007/978-81-322-2776-2_9
Saha M, Maurya BR, Meena VS, Bahadur I, Kumar A (2016b) Identification and characterization
of potassium solubilizing bacteria (KSB) from Indo-Gangetic Plains of India. Biocatal Agric
Biotechnol 7:202–209
Saigusa M, Matsuyama N (1998) Distribution of Allophanic Andosols and non-allophanic
Andosols in Japan. Tohoku J Agric Res 48:75–83
Sandor JA, Eash NS (1995) Ancient agricultural soils in the Andes of southern Peru. Soil Sci Soc
Am J 59:170–179. doi:10.2136/sssaj1995.03615995005900010026x
Schmidt R, Cordovez V, de Boer W et al (2015) Volatile affairs in microbial interactions. ISME
J 9:2329–2335
Schoch C, Seifert K, Huhndorf S et al (2012) Nuclear ribosomal internal transcribed spacer (ITS)
region as a universal DNA barcode marker for fungi. Proc Natl Acad Sci U S A 109:6241–6246
Schouteden N, De Waele D, Panis B, Vos CM (2015) Arbuscular mycorrhizal fungi for the bio-
control of plant-parasitic nematodes: a review of the mechanisms involved. Front Microbiol
6:1280
Senés-Guerrero C, Schüssler A (2016) A conserved arbuscular mycorrhizal fungal core-species
community colonizes potato roots in the Andes. Fungal Divers 77:317–333
Senés-Guerrero C, Torres-Cortés G, Pfeiffer S et al (2014) Potato-associated arbuscular mycor-
rhizal fungal communities in the Peruvian Andes. Mycorrhiza 24:405–417
Sharma A, Shankhdhar D, Shankhdhar SC (2016) Potassium-solubilizing microorganisms:
mechanism and their role in potassium solubilization and uptake. In: Meena VS, Maurya BR,
Verma JP, Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture.
Springer, New Delhi, pp 203–219. doi:10.1007/978-81-322-2776-2_15
Shoji S, Dahlgren R, Nanzyo M (1993) Terminology, concepts and geographic distribution of vol-
canic ash soils. In: Shoji S, Nanzyo M, Dahlgren R (eds) Volcanic ash soils: genesis, properties
and utilization. Elsevier Science Publishers B.V, Amsterdam, pp 1–6
Shrivastava M, Srivastava PC, D’Souza SF (2016) KSM soil diversity and mineral solubilization,
in relation to crop production and molecular mechanism. In: Meena VS, Maurya BR, Verma JP,
Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture. Springer,
New Delhi, pp 221–234. doi:10.1007/978-81-322-2776-2_16
Sieverding E, Friedrichsen J, Suden W (1991) Vesicular-arbuscular mycorrhiza management in
tropical agrosystems. Deutsche GesellschaftfuerTechnischeZusammenarbeit GmbH, Eschborn
Sindhu SS, Parmar P, Phour M, Sehrawat A (2016) Potassium-solubilizing microorganisms
(KSMs) and its effect on plant growth improvement. In: Meena VS, Maurya BR, Verma JP,
Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture. Springer,
New Delhi, pp 171–185. doi:10.1007/978-81-322-2776-2_13
10 Microbial Diversity of Tropical Andean Soils and Low-Input Sustainable… 233
Singh LP, Gill SS, Tuteja N (2011a) Unraveling the role of fungal symbionts in plant abiotic stress
tolerance. Plant Signal Behav 6:175–191
Singh JS, Pandey VC, Singh DP (2011b) Efficient soil microorganisms: a new dimension for sus-
tainable agriculture and environmental development. Agric Ecosyst Environ 140(3e4):339e353
Singh NP, Singh RK, Meena VS, Meena RK (2015) Can we use maize (Zea mays) rhizobacteria as
plant growth promoter? Vegetos 28(1):86–99. doi:10.5958/2229-4473.2015.00012.9
Singh M, Dotaniya ML, Mishra A, Dotaniya CK, Regar KL, Lata M (2016) Role of biofertil-
izers in conservation agriculture. In: Bisht JK, Meena VS, Mishra PK, Pattanayak A (eds)
Conservation agriculture: an approach to combat climate change in Indian Himalaya. Springer,
Singapore, pp 113–134. doi:10.1007/978-981-10-2558-7_4
Sinha SK (1997) Global change scenario: current and future with reference to land cover changes
and sustainable agriculture- south and south-east Asian context. Curr Sci 72:846–854
Spooner DM, McLean K, Ramsay G et al (2005) A single domestication for potato based on
multilocus amplified fragment length polymorphism genotyping. Proc Natl Acad Sci USA
102:14694–14699
Stanish C (2007) Agricultural intensification in the Titicaca Basin. In: Thurston TL, Fisher CT
(eds) Seeking a richer harvest. The archaeology of subsistence intensification, innovation, and
change. Springer, New York, pp 125–139
Stewart E (2012) Growing unculturable bacteria. J Bacteriol 194:4151–4160
Stockinger H, Krüger M, Schüssler A (2010) DNA barcoding of arbuscular mycorrhizal fungi.
New Phytol 187:461–474
Teotia P, Kumar V, Kumar M, Shrivastava N, Varma A (2016) Rhizosphere microbes: potassium
solubilization and crop productivity-present and future aspects. In: Meena VS, Maurya BR,
Verma JP, Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture.
Springer, New Delhi, pp 315–325. doi:10.1007/978-81-322-2776-2_22
Thomas T, Gilbert J, Meyer F (2012) Metagenomics – a guide from sampling to data analysis.
Microb Inform Exp 2:3. http://doi.org/10.1186/2042-5783-2-3
Trivedi P, Pandey A, Palni LMS (2012) Bacterial inoculants for field applications under moun-
tain ecosystems: present initiatives and future prospects. In: Maheshwari DK (ed) Bacteria in
agrobiology: plant probiotics. Springer, Berlin, pp 15–44. doi:10.1007/978-3-642-27515-9_2
Turner BL, Frossard E, Oberson A (2006) Enhancing phosphorus availability in low-fertility soils.
In: Uphoff NT et al (eds) Biological approaches to sustainable soil systems. CRC Press, Boca
Raton, pp 191–205
VALORAM Report Summary (2015) Retrieved at http://cordis.europa.eu/result/rcn/164393_
en.html on 2016/09/05
Velazquez E, Silva LR, Ramírez-Bahena MH, Peix A (2016) Diversity of potassium-solubilizing
microorganisms and their interactions with plants. In: Meena VS, Maurya BR, Verma JP,
Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture. Springer,
New Delhi, pp 99–110. doi:10.1007/978-81-322-2776-2_7
Velivelli SLS, Kromann P, Lojan P et al (2014a) Identification of mVOCs from Andean rhizobacte-
ria and field evaluation of bacterial and mycorrhizal inoculants on growth of potato in its center
of origin. Microb Ecol 69:652–667
Velivelli SLS, Sessitsch A, Doyle Prestwich B (2014b) The role of microbial inoculants in inte-
grated crop management systems. Potato Res 57:291–309
Verma R, Maurya BR, Meena VS (2014) Integrated effect of bio-organics with chemical fertilizer
on growth, yield and quality of cabbage (Brassica oleracea var capitata). Indian J Agric Sci
84(8):914–919
Verma JP, Jaiswa DK, Meena VS, Meena RS (2015a) Current need of organic farming for enhanc-
ing sustainable agriculture. J Clean Prod 102:545–547
Verma JP, Jaiswal DK, Meena VS, Kumar A, Meena RS (2015b) Issues and challenges about sus-
tainable agriculture production for management of natural resources to sustain soil fertility and
health. J Clean Prod 107:793–794
234 L.A. Yarzábal et al.
S.P. Shanthakumar
Abstract
Global food produce is contributed largely by agriculture and agro-based indus-
tries which is crucial for food security. But importance given to the organic crop
protection for sustainable farming and land use is trivial across the countries.
Recent upgradation in agro-farming systems by techno-crafts for higher yield
and crop protection should also place microbes as an important entity in pest
management. Development of new biocontrol agents from the microbial com-
munity is being reported which should gain lead in organic product development.
Earlier microbial pesticide was found to be a slow action pest control or preven-
tion tool in biological control approach. Many target-specific pest control agents
like entomopathogenic bacteria and virus were found to be swift in action when
compared with entomopathogenic fungi; henceforth, product developers and
biocontrol specialists have reclaimed the hope on biological pest control because
they could reduce pest population instantly and ecofriendly. In present articles
focus on various examples, process and related information on entomopatho-
genic bacteria for development of biocontrol products against major chickpea
crop pest Helicoverpa armigera. Special reference is given to chickpea crop
since India is the largest producer of chickpea facing the highest crop loss due to
pod borer.
Keywords
Insect pathology • Bacteria • Arthropods • Rhizosphere • Non-target organisms
11.1 Introduction
Global food production of all types stood at ~5.2 billion tons, representing some
~2.7 billion tons of edible dry matter, of which legumes and oil seeds contribute
gross tonnage of ~165 Mt. with maximum of ~39 Mt. of dry protein (Borlaug 2003).
The Indian agriculture is at the crossroad; our population may reach ~1750 million
by 2050. Per capita cropland will then be 0.089 ha, and per capita fresh water supply
will be ~1190 m3/year. Food grain production must be doubled, and the area under
irrigation should go up from the current ~60–114 M ha by 2050 (Swaminathan
2010).
Major investment of the farmers in agriculture is on pesticides for control of pest.
Achieving sustainable pest control and finding equilibrium levels often take many
years, and the result is not the instant pest ‘extermination’ that is wished for by
many farmers (Neuenschwander 2004). Low crop productivity is related to abiotic
and biotic factors, viz. damage caused by insect pests, which are realized as a major
biotic constraint to sustainable crop production (Sharma and Yadav 2001). Crop loss
due to insect pests is estimated to ~14% of the total agricultural production. The
global monetary loss due to insect pests accounts to US$ ~90 billion in rice, wheat,
maize, potato, cotton, soybean, barley and coffee (Andersen and Cohen 2000). India
has the distinction of world’s largest producer of pulses, but the average productiv-
ity of ~614 kg/ha is very low because of biotic stress in terms of insect infestation.
Among all the insect pests, the lepidopteran insect, H. armigera, poses a serious
threat to the pulses especially to chickpea and pigeon pea (Rabindra et al. 2005;
Behere et al. 2013). Not all pests are amenable to classical biological control; many
more biocontrol agents should be identified and explored. Parasitoids, predators,
pathogens and nematodes have all been used for control of pestiferous arthropods
(Figs. 11.1 and 11.2).
11.2 C
hickpea Production: Global, National and State
Prospects
The chickpea (Cicer arietinum L.) is the third most important food legume grown in
the world. In India, chickpea is an important pulse crop in terms of both area and
production. India is the largest producer of chickpea in the world sharing the range
between ~65% and 66% (FAOSTAT 2008) of the total area of ~12 M ha and produc-
tion of ~9 Mt., respectively. As per FAO report, Mexico has registered highest pro-
ductivity (~1809 kg/ha) followed by Canada (~1580 kg/ha) and Australia (~1268 kg/
ha). In India, chickpea cultivation is being done in ~6 M ha with the production of
~4 Mt. of the grains during 2002–2003. During 2008–2010, chickpea production
reached to ~7 Mt., and it was ~71% production at global scenario (FAOSTAT 2010).
Madhya Pradesh, Uttar Pradesh, Rajasthan, Maharashtra, Gujarat, Andhra Pradesh,
Tamil Nadu and Karnataka are the major chickpea-producing states in India.
Coimbatore, Erode, Perambalur and Dindigul are the highest chickpea-producing
districts in Tamil Nadu (INDIASTAT 2012).
11 Entomopathogenic Bacteria and Biorationals in Chickpea Organic Crop Protection 237
Perambalur Dindigul
Erode 6% 9%
7% Salem
3%
Dharmapuri
1%
Coimbatore
74%
In the case of Dindigul district, more than 500 acres of cultivable land is being
used for chickpea cultivation, and an average of 203 tonnes of chickpea production
per year is the statistical projections by national database (INDIASTAT 2012).
Insects are the most widely distributed and successful animal class in the world. The
indispensable reasons for this phenomenon are rapid reproduction, short life cycle
and a robust immune system (Kranthi and Kranthi 2004; Gayatri Priya et al. 2012;
Behere et al. 2013). The crop production losses have shown an increasing trend over
the years. In 1983, the losses due to insect pests were estimated worth Rs. 6000
crores (Krishnamurthy Rao and KSRK 1983) which increased to Rs. 20,000 crores
in 1993 (Jayaraj 1993) and to Rs. 29,000 crores in 1996 (Dhaliwal and Arora 2010).
Birthal and Sharma (2004) have reported that H. armigera alone causes an annual
loss of about Rs. 1000 crores in India. The American bollworm, H. armigera, is a
polyphagous pest of worldwide occurrence inflicting annual crop damage in India
worth US $1 billion (Subramanian and Mohankumar 2006). Global crop losses due
to H. armigera are estimated to be over US $ 2 billion annually, of which 50% loss
occurs in India, causing widespread misery and frequent crop failures (Subramaniam
Gopalakrishnan et al. 2011).
In India, this insect occurs as a major pest in many economically important
crops, including cotton, pigeon pea, chickpea, tomato, lady’s finger and black gram
(Ge et al. 2003; Subramanian and Mohankumar 2006; Nguyen et al. 2012; Guo
et al. 2012). Severe outbreaks were recorded on soybean and mustard in Madhya
Pradesh (Singh 2005), pigeon pea in Karnataka and Andhra Pradesh (Singh 1994),
black cumin and strawberries in Himachal Pradesh (Singh 1999), finger millet and
sweet orange in Andhra Pradesh (Reddy and Reddy 1999) and chickpea, linseed and
mustard in Orissa (Patnaik and Lenka 2000).
It has also been observed that H. armigera attacks few new host plants such as
rose and gherkin (Malarvannan 2004). During the off season, H. armigera is
observed to survive on few weeds like Abutilon indicum, Achyranthes aspera,
Cleome monophylla, Solanum viarum, Datura metel, Acanthospermum hispidum
and Gynandropsis gynandra (Rao et al. 1991).
In Tamil Nadu, the data collected on cotton from 1985–1986 to 1987–1988
revealed that H. armigera reached the economic threshold level of 10% damage to
fruiting parts. The districts of Coimbatore, Erode and Salem had a higher incidence
in 1987–1988, and to some extent, a similar trend was noticed in the earlier 2 years
(Jayaraj et al. 1990). Intensive field surveys on H. armigera incidence in southern
districts (Aruppukottai, T.Kallupatti and Thirumangalam) of Tamil Nadu, India, in
different rainfed cotton cropping systems (cotton alone and cotton intercropped
with cow pea, black gram, green gram, sun flower and sorghum systems) recorded
maximum H. armigera under pure cotton (Saminathan et al. 2003; Meena et al.
2013a; Bahadur et al. 2014; Maurya et al. 2014; Jat et al. 2015; Kumar et al. 2016b).
11 Entomopathogenic Bacteria and Biorationals in Chickpea Organic Crop Protection 239
In the Indian conditions, most of the farmers use insecticides as a dominant tool
among the various options for the management of the insect pest. However, the
indiscriminate use of insecticides has resulted in resistance development of H. armi-
gera to almost all groups of insecticides. In India, resistance was first reported in
cotton-growing belt of Andhra Pradesh in 1987 (Reddy 1988). Low to moderate
levels of resistance have been reported in Maharashtra; moderate to high in Tamil
Nadu, Uttar Pradesh, Gujarat and Punjab; and high in Andhra Pradesh and Madhya
Pradesh (Kranthi et al. 2006). Excessive use of insecticides has resulted in an
increase in severity of sucking pest which was considered as minor pests in addition
to environmental pollution. In order to avoid the development of insecticide resis-
tance, it is essential to curtail the indiscriminate use of the insecticide (Nimbalkar
et al. 2009; Kumar et al. 2015, 2016a; Ahmad et al. 2016; Meena et al. 2016a;
Parewa et al. 2014; Jaiswal et al. 2016; Jha and Subramanian 2016).
The pest is primarily controlled by biological and chemical means, yet the
insect’s wide-ranging host preference, high population density throughout the
breeding season and tendency to develop resistance to insecticides hamper attempts
to overcome the damage it causes (Gunning and Moores 2002; Landers 2007).
However, increased use of chemical pesticides has posed serious health hazards and
widespread ecological damage (Rejaul Hassan and Karim 2005). Though chemical
insecticides have contributed largely in the management of pest, it has resulted in a
number of ecological and health hazards due to the accumulation of undesirable
residues in the food crops besides promoting insecticide resistance of pests with
negative impact on non-target species (Arti and Sujoita 2009).
India produces more than 200 pesticides and formulations, the largest in Asia
and twelfth in the world. The agricultural sector consumes ~67% of the pesticides
produced, with two-third alone taken by crops like cotton, paddy, vegetables and a
fruit; the largest usage is recorded in Andhra Pradesh followed by Tamil Nadu. The
insecticides chlorpyriphos, phosalone and malathion were also highly superior in
reducing the pest incidence (Biradar et al. 2001). Bioassay against H. armigera on
pigeon pea revealed that fenvalerate and monocrotophos are more toxic, followed
by endosulfan and dimethoate (Sharma and Yadav 2001). On chickpea, the efficacy
of acephate alone and in combination with other synthetic pyrethroids was superior
against H. armigera. Similarly, deltamethrin resulted in lowest fruit damage due to
H. armigera on tomato followed by cypermethrin and acephate (Mehta et al. 2001;
Prakash and Verma 2016; Meena et al. 2015a, f, 2016b; Priyadharsini and
Muthukumar 2016; Kumar et al. 2017; Dotaniya et al. 2016).
A troublesome aspect of H. armigera is their ability to develop resistance to
insecticides. The pest has build-up resistance to commonly used pesticides metho-
myl, chlorpyriphos, quinalphos, acephate, fenvalerate, endosulfan and monocroto-
phos and synthetic pyrethroids up to 20- to 60-folds depending on the location
(Singh et al. 2002). Resistance to DDT, organophosphorus insecticides and syn-
thetic pyrethroids has bedevilled chemical pest control application and, in some
240 S.P. Shanthakumar
cases, has forced abandonment of cropping due to inability to control the pest
(David 1997).
Farmers mainly use inorganic pesticides against insect pests (Malik et al. 2002).
The outbreak of H. armigera has been attributed to the development of insecticide
resistance and the use of broad spectrum insecticides, which are known to have a
detrimental effect on populations of its natural enemies and nutritional and biocli-
matic factors in host plants (Naseri et al. 2009). Therefore, alternative measures
have been adopted, and considerable attention has been directed towards ecofriendly
insecticides in the past few years (Arti and Sujoita 2009). In Indian agriculture, the
IPM programme has commenced during the last decade, and one of the important
steps in biocontrol is using beneficial organisms for the control of insect pest (Prasad
et al. 2007; Raghavendra et al. 2016; Zahedi 2016; Meena et al. 2015b; Rawat et al.
2016; Das and Pradhan 2016; Dominguez-Nunez et al. 2016).
India has been bestowed with varied diversity, rich in beneficial bacterial, viral,
fungal and entomopathogenic nematodes. This, apparently, offers a greater scope
for microbial control making it more feasible and cost-effective to the resource-poor
farmers. Microbial pesticide would help to manage insecticide-resistant pest popu-
lation of H. armigera, prevent the rise of secondary pest problems and fit well in
agriculture (Zhou et al. 2013; Pawar and Borikar 2005). Microbial pathogens are
distinguished from commensal microorganisms by their ability to enter, proliferate,
persist and disseminate from host sites that are inaccessible to commensal organ-
isms (Finlay and Falkow 1997).
Almost half (~49%) of the total introductions of traditional fungi for biocontrol
of insect pests were conducted (Hajek et al. 2007). The most commonly introduced
species were Metarhizium anisopliae and Beauveria bassiana in India (Hajek and
Delalibera 2010). However, ~700 products of different microbial are currently
available worldwide. Thirty-eight fungal formulations based on Metarhizium,
Beauveria and Lecanicillium and ~45 baculovirus-based formulations for the con-
trol of Helicoverpa and Spodoptera are available (Vasantharaj and Ramamurthy
2008). Fungal pathogens particularly Beauveria bassiana (Saxena and Ahmad
1999; Uma Devi et al. 2008), Metarhizium anisopliae (Gopalakrishnan and
Narayanan 1989; Khaderkhan et al. 1993; Shanthakumar et al. 2009), Lecanicillium
lecanii and Nomuraea rileyi (Lingappa et al. 2005; Shanthakumar et al. 2010b) have
been widely used as insecticides globally.
Next important microbial agent is the entomopathogenic virus. The most preva-
lent viruses in the environment, three families (Baculoviridae, Polydnaviridae,
Ascoviridae), are specific for insects and related arthropods. The most widely
11 Entomopathogenic Bacteria and Biorationals in Chickpea Organic Crop Protection 241
exploited virus groups for biocontrol of insects pests are the baculoviruses (Evans
2000; Monobrullah and Nagata 2000). Some of the viral organisms pathogenic to H.
armigera are nuclear polyhedrosis virus (NPV), cytoplasmic polyhedrosis virus
(CPV) and granulosis virus (GV).
The average mortality of H. armigera due to the above pathogens was ~21% and
could reach ~57% in an epidemic year (Fan et al. 1999). Application of different
doses of Splt MNPV showed dose-related mortality on final instars of Spodoptera
litura (Mohammad and Uma 2008). Entomopathogenic viruses are generally con-
sidered to be environmentally safe and do not leave toxic residues (Morris 1980;
Monobrullah 2003). However, widespread use of viruses for pest management has
been hampered by their relatively high cost, slow activity and susceptibility to UV
degradation (Moscardi 1999).
Followed by entomopathogenic virus, entomopathogenic nematodes are other
important biocontrol agents for agricultural insect pest management. It is well
known that entomopathogenic nematodes, which are classified into Steinernematidae
and Heterorhabditidae families, have great potential as biological control agents in
plant protection (Klein 1990). Their activity against different pest insects is already
well studied (Ebssa 2005; Laznik et al. 2008; Forst et al. 1997). Endemic nematode
was tested successively against various insect pests at the laboratory and field
(Ehlers 1995; Woodring and Kaya 1988 and Legaspi et al. 2000). Virus and nema-
todes even though perform well in controlling insect but when it comes to product
development at large scale for growing demand, both the group have some limita-
tions as they need living host system which is not required in case of bacterial ento-
mopathogens (Yasin et al. 2016; Meena et al. 2015e, 2016c, d; Saha et al. 2016a, b;
Yadav and Sidhu 2016; Teotia et al. 2016; Bahadur et al. 2016b).
Thus bacterial entomopathogens are versatile and can be produced in large scale
in fementers with limited resource which is economical and time saving in terms of
mass production. Naturally available soil bacteria from plant rhizosphere can be
chosen for a simple ecological compatibility reason to make the product effective in
field condition after augmentation. Many studies on soil bacterial insecticides were
carried out and reported against a wide range of pests as discussed in the following
chapters.
11.7 R
ole of Soil and Rhizosphere Entomopathogenic
Bacteria
Rhizobacteria have been studied as plant growth promoters for agricultural produc-
tion as well as biocontrol agents against both diseases and pests (Kloepper and
Beauchamp 1992). A few bacterial isolates with pesticidal activity against
Lepidoptera, Coleoptera and Diptera (Shah and Goettel 1999) are commercially
available. Notably the soil-dwelling insect pathogen, B. thuringiensis, is suitable for
use in biological control, and certain strains have been developed as commercial
bioinsecticides (Clara et al. 2004).
242 S.P. Shanthakumar
11.8 M
olecular Characterization and Identification
of Entomopathogenic Bacteria
Comparison and analysis of the 16S rRNA gene has been extensively used as a
diagnostic method to determine phylogenetic relationships of bacterial isolates.
Moreover, this gene is characterized by having areas of secondary structure which
have also proven useful for diagnostic purposes. The size of this gene is approxi-
mately 1.6 kb and is composed of regions with different levels of variability thus
providing valuable information for differentiation of taxa (Stackebrandt and Goebel
1994). The study of the genetic diversity among bacterial species and strains has
also had an impact in assessing relatedness among species and understanding their
diversity (Boemare and Tailliez 2009). Bacteria are characterized by having single
chromosome within the nucleoid. The size of this chromosome varies among spe-
cies. For example, the bacterial genomes of B. thuringiensis (the widely studied
insect pathogen), B. cereus and B. anthracis are around 5.4 Mb (Carlson and Kolsto
1993 and Ivanova et al. 2003). Other species such as B. subtilis and B. licheniformis
have a smaller chromosome (4.2 Mb) (Rey et al. 2004).
Entomopathogenic nematode symbiotic bacteria Photorhabdus luminescens
have a genome size of 5.7 Mb (Duchaud et al. 2003). Many bacteria also contain
extrachromosomal elements (plasmids), which are smaller double-stranded DNA
(dsDNA) molecules that replicate independent of the chromosomal DNA. Genes
located in bacterial plasmids usually code for proteins that determine specific phe-
notypes but do not code for products needed for bacterial survival and growth.
Bacterial genes are organized in operons or cassettes that consist of a promoter, a
series of genes and a transcription terminator (Verma et al. 2014, 2015b; Meena
et al. 2014a, 2016e; Bahadur et al. 2016a; Masood and Bano 2016).
The microbial entomopathogenic agents like bacteria and fungi infest the insect
primarily, and then after colonization, they secret toxins which curtails the normal
metabolism and decease. Entomopathogenic fungi like Beauveria sp. produce sev-
eral metabolites, including bassianin, beauvericin, bassianolide, beauveriolide,
bassiacridin, oosporein and tenellin (Vining et al. 1962; Strasser et al. 2000;
Quesada–Moraga and Vey 2004). Beauvericin is toxic to H. zea, Culex pipiens,
Aedes aegypti and Calliphora erythrocephala rupturing the midgut epithelium and
dissolving the ribosomes (Zizka and Weiser 1993; Grove 1985; Champlin and Grula
1979). Metarhizium sp. produces destruxins and cytochalasins (Roberts 1981); P.
tenuipes produces tenuipesine (Kikuchi et al. 2004); Akanthomyces gracilis pro-
duces akanthomycin (Wagenar et al. 2002); Cordyceps pseudomilitaris produces
coryanhidrides (Isaka et al. 2000); Tolypocladium spp. produce efrapeptins
(Krasnoff and Gupta 1992); Aschersonia aleyrodis produces destruxins and
homodestruxins; and Aschersonia tubulata produces dustanin (Krasnoff et al. 1996).
244 S.P. Shanthakumar
Different types of Cry proteins isolated from B. thuringiensis could directly bind
to peritrophic membrane and disintegrate the columnar cells and microvilli and
cause imbalance in gut pH and kill the lepidopteran insects (Garczynski and Siegel
2007). Oral injection with P. entomophila bacterial cells and monalysin toxin iso-
lated from P. entomophila could cause significant mortality in Drosophila melano-
gaster (Takayuki Kuraishi et al. 2011). Novel toxin complexes (Tc) A and (Tc) B
from bacterium Yersinia entomophaga against Plutella xylostella were reported by
Landsberg et al. (2011).
Davangere and Ranebennuru) and Tamil Nadu (Coimbatore and Dindigul) and were
found susceptible to crystal protein toxins (Jalali et al. 2004).
Secondary metabolite produced by bacteria for their insecticidal activity was not
much explored, and only few studies were taken up in the recent past. Detailed
analysis of liquid cultures of the entomopathogenic bacteria (Brachmann et al.
2006; Lang et al. 2008) showed that these bacteria are in fact very important second-
ary metabolite producers that can produce several structurally diverse compounds.
Therefore entomopathogenic bacteria can be regarded as a novel source of potential
biocontrol products which have been studied only recently. Eleftherianos et al.
(2007) show that Photorhabdus species produced a small-molecule (E)-1,3-
dihydroxy-2-(isopropyl)-5-(2-phenylethenyl)benzene (ST) that acted as an inhibitor
of phenoloxidase (PO) in the insect host Manduca sexta.
The entomopathogenic bacteria Xenorhabdus and Photorhabdus produce phage-
derived bacteriocins (Thlaer et al. 1995), colicin E3-type killer proteins (Singh and
Banerjee 2008) and insect toxin complexes (Ffrench-Constant and Waterfield 2006)
as well as several other secondary metabolites (Piel 2004). Insecticidal compounds
from bacteria described in the literature represent mostly small molecules like ben-
zylideneacetone (Ji et al. 2004), iodinine (Fodor et al. 2004), phenethylamines and
indole derivatives (Li et al. 1995; McInerney et al. 1991a), and comparatively less
number of complex compounds like the xenorhabdins and xenorxides (Li et al.
1998) and xenocoumacins (XCNs) (McInerney et al. 1991b) was reported.
Despite their simple structure, different biological activities have been described
for these compounds, and their biosynthesis genes have not been identified yet
except for the XCNs (Helge and Bode 2009). Additionally, it should be mentioned
that other entomopathogenic bacteria reported have not been analysed for second-
ary metabolites and/or genes encoding the corresponding biosynthesis genes but are
members of well-known insecticidal genera like Serratia entomophila and Erwinia
carotovora (Grimont et al. 1988; Vallet-Gely et al. 2008). Entomopathogenic activ-
ity of E. carotovora has been reported only under lab conditions and has to be
proven for its insecticidal activity under field conditions.
were tested against sheath blight and leaf folder in rice. The application of talc for-
mulation through seed, root, soil and foliar spray significantly reduced the sheath
blight and leaf folder incidence both under greenhouse and field conditions. Several
Pseudomonas strains have been found to control many fungal, bacterial, viral and
insect pests at laboratory level (Ramamoorthy et al. 2001). The laboratory biocon-
trol efficacy and PGPR activity of fluorescent pseudomonads were further increased
by mixing two or more strains of Pseudomonas species (Garica and Sarmiento
2000; Nandakumar et al. 2001; Nakkeeran et al. 2004) or in combination with other
bacterial or fungal antagonists (Duffy et al. 1996) or mixed with chitin or other
substances (Benhamou et al. 1998). Even though many reports on the entomopatho-
genic bacteria have been recorded, only limited studies were made on its efficacy at
field level which should be given importance in future studies for sustainable organic
crop protection (Shanthakumar et al. 2010a; Sharma et al. 2016; Verma et al. 2015a;
Meena et al. 2013b, c; Singh et al. 2015).
nematodes may affect some natural enemies. These effects are usually limited to
some natural enemies and are minor. Many studies have been published on compat-
ibility of entomopathogens and natural enemies; however, reports on greenhouse
and field evaluations are limited (Ravensberg 2011; Shrivastava et al. 2016;
Velazquez et al. 2016; Meena et al. 2014b, 2015c, d; Sindhu et al. 2016; Singh et al.
2016).
If the product developed does not pass through these safety parameters, it will
have deleterious effect on habitat management which is directly linked to the distur-
bances in diversity of natural enemies and the quality of the environment in which
they occur. As a result of frequent and intense disturbance regimes, many agricul-
tural systems are recognized as particularly difficult environments for natural ene-
mies (Landis and Marino 1999). This is significant for annual monocultural cropping
systems where the rates of establishment of imported natural enemies, and their
success in controlling the target pest are lower than in more stable cropping systems
(Hall et al. 1980).
Beneficial organisms are generally considered to be more susceptible to chemi-
cal insecticides than their hosts. Croft (1990) studied interactions between benefi-
cial insects and pesticides intensively in field crops, orchards, vineyards and
glasshouses. Most microbial control agents are generally considered to be host spe-
cific and pose a lower risk than associated with many chemical pesticides (Goettel
and Roberts 1992). Exposure of honey bees to high inoculum densities of B. bassi-
ana resulted in very low mortality that was not different from the control suggesting
that B. bassiana can be applied for pest control in fields where honey bees are used
for pollination (Al mazrawi 2007).
Earlier report on non-infectivity of HaNPV to Bombyx mori was confirmed by
Sanjay et al. (1991), where they found that B. mori fed on HaNPV-infected leaves
did not affect larval weight, larval duration, cocooning percentage, weight of
cocoons and percentage adult emergence of silkworm, thus reported to be non-
infective. Neem extract did not affect the parasitization of Trichogramma pretiosum
and T. principium (Klemm and Schmutterer 1993) as same as oils from neem, citro-
nella and lemon grass were not repellent to Coccinella undecimpunctata (Matter
et al. 1993) and hence can be used as biorationals in combination with bacterial
insect pathogens.
Microbial contrivances shows selective sweep in insect pest population without any
negative impact on environment and crop production. Need for laboratory and field
evaluation on the active microbes is felt necessary for product development to vali-
date their ecological impact precisely other than target pest control. This review
highlights the bright scope for bacterial entomopathogens both in action potential
and low-cost production over the entomopathogenic fungus, virus and nematodes.
Moreover, the secondary metabolite produced by the entomopathogens should be
given importance rather than protein toxins for future product development as the
248 S.P. Shanthakumar
Acknowledgements I thank Dr. Vasantharaj David for helpful comments on an earlier finding.
The author is also indebted to Prof. M. S. Swaminathan for the facilities provided to work on bio-
logical control.
References
Ahmad M, Nadeem SM, Naveed M, Zahir ZA (2016) Potassium-solubilizing bacteria and their
application in agriculture. In: Meena VS, Maurya BR, Verma JP, Meena RS (eds) Potassium
solubilizing microorganisms for sustainable agriculture. Springer, New Delhi, pp 293–313.
doi:10.1007/978-81-322-2776-2_21
Al mazrawi MS (2007) Impact of the entomopathogenic fungus, Beauveria bassiana on the honey-
bees, Apis mellifera (Hymenoptera: Apidae). World J Agri Sci 3(1):7–11
Andersen-Per Pinstrup, Marc J Cohen (2000) Modern biotechnology for food and agriculture:
risks and opportunities for the poor, Agr. Biotechnol Poor 159–169
Arti P, Sujoita P (2009) Evaluation of the morphological abnormalities in the 4th instar larva of
Helicoverpa armigera (Hub.) on application of leaf extract of Lantana camara (L.) World
J Zool 4(4):253–255
Ayyathurai V, Duraisamy L, Ambalavanan S, Ramasamy S (2009) Plant growth promoting rhizo-
bacteria of cotton affecting the developmental stages of Helicoverpa armigera. J Plant Protect
Res 49:3
Bahadur I, Meena VS, Kumar S (2014) Importance and application of potassic biofertilizer in
Indian agriculture. Int Res J Biol Sci 3:80–85
Bahadur I, Maurya BR, Kumar A, Meena VS, Raghuwanshi R (2016a) Towards the soil sustain-
ability and potassium-solubilizing microorganisms. In: Meena VS, Maurya BR, Verma JP,
Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture. Springer,
New Delhi, pp 225–266. doi:10.1007/978-81-322-2776-2_18
Bahadur I, Maurya BR, Meena VS, Saha M, Kumar A, Aeron A (2016b) Mineral release dynamics
of tricalcium phosphate and waste muscovite by mineral-solubilizing rhizobacteria isolated
from indo-gangetic plain of India. Geomicrobiol J. doi:10.1080/01490451.2016.1219431
Barloy F, Lecadet MM, Delecluse A (1998) Distribution of clostridial cry-like genes among
Bacillus thuringiensis and Clostridium strains. Curr Microbiol 36:232–237
Beat R, Pechy-Tarr M, Ryffel F, Hoegger P, Obrist C, Rindlisbacher A, Keel C, Maurhofer M (2013)
Oral insecticidal activity of plant-associated pseudomonads. Environ Microbiol 15:751–763
Behere GT, Tay WT, Russell DA, Kranthi KR, Batterham P (2013) Population genetic structure
of the cotton bollworm Helicoverpa armigera (Hubner) (Lepidoptera: Noctuidae) in India as
inferred from EPIC-PCR DNA markers. PLoS One 8:1
Bell E, Mullet JE (1993) Characterization of an Arabidopsis lipoxygenase gene responsive to
methyl jasmonate and wounding. Plant Physiol 103:1133–1137
Benhamou N, Kloepper JW, Tuzun S (1998) Induction of resistance against fusarium wilt of
tomato by combination of chitosan with an endophytic strain: ultrastructure and cytochemistry
of the host response. Planta 204:153–168
Biradar AP, Balikai RA, Teggelli RG (2001) Chemical control of pod borer, Helicoverpa armigera
(Hübner) on pigeon pea. Karnataka J Agr Sci 14:500–502
Birthal PS, Sharma OP (2004) Integrated pest management in Indian agriculture. Proceedings 11.
National centre for agricultural economics and policy research. Library Avenue, New Delhi
110 012
11 Entomopathogenic Bacteria and Biorationals in Chickpea Organic Crop Protection 249
Boemare N, Tailliez P (2009) In: Stock SP et al (eds) Insect pathogens: molecular approaches and
techniques. CAB International. Chapter 2, Wallingford, pp 31–45
Bong CFJ, Sikorowski PP (1991) Effects of cytoplasmic polyhedrosis virus and bacterial contami-
nation on growth and development of corn earworm, Heliothis zea (Boddie). J Invertebr Pathol
57:406–412
Borlaug NE (2003) Feeding a world of 10 million people, The TVA/IFDC legacy, Third lecture
published in Travis. P. Hignett memorial lecture series.
Brachmann AO, Forst S, Furgani GM, Fodor A, Bode HB (2006) Xenofuranones A and B: phenyl-
pyruvate dimmers from Xenorhabdus szentirmaii. J Nat Prod 69:1830–1832
Carlson CR, Kolsto AB (1993) A complete physical map of a Bacillus thuringiensis chromosome.
J Bacteriol 175:1053–1060
Champlin FR, Grula EA (1979) Noninvolvement of beauvericin in the entomopathogenicity of
Beauveria bassiana. Appl Environ Microbiol 37:1122–1125
Clara M, Porcar M, Lopez A, Escudero IRD, Perez-Llarena FJ, Caballero P (2004) Characterization
of a Bacillus thuringiensis strain with a broad spectrum of activity against lepidopteran insects.
Entomol Exp Appl 111:71–77
Croft BA (1990) Arthropod biological control agents and pesticides. Wiley, New York. 723pp
Daborn PJ, Waterfield N, Blight MA, Ffrench-Constant RH (2001) Measuring virulence factor
expression by the pathogenic bacterium Photorhabdus luminescens in culture and during insect
infection. J Bacteriol 183:5834–5839
Das I, Pradhan M (2016) Potassium-solubilizing microorganisms and their role in enhancing
soil fertility and health. In: Meena VS, Maurya BR, Verma JP, Meena RS (eds) Potassium
solubilizing microorganisms for sustainable agriculture. Springer, New Delhi, pp 281–291.
doi:10.1007/978-81-322-2776-2_20
David G (1997) Genetics of Heliothis and Helicoverpa resistance to chemical insecticides and to
Bacillus thuringiensis. Pest Sci 51:251–258
Dhaliwal GS, Arora R (2010) Principles of insect pest management. National Agricultural
Technology Information Centre, Ludhiana
Dominguez-Nunez JA, Benito B, Berrocal-Lobo M, Albanesi A (2016) Mycorrhizal fungi: role
in the solubilization of potassium. In: Meena VS, Maurya BR, Verma JP, Meena RS (eds)
Potassium solubilizing microorganisms for sustainable agriculture. Springer, New Delhi,
pp 77–98. doi:10.1007/978-81-322-2776-2_6
Dotaniya ML, Meena VD, Basak BB, Meena RS (2016) Potassium uptake by crops as well as
microorganisms. In: Meena VS, Maurya BR, Verma JP, Meena RS (eds) Potassium solu-
bilizing microorganisms for sustainable agriculture. Springer, New Delhi, pp 267–280.
doi:10.1007/978-81-322-2776-2_19
Duchaud E, Rusniok C, Frangeul L, Buchrieser C, Givaudan A, Taourit S, Bocs S, Boursaux-
Eude C, Chandler M, Charles JF, Dassa E, Derose R, Derzelle S, Freyssinet G, Gaudriault
S, Medigue C, Lanois A, Powell K, Siguier P, Vincent R, Wingate V, Zouine M, Glaser P,
Boemare N, Danchin A, Kunst F (2003) The genome sequence of the entomopathogenic bac-
terium Photorhabdus luminescens. Nat Biotechnol 21:1307–1313
Duffy BK, Simon A, Weller DM (1996) Combination of Trichoderma koningii with fluorescent
pseudomonads for control of take-all on wheat. Phytopathology 86:188–194
Ebssa L (2005) Efficacy of entomopathogenic nematodes for the control of the western flower
thrips Frankliniella occidentalis. Ph. D. Thesis, Hannover University: 141 pp
Ehlers RU (1995) Introduction of non-endemic nematodes for biological control: Scientific and
regulatory policy issues. COST & OECD workshop. Bruhnskoppel-Seminar hotel Malente,
Germany. 17 p
Eleftherianos I, Boundy S, Susan A J, Aslam S, Marshall JW, Cox RJ, Simpson TJ, Clarke DJ,
ffrench-Constant RH, Reynolds S (2007) An antibiotic produced by an insect-pathogenic
bacterium suppresses host defenses through phenoloxidase inhibition. Proc Natl Acad Sci
104(7):2419–2424
Evans HF (2000) Viruses. In: Lacey LA, Kaya HK (eds) Field manual of techniques in invertebrate
pathology. Kluwer Academic Publishers, Dordrecht, pp 179–208
250 S.P. Shanthakumar
Fan X, Feg SL, Fu YQ, Wang HM (1999) Field studies on the occurrence and epidemiology of
entomopathogenic diseases infecting Helicoverpa armigera in Hebei. Chinese J Biol Control
15:27–30
FAO (2008) FAOSTAT, Food and Agriculture Organisation of United Nations, Rome (http://fao-
stat3.fao.org/home/index.html#SEARCH_DATA)
FAO (2010) FAOSTAT, Food and Agriculture Organisation of United Nations, Rome (http://fao-
stat3.fao.org/home/index.html#SEARCH_DATA)
Ffrench-Constant R, Waterfield N (2006) An ABC guide to the bacterial toxin complexes 183.
ADV NU
Finlay BB, Falkow S (1997) Common themes in microbial pathogenicity revisited. Microbiol Mol
Biol Rev 61:136–169
Fodor A, Forst S, Haynes L, Hevesi M, Hogan J, Klein MG, Mathe-Fodor A, Stackebrndt, E.,
Szentirmai A, Sztaricskai F (2004) New perspectives of Xenorhabdus antibiotics research. In:
Ehlers RU, Enkerli J, Glazer I, Lopez-Ferber M, Tkaczuk C (eds) IOBC/WPRS bulletin: insect
pathogens insect parasitic nematodes. IOBC/WPRS; 2008, pp 157–164
Forst S, Dowds B, Boemare NE, Stackebrandt E (1997) Xenorhabdus spp. and Photorhabdus spp.:
bugs that kill bugs. Annu Rev Microbiol 51:47–72
Garczynski SF, Siegel JP (2007). In: Lacey LA, Kaya HK (eds) Field manual of techniques in
invertebrate pathology. Springer, Dordrecht, pp 175–197. doi: 10.1007/978-1-4020-5933-9
Garica OA, Sarmiento M (2000) A note on the viability of Azospirillum brasilense in turf used as
carrier in inoculated grass seeds. Cuba J Agric Sci 34:343–345
Gayatri Priya N, Ojha A, Kajla MK, Raj A, Rajagopal R (2012) Host plant induced variation in gut
bacteria of Helicoverpa armigera. PLoS One 7(1)
Ge F, Liu XH, Ding YQ, Wang XZ, Zhao YF (2003) Life-table of Helicoverpa armigera in
Northern China and characters of population development in Southern and Northern China.
Chin J Appl Ecol 14(2):241–245
Goettel MS, Roberts DW (1992) Mass production, formulation and field application of entomo-
pathogenic fungi. In: Lomer CJ, Prior C (eds) Biological control of locusts and grasshoppers.
CAB International, Wallingford, pp 230–238
Gopalakrishnan C, Narayanan K (1989) Occurrence of two entomofungal pathogen Metarhizium
anisopliae (Metschnikoff) Sorokin var. minor Tulloch and Nomuraea rileyi in Heliothis armi-
gera. Curr Sci 57:167–168
Grimont PAD, Jackson TA, Ageron E, Noonan MJ (1988) Serratia entomophila sp. nov. associated
with amber disease in the New Zealand grass grub Costelytra zealandica. Int J Syst Bacteriol
38:1–6
Gunning RV, Moores GD (2002) Chlorfenapyr resistance in Helicoverpa armigera in Australia.
In: Pests and diseases 2002. Proceedings of the British Crop Protection Council Conference.
British Crop Protection Council, United Kingdom. pp 793–798
Guo H, Sun Y, Ren Q, Zhu-Salzman K, Kang L (2012) Elevated CO2 reduces the resistance and
tolerance of tomato plants to Helicoverpa armigera by suppressing the JA signaling pathway.
PLoS One 7(7)
Gurr G, Wratten SD (2000) Biological control: measures of success. Kluwer Academic, Dordrecht
Gurr G, Wratten SD, Barbosa P (2000) Success in conservation biological control of arthropods.
In: Gurr GW, Wratten SD (eds) Biological control: measures of success. Kluwer Academic,
Dordrecht, pp 105–132
Hajek AE, Butler L (1999) Predicting the host range of entomopathogenic fungi. In: Follett
PA, Duan JJ (eds) Nontarget effects of biological control. Kluwer Acad. Publ, Dordrecht,
pp 263–276
Hajek AE, Delalibera I (2010) Fungal pathogens as classical biological control agents against
arthropods. BioControl 55:147–158
Hajek and Goettel (2007) Field manual of techniques in invertebrate pathology. In: Lacey LA and
Kaya HK (eds), pp 815–833
Hajek AE, McManus ML, Delalibera I (2007) A review of introductions of pathogens and nema-
todes for classical biological control of insects and mites. Biol Control 41:1–13
11 Entomopathogenic Bacteria and Biorationals in Chickpea Organic Crop Protection 251
Hall RW, Ehler LE, Bisabri-Ershadi B (1980) Rates of success in classical biological control of
arthropods. Bull Entomol Soc Am 26:111–114
Helge B, Bode (2009) Entomopathogenic bacteria as a source of secondary metabolites. Curr Opin
Chem Biol 13:224–230
Hokkanen HMT, Hajek AE (2003) Environmental impacts of microbial insecticides. Kluwer
Academic Publishers, Dordrecht
INDIASTAT (2012) Indian statistical department data base. (http://www.indiastat.com/agricul-
ture/2/agriculturalarealanduse/152/areaundercropsinindia19501951to20102011/448934/stats.
aspx)
Isaka M, Tanticharoen M, Thentaranonth Y (2000) Cordyanhydrides A and B. Two unique anhy-
drides from the insect pathogenic fungus Cordyceps pseudomilitaris BCC 1620. Tetrahedron
Lett 41:1657–1660
Ivanova N, Sorokin A, Anderson I, Galleron N, Candelon B, Kapatral V, Bhattacharyya A, Reznik
G, Mikhailova N, Lapidus A, Chu MM, Goltsman E, Larsen N, D’Souza M, Walunas T,
Grechkin Y, Pusch G, Haselkorn R, Fonstein M, Ehrlich SD, Overbeek R, Kyrpides N (2003)
Genome sequence of Bacillus cereus and comparative analysis with Bacillus anthracis. Nature
423:87–91
Jaiswal DK, Verma JP, Prakash S, Meena VS, Meena RS (2016) Potassium as an important plant
nutrient in sustainable agriculture: a state of the art. In: Meena VS, Maurya BR, Verma JP,
Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture. Springer,
New Delhi, pp 21–29. doi:10.1007/978-81-322-2776-2_2
Jalali SK, Mohan K, Singh SP, Manjunath TM, Lalitha Y (2004) Baseline- of the old-world boll-
worm, Helicoverpa armigera (Hubner) (Lepidoptera: Noctuidae susceptibility) populations
from India to Bacillus thuringiensis Cry1Ac insecticidal protein. Crop Prot 23:53–59
Jat LK, Singh YV, Meena SK, Meena SK, Parihar M, Jatav HS, Meena RK, Meena VS (2015)
Does integrated nutrient management enhance agricultural productivity? J Pure Appl Microbiol
9(2):1211–1221
Jayaraj J (1993) Biopesticides and integrated pest management for sustainable crop production. In:
Roy NK (ed) Agrochemicals in sustainable agriculture. APC Publications, New Delhi
Jayaraj S, Rangarajan AV, Gopalan M, Ramakrishnan C, Manoharan T, Thangaraju D (1990)
Biology and bionomics of Heliothis armigera Hübner and pest surveillance -a retrospect. In:
Jayaraj S, Uthamasamy S, Gopalan M, Rabindra RJ (eds) Heliothis management, Proc. Natl.
workshop. Tamil Nadu Agri. Univ., Coimbatore, pp 36–44
Jha Y, Subramanian RB (2016) Regulation of plant physiology and antioxidant enzymes for alle-
viating salinity stress by potassium-mobilizing bacteria. In: Meena VS, Maurya BR, Verma JP,
Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture. Springer,
New Delhi, pp 149–162. doi:10.1007/978-81-322-2776-2_11
Ji D, Yi Y, Kang GH, Choi YH, Kim P, Baek NI, Kim Y (2004) Identification of an antibacte-
rial compound, benzylideneacetone, from Xenorhabdus nematophila against major plant-
pathogenic bacteria. FEMS Microbiol Lett 239:241–248
Khaderkhan H, Jayaraj S, Gopalan M (1993) Muscardine fungi for the biological control of agro
forestry termite Odontotermes obesus (Rambur). Insect Sci Appl 14:529
Kikuchi H, Miyagawa Y, Nakamura K, Sahashi Y, Inatomi S, Oshima Y (2004) A novel carbon
skeletal trichothecene, tenuipesine A, isolated from an entomopathogenic fungus, Paecilomyces
tenuipes. Org Lett 6:4531–4533
Klein MG (1990) Efficacy against soil-inhabiting insect pests. In: Gaugler R, Kaya HK (eds)
Entomopathogenic nematode in biological control. CRC Press, Boca Raton, pp 195–214
Klemm U, Schmutterer H (1993) Effects of neem preparations on Plutella xylostella L. and its
natural enemies of the genus Trichogramma. Z Pflanzenkrankh Pflanzenschutz 100:113–128
Kloepper JW, Beauchamp CJ (1992) A review of issues related to measuring colonization of plant
roots by bacteria. Can J Microbiol 38:1219–1232
Kranthi KR, Kranthi NR (2004) Modeling adaptability of cotton boll worm Helicoverpa armigera
(Hubner) to Bt cotton in India. Curr Sci 87:1096–1107
252 S.P. Shanthakumar
Kranthi S, Kranthi KR, Bharose AA, Syed SN, Dhawad CS, Wadaskar RM, Behere GT, Patil EK
(2006) Cytochrome oxidase I sequence of Helicoverpa species in India – its utility as a molecu-
lar tool. Indian J Biotechnol 5:195–199
Krasnoff SB, Gupta S (1992) Efrapeptin production by Tolypocladium fungi (Deuteromycotina:
Hyphomycetes). Intra– and inter–specific variation. J Chem Ecol 18:1721–1741
Krasnoff SB, Gibson DM, Belofsky GN, Gloer KB, Gloer JB (1996) New destruxins from the
entomopathogenic fungus Aschersonia sp. J Nat Prod 59:485–489
Krishnamurthy Rao BH, Murthy, KSRK. (eds) (1983) Proceedings of national seminar on crop
losses due to insect pests. Indian J Ent. (Special issue), vols I–II. Hyderabad
Kumar A, Bahadur I, Maurya BR, Raghuwanshi R, Meena VS, Singh DK, Dixit J (2015) Does
a plant growth-promoting rhizobacteria enhance agricultural sustainability? J Pure Appl
Microbiol 9:715–724
Kumar A, Meena R, Meena VS, Bisht JK, Pattanayak A (2016a) Towards the stress management
and environmental sustainability. J Clean Prod 137:821–822
Kumar A, Patel JS, Bahadur I, Meena VS (2016b) The molecular mechanisms of KSMs for
enhancement of crop production under organic farming. In: Meena VS, Maurya BR, Verma JP,
Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture. Springer,
New Delhi, pp 61–75. doi:10.1007/978-81-322-2776-2_5
Kumar A, Maurya BR, Raghuwanshi R, Meena VS, Islam MT (2017) Co-inoculation with
Enterobacter and Rhizobacteria on yield and nutrient uptake by wheat (Triticum aestivum L.)
in the alluvial soil under indo-gangetic plain of India. J Plant Growth Regul. doi:10.1007/
s00344-016-9663-5
Kuraishi T, Binggeli O, Opota O, Buchon N, Lemaitre B (2011) Genetic evidence for a protective
role of the peritrophic matrix against intestinal bacterial infection in Drosophila melanogaster.
Proc Nat Acad Sci 108:15966–15971
Lacey LA (ed) (1997) Manual of techniques in insect pathology. Academic, London
Landers A (2007). Modern technology to improve spraying efficiency. Online:http://www.aben.
cornell.edu/extension/pestapp/publications/mod.tech.html.22/10/2007.
Landis DA, Marino PC (1999) Landscape structure and extra-field processes: impact on manage-
ment of pests and beneficials. In: Ruberson J (ed) Handbook of Pest management. Marcel
Dekker, New York, pp 79–104
Landsberg MJ, Sandra A, Rothnagel JR, Jason N, Busby Sean DG, Marshall Robert M, Simpsond
Shaun Lottc J, Hankamera B, Hurst MRH (2011) 3D structure of the Yersinia entomophaga
toxin complex and implications for insecticidal activity. Proc Natl Acad Sci 108:20544–20549
Lang G, Kalvelage T, Peters A, Wiese J, Imhoff JF (2008) Linear and cyclic peptides from the
entomopathogenic bacterium Xenorhabdus nematophilus. J Nat Prod 71:1074–1077
Laznik ZTT, Lakatos T, Trdan S (2008) Entomopathogenic nematode Steinernema feltiae
(Filipjev) (Rhabditida: Steinernematidae) recorded for the first time in Slovenia. Acta Agric
Slov 91:37–45
Legaspi JC, Legaspi BC, Saldana RR (2000) Evaluation of Steinernema riobravis (Nematoda:
Steinernematidae) against the Mexican rice borer (Lepidoptera: Pyralidae). J Entomol Sci
35(2):141–149
Li J, Chen G, Webster JM, Czyzewska E (1995) Antimicrobial metabolites from a bacterial sym-
biont. J Nat Prod 58:1081–1086
Li J, Hu K, Webster JM (1998) Antibiotics from Xenorhabdus spp. and Photorhabdus spp.
(Enterobacteriaceae). Chem Comp 34:1331–1339
Lingappa S, Saxena Hem, Devi Vimala PS, Rai R Ahmad and Sanjeev Gupta eds. Indian Society
of Pulses 2005. Role of biocontrol agents in management of Helicoverpa armigera (Hubner).
In: Recent advances in Helicoverpa armigera management (Hem Saxena Research and devel-
opment. IIPR, Kanpur, pp 159–184
Luttrell RG, Fitt GP, Ramalho FS, Sugonyaev ES (1994) Cotton pest management: Part 1.
A worldwide perspective. Annu Rev Entomol 39:517–526
Magurran AE (2004) Measuring biological diversity. Blackwell Publ, Malden
11 Entomopathogenic Bacteria and Biorationals in Chickpea Organic Crop Protection 253
Malarvannan S (2004) Studies on biocontrol of Helicoverpa armigera using traditional plants. Ph.
D Thesis. University of Madras, Chennai
Malik MF, Ali L, Anwar S (2002) Determination of installation heights for codling moths synthetic
pheromone traps in apple canopy. Asian J Plant Sci 1:226–227
Maria P-T, Bruck DJ, Maurhofer M, Fischer E, Vogne C, Henkels MD, Donahue MD, Grunder J,
Loper JE, Keel C (2008) Molecular analysis of a novel gene cluster encoding an insect toxin
in plant- associated strains of Pseudomonas fluorescens. Environ Microbiol 10(9):2368–2386
Masood S, Bano A (2016) Mechanism of potassium solubilization in the agricultural soils by the
help of soil microorganisms. In: Meena VS, Maurya BR, Verma JP, Meena RS (eds) Potassium
solubilizing microorganisms for sustainable agriculture. Springer, New Delhi, pp 137–147.
doi:10.1007/978-81-322-2776-2_10
Matter MM, Marei SS, Moawad SM, El Gengaihi S (1993) The reaction of Aphis gossypii and
its predator, Coccinella undecimpunctata to some plant extracts. Bull Fac Agric Univ Cairo
44:417–432
Maurya BR, Meena VS, Meena OP (2014) Influence of Inceptisol and Alfisol’s potassium solubi-
lizing bacteria (KSB) isolates on release of K from waste mica. Vegetos 27:181–187
McInerney BV, Gregson RP, Lacey MJ, Akhurst RJ, Lyons GR, Rhodes SH, Smith DR, Engelhardt
LM, White AH (1991a) Biologically active metabolites from Xenorhabdus spp., part 1.
Dithiolopyrrolone derivatives with antibiotic activity. J Nat Prod 54:774–784
McInerney TWC, Lacey MJ, Akhurst RJ, Gregson RP (1991b) Biologically active metabolites
from Xenorhabdus spp., part 2. Benzopyran-1-one derivatives with gastroprotective activity.
J Nat Prod 54:785–795
Meena VS, Maurya BR, Bohra JS, Verma R, Meena MD (2013a) Effect of concentrate manure
and nutrient levels on enzymatic activities and microbial population under submerged rice in
alluvium soil of Varanasi. Crop Res 45(1, 2 & 3):6–12
Meena OP, Maurya BR, Meena VS (2013b) Influence of K-solubilizing bacteria on release of
potassium from waste mica. Agric Sust Dev 1:53–56
Meena VS, Maurya BR, Verma R, Meena RS, Jatav GK, Meena SK, Meena SK (2013c) Soil
microbial population and selected enzyme activities as influenced by concentrate manure and
inorganic fertilizer in alluvium soil of Varanasi. The Bioscan 8(3):931–935
Meena VS, Maurya BR, Bahadur I (2014a) Potassium solubilization by bacterial strain in waste
mica. Bang J Bot 43:235–237
Meena VS, Maurya BR, Verma JP (2014b) Does a rhizospheric microorganism enhance K+ avail-
ability in agricultural soils? Microbiol Res 169:337–347
Meena VS, Maurya BR, Meena RS (2015a) Residual impact of wellgrow formulation and NPK on
growth and yield of wheat (Triticum aestivum L.) Bangladesh J Botany 44(1):143–146
Meena VS, Maurya BR, Verma JP, Aeron A, Kumar A, Kim K, Bajpai VK (2015b) Potassium
solubilizing rhizobacteria (KSR): isolation, identification, and K-release dynamics from waste
mica. Ecol Eng 81:340–347
Meena RS, Meena VS, Meena SK, Verma JP (2015c) The needs of healthy soils for a healthy
world. J Clean Prod 102:560–561
Meena RS, Meena VS, Meena SK, Verma JP (2015d) Towards the plant stress mitigate the agricul-
tural productivity: a book review. J Clean Prod 102:552–553
Meena VS, Meena SK, Verma JP, Meena RS, Ghosh BN (2015e) The needs of nutrient use efficiency
for sustainable agriculture. J Clean Prod 102:562–563. doi:10.1016/j.jclepro.2015.04.044
Meena VS, Verma JP, Meena SK (2015f) Towards the current scenario of nutrient use efficiency in
crop species. J Clean Prod 102:556–557. doi:10.1016/j.jclepro.2015.04.030
Meena VS, Bahadur I, Maurya BR, Kumar A, Meena RK, Meena SK, Verma JP (2016a) Potassium-
solubilizing microorganism in evergreen agriculture: an overview. In: Meena VS, Maurya BR,
Verma JP, Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture.
Springer, New Delhi, pp 1–20. doi:10.1007/978-81-322-2776-2_1
Meena RS, Bohra JS, Singh SP, Meena VS, Verma JP, Verma SK, Sihag SK (2016b) Towards the
prime response of manure to enhance nutrient use efficiency and soil sustainability a current
need: a book review. J Clean Prod 112(1):1258–1260
254 S.P. Shanthakumar
Meena VS, Meena SK, Bisht JK, Pattanayak A (2016c) Conservation agricultural practices in
sustainable food production. J Clean Prod 137:690–691
Meena SK, Rakshit A, Meena VS (2016d) Effect of seed bio-priming and N doses under varied
soil type on nitrogen use efficiency (NUE) of wheat (Triticum aestivum L.) under greenhouse
conditions. Biocatal Agric Biotechnol 6:68–75
Meena RK, Singh RK, Singh NP, Meena SK, Meena VS (2016e) Isolation of low temperature sur-
viving plant growth-promoting rhizobacteria (PGPR) from pea (Pisum sativum L.) and docu-
mentation of their plant growth promoting traits. Biocatal Agric Biotechnol 4:806–811
Mehta PK, Vaidya DN, Kashyap NP (2001) Management of tomato fruit borer, Helicoverpa armi-
gera (Hubner) using insecticides and biopesticides. Himachal J Agric Res 26:50–53
Mohammad M, Uma S (2008) Sub-lethal effects of SpltNPV infection on developmental of
Spodoptera litura (Lepidoptera: Noctuidae), bio-control. Sci Technol 4:431–437
Monobrullah M (2003) Optical brighteners – pathogenicity enhances of entomopathogenic viruses.
Curr Sci 84:640–645
Monobrullah MD, Nagata M (2000) Optimization of Spodoptera litura (Fab.) nucleo polyhedron
virus production in homologous host larvae. Insect Sci Appl 20:157–165
Morris ON (1980) Entomopathogenic viruses: strategies for use in forest insect pest management.
Can Entomol 112(6):573–584
Moscardi F (1999) Assessment of the application of baculoviruses for control of Lepidoptera.
Annu Rev Entomol 44:257–289
Nakkeeran S, Kavitha K, Mathiyazhagan S, Fernando WGD, Chandrasekar G, Renukadevi P
(2004) Induced systemic resistance and plant growth promotion by Pseudomonas chlororaphis
strain PA-23 and Bacillus subtilis strain CBE4 against rhizome rot of turmeric (Curcuma longa
L.) Can J Plant Pathol 26:417–418
Nandakumar R, Babu S, Viswanathan R, Sheela J, Raguchander T, Samiyappan R (2001) A new
bio-formulation containing plant growth promoting rhizobacterial mixture for the management
of sheath blight and enhanced grain yield in rice. BioControl 46:493–510
Naseri B, Fathipour Y, Moharramipour S, Hosseininaveh V (2009) Comparative life history and
fecundity of Helicoverpa armigera (Lepidoptera: Noctuidae) on different soybean varieties.
Entomol Sci 12(2):147–154
Neuenschwander P (2004) Harnessing nature in Africa. Nature 432:801–802
Nguyen Q, Palfreyman RW, Chan LCL, Reid S, Nielsen LK (2012) Transcriptome sequencing of
and microarray development for a Helicoverpa zea cell line to investigate in vitro insect cell-
Baculovirus interactions. PLoS One 7(5)
Nimbalkar RK, Shinde S, Tawar DS, Muley SP (2009) Response of cotton bollworm Helicoverpa
armigera (Hubner) (Lepidoptera: Noctuidae) to different insecticides in Maharashtra, India,
world. J Agric Sci 5(2):250–255
Parewa HP, Yadav J, Rakshit A, Meena VS, Karthikeyan N (2014) Plant growth promoting rhizo-
bacteria enhance growth and nutrient uptake of crops. Agric Sustain Dev 2(2):101–116
Patnaik HP, Lenka D (2000) Damage potential of Helicoverpa armigera in linseed (Linum usitatis-
simum). Indian J Agric Sci 70:197–198
Pawar CS, Borikar PS (2005) Microbial options for management of Helicoverpa armigera
(Hubner). pp 193–231
Piel J (2004) Metabolites from symbiotic bacteria. Nat Prod Rep 21:519–538
Prakash S, Verma JP (2016) Global perspective of potash for fertilizer production. In: Meena VS,
Maurya BR, Verma JP, Meena RS (eds) Potassium solubilizing microorganisms for sustainable
agriculture. Springer, New Delhi, pp 327–331. doi:10.1007/978-81-322-2776-2_23
Prasad A, Wadhwani Y, Jain M, Vyas L (2007) Pathological alteration in the protein content of
Helicoverpa armigera (Hubner) induced by Bacillus thuringiensis, npv and neem treatments.
J Herb Med Toxicol 1(1):51–53
Priyadharsini P, Muthukumar T (2016) Interactions between arbuscular mycorrhizal fungi and
potassium-solubilizing microorganisms on agricultural productivity. In: Meena VS, Maurya
BR, Verma JP, Meena RS (eds) Potassium solubilizing microorganisms for sustainable agricul-
ture. Springer, New Delhi, pp 111–125. doi:10.1007/978-81-322-2776-2_8
11 Entomopathogenic Bacteria and Biorationals in Chickpea Organic Crop Protection 255
Shah PA, Goettel MS (1999) Directory of microbial control products. Society for Invertebrate
Pathology, Division of Microbial Control. http://www.sipweb.org/directory.htm.
Shanthakumar SP, Malarvannan S, Prabavathy VR, Sudha Nair (2009) Efficacy of green muscar-
dine fungus, Metarhizium anisopliae (Metsch.) against red flour beetle, Tribolium castaneum
(Herbst) (Tenebrionidae: Coleoptera). In: Booklet on the national symposium on non-chemical
insect pest management. Elite publishing house. pp 105–11 2
Shanthakumar SP, Manivannan S, Prabavathy VR (2010a) Intraspecific plasticity in egg parasitoid,
Trichogramma chilonis Ishii (Hymenoptera: Trichogrammatidae). Pest Manag Hortic Ecosyst
16(1):5–16
Shanthakumar SP, Murali PD, Malarvannan S, Prabavathy VR, Naira S (2010b) Laboratory
investigation of the potential of entomopathogenic fungi, Nomuraea rileyi against tobacco
caterpillar, Spodoptera litura Fabricius and its safety to Trichogramma sp. J Biopest 3:132–137
Sharma RP, Yadav RP (2001) Susceptibility status of Helicoverpa armigera Hüb. to some synthetic
insecticides and neem seed kernel extract as influenced by host plant. Pestic Res J 13:152–159
Sharma A, Shankhdhar D, Shankhdhar SC (2016) Potassium-solubilizing microorganisms:
mechanism and their role in potassium solubilization and uptake. In: Meena VS, Maurya BR,
Verma JP, Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture.
Springer, New Delhi, pp 203–219. doi:10.1007/978-81-322-2776-2_15
Shrivastava M, Srivastava PC, D’Souza SF (2016) KSM soil diversity and mineral solubilization,
in relation to crop production and molecular mechanism. In: Meena VS, Maurya BR, Verma JP,
Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture. Springer,
New Delhi, pp 221–234. doi:10.1007/978-81-322-2776-2_16
Sindhu SS, Parmar P, Phour M, Sehrawat A (2016) Potassium-solubilizing microorganisms
(KSMs) and its effect on plant growth improvement. In: Meena VS, Maurya BR, Verma JP,
Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture. Springer,
New Delhi, pp 171–185. doi:10.1007/978-81-322-2776-2_13
Singh SP (1994) Fifteen years of AICRP on biological control. Project directorate of biological
control. Bangalore, New Delhi. 320 pp
Singh R (1999) A serious outbreak of Helicoverpa armigera (Hübner) on strawberries in the foot-
hills of Shivaliks. Insect Environ 5:41
Singh OP (2005) Consumption pattern of insecticide in Helicoverpa armigera management in
India. In: Recent advances in Helicoverpa armigera Management. Indian Society of Pulses
Research and Development, Kanpur, pp 17–24
Singh J, Banerjee N (2008) Transcriptional analysis and functional characterization of a gene
pair encoding iron-regulated xenocin and immunity proteins of Xenorhabdus nematophila.
J Bacteriol 190:3877–3885
Singh SP, Ballal CR, Poorani J (2002) Old world boll worm Helicoverpa armigera, associated
Heliothinane and their natural enemies. Project Directorate of Biological Control, Bangalore,
India, Technical Bulletin No. 31+135pp
Singh NP, Singh RK, Meena VS, Meena RK (2015) Can we use maize (Zea mays) rhizobacteria as
plant growth promoter? Vegetos 28(1):86–99. doi:10.5958/2229-4473.2015.00012.9
Singh M, Dotaniya ML, Mishra A, Dotaniya CK, Regar KL, Lata M (2016) Role of biofertiliz-
ers in conservation agriculture. Bisht JK, Meena VS, Mishra PK, Pattanayak A Conservation
agriculture: an approach to combat climate change in Indian Himalaya. Springer Singapore,
113–134; 10.1007/978-981-10-2558-7_4.
Stackebrandt E, Goebel BM (1994) Taxonomic note: a place for DNA-DNA reassociation and 16S
rRNA sequence analysis in the present species definition in bacteriology. Int J Syst Bacteriol
44:846–849
Stock CA, Mcloughlin TJ, Klein JA, Adang M (1990) Expression of a Bacillus thuringiensis crys-
tal protein gene in Pseudomonas cepacia 526. Can J Microbiol 36:879–884
Strasser H, Vey A, Butt TM (2000) Are there any risks in using entomopathogenic fungi for pest
control, with particular reference to the bioactive metabolites of Metarhizium, Tolypocladium
and Beauveria species. Biocontrol Sci Tech 10:717–735
11 Entomopathogenic Bacteria and Biorationals in Chickpea Organic Crop Protection 257
Subramaniam Gopalakrishnan GV, Rao R, Pagidi Humayun V, Rao R, Alekhya G, Jacob S, Deepthi
K, Meesala Sree Vidya V, Srinivas LM, Rupela O (2011) Efficacy of botanical extracts and
entomopathogens on control of Helicoverpa armigera and Spodoptera litura. Afr J Biotechnol
10(73):16667–16673
Subramanian S, Mohankumar S (2006) Genetic variability of the bollworm, Helicoverpa armig-
era, occurring on different host plants. spp. J Insect Sci 6:26
Swaminathan MS (2010) From green to evergreen revolution. Indian Agriculture: performance and
emerging challenges. Chapter 1, pp 28., Academic Foundation, New Delhi, pp 7
Teotia P, Kumar V, Kumar M, Shrivastava N, Varma A (2016) Rhizosphere microbes: potassium
solubilization and crop productivity-present and future aspects. In: Meena VS, Maurya BR,
Verma JP, Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture.
Springer, New Delhi, pp 315–325. doi:10.1007/978-81-322-2776-2_22
Thlaer JO, Baghdiguian S, Boemare N (1995) Purification and characterization of Xenorhabdicin,
a phage tail-like bacteriocin, from the lysogenic strain F1 of Xenorhabdus nematophilus. Appl
Environ Microbiol 61:2049–2052
Uma Devi K, Padmavathi J, Uma Maheswara Rao C, Khan AA, Mohan M (2008) A study of host
specificity in the entomopathogenic fungus Beauveria bassiana (Hypocreales, Clavicipitaceae).
J Biocontrol Sci Tech 18(10):975–989
Vallet-Gely I, Lemaitre B, Boccard F (2008) Bacterial strategies to overcome insect defences. Nat
Rev Microbiol 6:302–313
Vasantharaj DB, Ramamurthy (2008) Biotechnological approaches in IPM and their impact on
environment. J Biopest 1:01–05
Velazquez E, Silva LR, Ramírez-Bahena MH, Peix A (2016) Diversity of potassium-solubilizing
microorganisms and their interactions with plants. In: Meena VS, Maurya BR, Verma JP,
Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture. Springer,
New Delhi, pp 99–110. doi:10.1007/978-81-322-2776-2_7
Verma R, Maurya BR, Meena VS (2014) Integrated effect of bio-organics with chemical fertilizer
on growth, yield and quality of cabbage (Brassica oleracea var capitata). Indian J Agric Sci
84(8):914–919
Verma JP, Jaiswa DK, Meena VS, Meena RS (2015a) Current need of organic farming for enhanc-
ing sustainable agriculture. J Clean Prod 102:545–547
Verma JP, Jaiswal DK, Meena VS, Kumar A, Meena RS (2015b) Issues and challenges about sus-
tainable agriculture production for management of natural resources to sustain soil fertility and
health. J Clean Prod 107:793–794
Vining LC, Kellerher WJ, Schwarting AE (1962) Oosporein production by a strain of Beauveria
bassiana originally identified as Amanita muscaria. Can J Microbiol 8:931–933
Wagenar MM, Gibson DM, Clardy J (2002) Akanthomycin, a new antibiotic pyridone from the
entomopathogenic fungus Akanthomyces gracilis. Org Lett 4:671–673
Woodring JL, Kaya HK (1988) Steinernematid and Heterorhabditid nematodes. A hand-book of
techniques, Southern Cooperative Series Bulletin, vol 331. Arkansas Agricultural Experiment
Station, Fayetteville. 30 p
Yadav BK, Sidhu AS (2016) Dynamics of potassium and their bioavailability for plant nutrition. In:
Meena VS, Maurya BR, Verma JP, Meena RS (eds) Potassium solubilizing microorganisms for
sustainable agriculture. Springer, New Delhi, pp 187–201. doi:10.1007/978-81-322-2776-2_14
Yasin M, Munir I, Faisal M (2016) Can Bacillus spp. enhance K+ uptake in crop species. In: Meena
VS, Maurya BR, Verma JP, Meena RS (eds) Potassium solubilizing microorganisms for sus-
tainable agriculture. Springer, New Delhi, pp 163–170. doi:10.1007/978-81-322-2776-2_12
Yokoyama T, Tanaka M, Hasegawa M (2004) Novel cry gene from Paenibacillus lentimorbus
strain Semadara inhibits ingestion and promotes insecticidal activity in Anomala cuprea larvae.
J Invertebr Pathol 85:25–32
Zahedi H (2016) Growth-promoting effect of potassium-solubilizing microorganisms on
some crop species. In: Meena VS, Maurya BR, Verma JP, Meena RS (eds) Potassium sol-
ubilizing microorganisms for sustainable agriculture. Springer, New Delhi, pp 31–42.
doi:10.1007/978-81-322-2776-2_3
258 S.P. Shanthakumar
Zehnder GW, Kloepper J, Yao C, Wei G (1997) Systemic resistance against beetles (Coleoptera:
Chrysomelidae) by plant growth – promoting rhizobacteria. J Econ Entomol 90:91–39
Zhang J, Hodgman TC, Krieger L, Schnetter W, Schairer HU (1997) Cloning and analysis of the
first cry gene from B. popilliae. J Bacteriol 179:4336–4341
Zhou WW, Liang QM, Xu Y, Gurr GM, Bao YY, Geoff M, Bao GY-Y, Zhou X-P, Zhang C-X,
Cheng J, Zhu Z-R (2013) Genomic insights into the glutathione S-transferase gene family of
two rice planthoppers, Nilaparvata lugens (Stal) and Sogatella furcifera (Horvath) (Hemiptera:
Delphacidae). PLoS One 8:2
Zizka J, Weiser J (1993) Effect of beauvericin, a toxic metabolite of Beauveria bassiana, on the
ultrastructure of Culex pipiens autogenicus larvae. Cytobios 75:13–19
Bio-stimulants: An Approach Towards
the Sustainable Vegetable Production 12
Kumari Shubha, Anirban Mukherjee, Meenu Kumari,
Kalpana Tiwari, and Vijay Singh Meena
Abstract
In the context of changing climatic scenario, depletion of natural resources and
increasing hunger and malnutrition, it is necessary to ensure sustainable agricul-
tural production and resource utilization. Bio-stimulants are a viable alternative
in this condition. Bio-stimulant consists of various substances and microorgan-
isms which have found to be effective in enhancing plant growth enhance
nutrition efficiency, abiotic stress tolerance and crop quality traits. Vegetables are
a rich and relatively cheaper source of vitamins. Consumption of vegetable
provides palatability and taste, increases appetite, supplies fibre for digestion and
helps in preventing malnutrition. To enhance the sustainability of vegetable
production, bio-stimulants can be effectively used. In this chapter, status of
vegetable production worldwide and in India, categories of plant bio-stimulants
and its importance in vegetable production enhancement and factors contributing
towards the expansion of the bio-stimulants market in the world have been
discussed. The impact of PGPR carrying plentiful multifunctional plant growth-
promoting activities on vegetable crops is discussed. Furthermore, this chapter
K. Shubha (*)
ICAR-National Bureau of Plant Genetic Resources, New Delhi, Delhi 110012, India
e-mail: shubha.veg@gmail.com
A. Mukherjee
ICAR-Vivekanad Parvatiya Krishi Anusandhan Sansthan, Almora 263601, Uttarakhand, India
ICAR-Indian agricultural Research Institute, New Delhi 110012, India
M. Kumari
ICAR-IIHR, Central Horticultural Experiment Station, Bhubaneswar, Orissa 751001, India
K. Tiwari
ICAR-Indian Institute of Pulse Research, Kanpur, Uttar Pradesh 208024, India
V.S. Meena
ICAR-Vivekanad Parvatiya Krishi Anusandhan Sansthan, Almora 263601, Uttarakhand, India
Keywords
Microorganism • Horticultural crops • Biofertilizers • Nutrients • Food security
12.1 Introduction
Vegetables are a rich and relatively cheaper source of vitamins. The consumption of
vegetable provides palatability and taste, increases appetite, supplies fibre for diges-
tion and helps in preventing constipation. It plays a key role in neutralizing the acids
produced during digestion of pretentious and fatty foods and also provides valuable
roughages which help in the movement of food in the intestine.
Some of the vegetables are good sources of carbohydrates (leguminous vegetables,
potato, sweet potato, garlic, onion and fenugreek), proteins (peas, leafy vegetables,
beans and garlic), vitamin A (carrot, drumstick, tomato, leafy vegetables), vitamin
12 Bio-stimulants: An Approach Towards the Sustainable Vegetable Production 261
B (garlic, peas and tomato), vitamin C (drumstick leaves, green chillies, leafy
vegetables, cole crops and leaves of radish) and minerals (leafy vegetables,
drumstick pods). As per nutritionist, the daily requirement of vegetables is 75–125 g
of green leafy vegetables, 85 g of other vegetables and 85 g of roots and tubers with
other food.
All these vegetables are important dietary sources of micronutrients. The World
Health Organization (WHO) approximates that less fruit and vegetable intake
contributes to ~16 million disability-adjusted life years (DALYs). The DALYs is a
measure of overall disease burden expressed as the potential life lost through prema-
ture mortality and the years of productive life lost through ill-health and disability.
The approximately 1.7 million losses of life worldwide are attributable to low fruit
and vegetable consumption (FAO STAT 2014). In this regard, WHO recommended
the dietary allowance of 400 g of fruit and vegetables per day – excluding starchy
root crops for the prevention of chronic diseases like diabetes and obesity, heart
disease and cancer and for the prevention and relief from several micronutrient
deficiencies, especially in developing countries (WHO 2003). There is a huge gap
in the recommended dietary allowances and actual intakes in fruits and vegetables
worldwide due to lack of availability and accessibility for the poor masses. A low
production trend has been observed worldwide.
The annual growth rate of vegetable in the world has been declined from 6.7% in the
decade 1990–2000 to 3.4% in 2000–2011. The trend is applicable to nearly about all
countries in Asia and Pacific region except Iran (Table 12.1). Due to the increased
resource use efficiency, Iran has maintained increasing return.
India is the second largest producer of vegetables in the world (next to China)
and produces ~15% of the world’s vegetable biomass. It produces over ~90 Mt veg-
etables in ~6.2 M ha which are ~3% of the total area under cultivation in the country.
In India, more than 40 types of vegetables belonging to different groups are com-
mercially cultivated. These include solanaceous, cucurbitaceous, Allium groups,
Table 12.1 Vegetable producers and their productivity in major countries in Asia and Pacific
region
Area Production
(‘000 ha) Growth percent (‘000 tonnes) Growth percent Growth percent
Countries 2011 (2000–2011) 2011 (1990–2000) (2000–2011)
China 24,213 2.9 562,596 10.7 4.2
India 7571 3.0 105,795 4.0 3.5
Asia 40,228 2.6 802,500 8.4 4.0
Iran 885 4.3 23,764 1.2 6.7
Africaa 5388 2.1 36,846 4.7 3.4
World 56,734 2.1 1,090,425 6.7 3.4
a
Regional Office for Africa; (Source: FAO STAT 2014)
262 K. Shubha et al.
leguminous, cruciferous, root crops and leafy vegetables. Among these the five
crops, viz. potato (Solanum tuberosum), tomato (Solanum lycopersicum), onion
(Allium cepa), cabbage (Brassica oleracea var. capitata) and cauliflower (Brassica
oleracea var. botrytis), account for ~60% of the total vegetable production in the
country. Vegetables including root and tuber crops occupy an important place in the
diversification of agriculture and have played a pivotal role in food and nutritional
security of the ever-growing population of our country (Maity et al. 2013). Still, the
sector is surrounded by some issues that hinder the sustainable development of the
vegetable sector of India. The major issues are described below.
The vegetable sector suffers from a lack of availability of good quality seeds, planting
material (Shubha and Shridhar 2016; Shubha et al. 2016), fertilizer and plant protec-
tants. Availability of critical inputs is very much essential for achieving a good har-
vest. Bio-stimulants can complement the fertilizers and plant protection material.
Vegetables are highly perishable in nature, ~25–40% of the produce get wasted due
to lack of sufficient post-harvest handling measures, which cuts down the export
quantities and increases competitiveness.
12 Bio-stimulants: An Approach Towards the Sustainable Vegetable Production 263
12.4 B
io-stimulant and Its Importance in Vegetable
Production Enhancement
Bio-stimulants are mixtures of one or more things such as microorganisms, trace ele-
ments, enzymes, plant hormones and seaweed extracts rather than chemical fertiliz-
ers, which meant to correct severe nutrient deficiency. It has shown to influence several
metabolic processes such as photosynthesis, respiration, ion uptake and nucleic acid
synthesis. Bio-stimulants enhance nutrient availability, increase antioxidants, enhance
metabolism water-holding capacity and increase chlorophyll production. Besides
many advantages, the use of bio-stimulants in agricultural practices is proposed as a
safe tool to enhance the nutritional properties of food crops (Ahmad et al. 2016;
Meena et al. 2016a; Parewa et al. 2014; Prakash and Verma 2016; Kumar et al. 2016a).
Agricultural bio-stimulants comprise diverse formulations of substances, com-
pounds and microorganisms that are applied to plants or soils to improve crop
vigour, quality, yield and tolerance to abiotic stresses. It promotes plant growth and
development throughout the crop life cycle from germination to maturity in a num-
ber of established ways. Better germination and root development, greater vigour
and stress resistance, more efficient energy and nutrient uptake and transport are
possible through the use of bio-stimulants (Fig. 12.1). By improving the efficiency
of the plant’s metabolism to induce yield increases and enhance crop quality, it
helps in increasing plant tolerance to abiotic stresses and recovering from stresses;
facilitates nutrient assimilation, translocation and use; enhances quality attributes of
produce, viz. sugar content, colour and fruit seeding; and enhances soil fertility,
mainly by fostering the growth of complementary soil microorganisms (Meena
et al. 2015a, f, 2016b; Priyadharsini and Muthukumar 2016; Kumar et al. 2017;
Raghavendra et al. 2016; Dominguez-Nunez et al. 2016; Dotaniya et al. 2016;
Jaiswal et al. 2016; Jha and Subramanian 2016).
The bio-stimulants are established as ecofriendly compounds having beneficial
effects on plants (Schiavon et al. 2008). In particular, they decrease the use of chem-
ical fertilizers by escalating the amount of macro- and micronutrients taken up by
plants, positively influencing root morphology and plant growth (Nardi et al. 2009;
Ertani et al. 2013). They exhibit hormone-like activity and influence plant metabo-
lism through interacting with the biochemical processes. The manipulation of
physiological mechanisms, such as nitrogen assimilation and glycolysis are induced
to enhance plant quality parameters (Ertani et al. 2009). The mechanisms behind the
biochemical and physiological effects of bio-stimulants on vegetable are often
unidentified. It is because of the heterogeneity of the raw materials constituents
used for vegetable production (Zahedi 2016; Meena et al. 2015b, 2016c; Rawat
et al. 2016; Yasin et al. 2016; Saha et al. 2016a; Yadav and Sidhu 2016; Bahadur
et al. 2016b; Das and Pradhan 2016).
264 K. Shubha et al.
These effects are influenced by many components that may act synergistically in
different ways. Current studies suggest that the active molecules in bio-stimulants
can promote assimilation of nitrogen through stimulation of the activity and
transcription of nitrogen assimilation and Krebs’ cycle enzymes (Schiavon et al.
2008). The induction of the metabolic pathway linked with the synthesis of phenyl-
propanoids in plants treated with bio-stimulants may explain the reason behind the
plants to overcome stress situations (Ertani et al. 2013).
Pepper is an important agricultural crop known for the nutritional value. It is an
excellent source of a wide range of phytochemicals with renowned antioxidant
properties. The major antioxidant compounds include capsaicinoids, carotenoids
and phenolic compounds, particularly quercetin, flavonoids and luteolin (Howard
et al. 2000). In recent studies, the application of certain bio-stimulant to pepper
plants was found to exert affirmative effects on plant growth and yield devoid of
fruit quality degradation (Azcona et al. 2011; Meena et al. 2014a, 2016d; Saha et al.
2016b; Verma et al. 2014, 2015bVerma et al. 2014; Meena et al. 2014a).
Bio-stimulants improve nutrient availability at root rhizosphere, enhance water-
holding capacity, increase antioxidant activities and proliferate metabolism and
chlorophyll production (EBIC 2013). It distinguish themselves from traditional
crop inputs in three ways: (1) operate through different mechanisms than fertilizers,
no matter the presence of nutrients in the products, (2) act only on the plant’s vigour
and do not have any direct actions against pests or disease and (3) play a comple-
mentary role in cropping nutrition and crop protection (EBIC 2013).
Bio-stimulants have a significant impact on crop yield, thereby helping nations
cater to the surging demand for food. Keeping this in mind, the global population
explosion is one of the foremost factors driving the bio-stimulants market. The
12 Bio-stimulants: An Approach Towards the Sustainable Vegetable Production 265
Fig. 12.2 Global
bio-stimulant market share 4%
20%
42%
21%
13%
Fig. 12.3 Bio-stimulants
active ingredient sources
37%
51%
12%
Sea weeds Microbes, Chitin, plant extracts Humic and Fulvic acid
bio-stimulants can increase crop yield by 5–10% while increasing the fertilizer use
efficiency by at least 5–25% and reducing the cost of pesticides ~10–15% (EBIC
2013). The global market of bio-stimulant was mainly concentrated in
Europe (Fig. 12.2), although is projected to be spread across the world and will
reach from $1.69 billion in 2016 to $2.24 billion by 2018 (Calvo et al. 2014) and
$3.08 billion by 2022 at 10.50% CAGR (Anonymous 2016).
The bio-stimulant includes a diverse group of products, technologies that have dif-
ferent modes of action and are derived from naturally occurring microorganisms,
plant extracts or other organic matter. Worldwide the bio-stimulants’ active ingredi-
ent sources are majorly classified in three as humic acid and fulvic acid (~51%),
seaweed extracts (~37%) and microbes, and chitin and plant extract (~12%)
(Figs. 12.3). The details are presented below.
266 K. Shubha et al.
Humic substances are natural elements of the soil organic matter, consequential to
the decompositions of the animal, plant and microbial residues as a result of the
metabolic activity of soil microbes by means of these substrates. The best sources
of humic acids are found in layers of leonardite. It is an organic matter alike the soft
brown coal but differs by its degree of oxidation. Both the humic acid and fulvic
acid have been shown to possess an affirmative impact on plant growth through
providing an effective growing environment for plants by increasing surface water
infiltration, penetration and soil water-holding capacity. They also enhance the
availability of phosphorus potassium and some essential micronutrients (Gallant
2004). Humic acids play an important role in physiological and morphological
effects on plants (Eyheraguibel et al. 2008) The humic acids retain nutrient ions and
thus prevent them from leaching. They also act as a buffer for plants from too high
concentrations of fertilizer salts. Other than these the humic acids have been shown
to boost germination rates and promote greater fibrous root growth (EBIC 2013).
The bio-stimulants act in synergy with plant nutrients. The application of humic
acid and nitrogen in combination has promoted better root growth than with only
nitrogen. They also enhance chlorophyll content of plant leaves and improve stand
uniformity by influencing metabolism (Sharma et al. 2016; Verma et al. 2015a;
Meena et al. 2013b, 2015e; Teotia et al. 2016).
The fulvic acids are a kind of humic substances recognized to be powerful organic
electrolytes which help to dissolve soil minerals and metals (Huang and Deller
1970). Fulvic acids transform minerals to readily available form for easy absorption
by plants (Jackson 1993). They act more in the plant than in the soil and enhance
vitamins, coenzyme, auxin, nutrient and metabolism, which help significantly to
plant health. Furthermore, the fulvic acids help plants to resist wilting indirectly by
raising the amount of carbohydrates which results in soluble sugars accumulation in
the cell. To deal with drought stress, they increase the osmotic pressure on the cell
walls. Finally, fulvic acids help to enhance the nutrient uptake by increasing the
permeability of the cell membrane.
12.5.3 Cytokinins
Cytokinins promote cell division in plants. They have been reported to promote cell
expansion, enlarge leaf surface area that results in more chlorophyll production and
amplify photosynthesis. A low concentration of cytokinins has been used as a seed
treatment to promote lateral root development in young seedlings (Laplaze et al. 2007;
Werner et al. 2010). They also have the ability to promote nutrient translocation within
plants which is responsible for increased plant metabolism (Chang et al. 2013).
12 Bio-stimulants: An Approach Towards the Sustainable Vegetable Production 267
Protein hydrolases are actually amino acids and peptide mixtures which are obtained
by enzymatic and chemical protein hydrolysis produced from agro-industry as by-
products, from both animal (e.g. epithelial tissues, collagen) and plant sources (crop
residues) (Calvo et al. 2014; Halpern et al. 2015). In biostimulants some other
nitrogenous molecules like polyamines, betaines and ‘nonprotein amino acids’ are
there (Vranova et al. 2011). Protein hydrolysates are established for having a role in
increasing microbial biomass and activity, soil respiration and thus overall soil fer-
tility. Significant improvements in yield and quality traits have been reported in
agricultural and horticultural crops (Calvo et al. 2014). A number of commercial
products prepared from protein hydrolysates of animal and plant origins are avail-
able in the European market (Table 12.2).
Table 12.2 Examples of commercialized bio-stimulants and their use in vegetable disease
management
Products Product origin Manufacturer Crop Disease targets
Vacciplant® Laminarian extract Laboratoire Tomato Bacterial spot, bacterial
from brown algae, Goemar, speck, grey mould,
Laminaria digitata France powdery mildew,
Phytophthora blight,
early blight, Anthracnose
Egg plant Powdery mildew,
Phytophthora blight
Cucurbits Powdery mildew,
Zucchini Phytophthora blight
Cucumber
Watermelon
Melon
Leafy vegetable Downy mildew
Lettuce Grey mould
Spinach
Cabbage Downy mildew
Grey mould
Elexa® 4PDB Chitosan-based Plant Defence Cucumber, Downy mildew
natural product Boosters Inc., melon, Powdery mildew
USA pumpkin,
squash
Peas Powdery mildew
Milsana® Alcoholic extract KHH Greenhouse as Powdery mildew
from dried plant part Bioscience, well as open
of a weed giant USA; BIOFA field cucumber
knotweed (Reynoutria AG, Germany
sachalinensis)
Tomato and Powdery mildew
pepper
ChitoPlant ® Chitosan-based ChiPro GmbH, Tomato Powdery mildew Scab
natural product Germany Potato
Cucurbits Downy mildew
268 K. Shubha et al.
The farmers of developing countries will not adopt bio-stimulant and biocontrol
management practices until and unless the successes are guaranteed. The small-
scale farmers in developing countries do not always welcome the suggestion of
using alternative methods with much enthusiasm (Bashan et al. 2014). Among the
bio-based products, majority of the farmers are using bio-stimulants and biopesti-
cides. In North America, it represents ~40% of the biocontrol market as compared
with ~25% in Europe, ~20% in Asia, ~ 10% in South America and ~5% in the rest
of the world (Cox and Wong 2013).
The main reason for farmer’s laggardness about these alternative methods of
agro-ecological management is the variable efficacy in the field as compared with
conventional chemical inputs (Arora et al. 2011; Walters et al. 2013). These prod-
ucts have shown variable field performances as compared with the laboratory or in
greenhouse condition. Such inconsistency in practical conditions made this technol-
ogy not fully reliable does the farming community (Gozzo and Faoro 2013).
Naturally, the bio-stimulants depend on the concentration and interaction with
the environmental condition. As, for example, the PGPR, bacteria concentrations in
commercialized products may fall below the desired threshold level (108–1011 cells/
mL). The concentration varies under a long term or in any inadequate storage condi-
tions. Similarly, the performance of bio-stimulants greatly depends on the field
environmental condition (temperature, relative humidity, the intensity of sunlight,
concentration of UV radiation, disease pressure), crop systems (nutritional require-
ments, plant genotype, physiology) and the formulation (Walter et al. 2014).
The farmers’ adoption decision depends mainly on the label of change that they
want from the current agricultural practices and the potential of a new technology.
The level of change can be manipulated by creating a need for development through
effective utilization of extension strategies. The benefits of the technology have to
be clearly demonstrated to educational programmes, awareness campaigns obvi-
ously backed by pragmatic field results. Development of detailed package of prac-
tices about the agronomic parameters crop management techniques, the time and
frequency of dosage application procedures and other important practices (Singh
et al. 2015; Meena et al. 2013c; Bahadur et al. 2016a).
In developing countries like India, certain other things have to be considered for
successful transfer of technology. The first is the availability of the product in the
right time at the right place. It has been reported that the basic input like the seed,
fertilizer is still partially available to the farmers. In such a condition the products
like bio-stimulants have to be readily available in local markets during the pick
270 K. Shubha et al.
There are two indispensable factors for effective adoption of any particular tech-
nology knowledge about technology and compatibility of the technology with local
condition. This is true for bio-stimulants too. The knowledge factor can be taken
care of through research in applications of bio-stimulants in vegetable crops, types
of vegetable crops may be covered with the level of concentration, etc. Agronomic
field research in vegetable crop particular to Indian condition is very less which
needs to be promoted.
The compatibility of a technology depends on several other factors such as the
cost of the product, ease of application and efficiency. For that purpose, detailed
studies are required for (1) long-term survival and good shelf-life of bio-stimulant,
specificity in applications in a wide range of agroecological conditions in the veg-
etable crop, (2) cost reduction mechanisms, (3) suitable conditions dosage, concen-
tration and (4) farm machinery applications of bio-stimulants.
Presently, India does not have much production units of bio-stimulants. It majorly
depends on imports. In 2016, India has imported 492 tonnes of bio-stimulants worth
$ 15.78 Million. The United States is the largest supplier of bio-stimulants account-
ing for imports worth $ 9.75 Million followed by Mexico and Israel which exported
bio-stimulants worth $ 2.93 Million and $ 1.37 Million, respectively (Zauba.com
2016). Although very less amount is used for plant growth promotion. Efforts are
required to production, promotion and adoption of bio-stimulants. In such a condi-
tion, public-private partnership and convergence approach are required (Mukherjee
et al. 2012a; Mukherjee and Maity 2015).
12 Bio-stimulants: An Approach Towards the Sustainable Vegetable Production 271
12.7 Conclusions
In order to enhance the sustainable food production, food and nutritional security
with a significant reduction of synthetic fertilizers, agrochemical use and environ-
mental pollution and natural resource productivity enhancement through the use of
bio-stimulants in the vegetable are essential. It promotes plant growth and develop-
ment throughout the crop life cycle from germination to maturity in a number of
established ways. Better germination and root development, greater vigour and
stress resistance, more efficient energy and nutrient uptake and transport are possi-
ble through the use of bio-stimulants. India is a larger import of plant bio-stimulants.
For better adoption of bio-stimulants, research and extension are both required.
Especially the knowledge and awareness about the bio-stimulant technology and
compatibility of the commercial product which Indian farmers’ condition are
necessary.
Acknowledgements We would like to thank the Indian Council of Agricultural Research for
providing support and facilities and reviewers and editors of the book for their valuable
comments.
References
Ahmad M, Nadeem SM, Naveed M, Zahir ZA (2016) Potassium-solubilizing bacteria and their
application in agriculture. In: Meena VS, Maurya BR, Verma JP, Meena RS (eds) Potassium
solubilizing microorganisms for sustainable agriculture. Springer, New Delhi, pp 293–313.
doi:10.1007/978-81-322-2776-2_21
Anonymous (2016) Bio-stimulants Market Worth USD 3.08 Billion by 2022 – Scalar Market
Research. http://www.cleveland19.com/story/33459526/Bio-stimulants-market-worth-usd-
308-billion-by-2022-scalar-market-research. Viewed on 18–11-2016
Arora NK, Khare E, Maheshwari DK (2011) Plant growth promoting rhizobacteria: constraints
in bioformulation, commercialization, and future strategies. In: Maheshwari DK (ed) Plant
growth and health promoting bacteria. Springer, Berlin, pp 97–116
272 K. Shubha et al.
Kumar A, Patel JS, Bahadur I, Meena VS (2016b) The molecular mechanisms of KSMs for
enhancement of crop production under organic farming. In: Meena VS, Maurya BR, Verma JP,
Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture. Springer,
New Delhi, pp 61–75. doi:10.1007/978-81-322-2776-2_5
Kumar A, Maurya BR, Raghuwanshi R, Meena VS, Islam MT (2017) Co-inoculation with
Enterobacter and Rhizobacteria on yield and nutrient uptake by wheat (Triticum aestivum L.)
in the alluvial soil under indo-gangetic plain of India. J Plant Growth Regul. doi:10.1007/
s00344-016-9663-5
Laplaze L, Benkova E, Casimiro I et al (2007) Cytokinins act directly on lateral root founder cells
to inhibit root initiation. Plant Cell 19:3889–3900
Lim JH, Kim SD (2013) Induction of drought stress resistance by multi-functional PGPR Bacillus
licheniformis K11 in pepper. Plant Pathol J 29:201–208
Maity A, Mukherjee A, Pramanik P, Shubha (2013) Shifting towards vegetable from boro rice
cultivation in Purba Medinipur district of West Bengal. Bioinfolet 10(3A):876–878
Mangmang JS, Deaker R, Rogers G (2015) Germination characteristics of cucumber influenced
by plant growth promoting Rhizobacteria. Int J Veg Sci http://dx.doi.org/10.1080/19315260.
2014.938850
Masood S, Bano A (2016) Mechanism of potassium solubilization in the agricultural soils by the
help of soil microorganisms. In: Meena VS, Maurya BR, Verma JP, Meena RS (eds) Potassium
solubilizing microorganisms for sustainable agriculture. Springer, New Delhi, pp 137–147.
doi:10.1007/978-81-322-2776-2_10
Maurya BR, Meena VS, Meena OP (2014) Influence of Inceptisol and Alfisol’s potassium solubi-
lizing bacteria (KSB) isolates on release of K from waste mica. Vegetos 27:181–187
Meena OP, Maurya BR, Meena VS (2013a) Influence of K-solubilizing bacteria on release of
potassium from waste mica. Agric Sust Dev 1:53–56
Meena VS, Maurya BR, Bohra JS, Verma R, Meena MD (2013b) Effect of concentrate manure
and nutrient levels on enzymatic activities and microbial population under submerged rice in
alluvium soil of Varanasi. Crop Res 45(1, 2 & 3):6–12
Meena VS, Maurya BR, Verma R, Meena RS, Jatav GK, Meena SK, Meena SK (2013c) Soil
microbial population and selected enzyme activities as influenced by concentrate manure and
inorganic fertilizer in alluvium soil of Varanasi. Bioscan 8(3):931–935
Meena VS, Maurya BR, Bahadur I (2014a) Potassium solubilization by bacterial strain in waste
mica. Bang J Bot 43:235–237
Meena VS, Maurya BR, Verma JP (2014b) Does a rhizospheric microorganism enhance K+ avail-
ability in agricultural soils? Microbiol Res 169:337–347
Meena RS, Meena VS, Meena SK, Verma JP (2015a) The needs of healthy soils for a healthy
world. J Clean Prod 102:560–561
Meena RS, Meena VS, Meena SK, Verma JP (2015b) Towards the plant stress mitigate the agricul-
tural productivity: a book review. J Clean Prod 102:552–553
Meena VS, Maurya BR, Meena RS (2015c) Residual impact of wellgrow formulation and NPK on
growth and yield of wheat (Triticum aestivum L.) Bangladesh J Bot 44(1):143–146
Meena VS, Maurya BR, Verma JP, Aeron A, Kumar A, Kim K, Bajpai VK (2015d) Potassium
solubilizing rhizobacteria (KSR): isolation, identification, and K-release dynamics from waste
mica. Ecol Eng 81:340–347
Meena VS, Meena SK, Verma JP, Meena RS, Ghosh BN (2015e) The needs of nutrient use efficiency
for sustainable agriculture. J Clean Prod 102:562–563. doi:10.1016/j.jclepro.2015.04.044
Meena VS, Verma JP, Meena SK (2015f) Towards the current scenario of nutrient use efficiency in
crop species. J Clean Prod 102:556–557. doi:10.1016/j.jclepro.2015.04.030
Meena RK, Singh RK, Singh NP, Meena SK, Meena VS (2016a) Isolation of low temperature sur-
viving plant growth-promoting rhizobacteria (PGPR) from pea (Pisum sativum L.) and docu-
mentation of their plant growth promoting traits. Biocatal Agric Biotechnol 4:806–811
Meena RS, Bohra JS, Singh SP, Meena VS, Verma JP, Verma SK, Sihag SK (2016b) Towards the
prime response of manure to enhance nutrient use efficiency and soil sustainability a current
need: a book review. J Clean Prod 112(1):1258–1260
12 Bio-stimulants: An Approach Towards the Sustainable Vegetable Production 275
Meena SK, Rakshit A, Meena VS (2016c) Effect of seed bio-priming and N doses under varied
soil type on nitrogen use efficiency (NUE) of wheat (Triticum aestivum L.) under greenhouse
conditions. Biocatal Agric Biotechnol 6:68–75
Meena VS, Bahadur I, Maurya BR, Kumar A, Meena RK, Meena SK, Verma JP (2016d) Potassium-
solubilizing microorganism in evergreen agriculture: an overview. In: Meena VS, Maurya BR,
Verma JP, Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture.
Springer, New Delhi, pp 1–20. doi:10.1007/978-81-322-2776-2_1
Meena VS, Meena SK, Bisht JK, Pattanayak A (2016e) Conservation agricultural practices in
sustainable food production. J Clean Prod 137:690–691
Mukherjee A, Maity A (2015) Public–private partnership for convergence of extension services in
Indian agriculture. Curr Sci 109(9):1557–1563
Mukherjee A, Bahal R, Burman RR, Dubey SK, Jha GK (2011) Effectiveness of Tata Kisan
Sansar in technology advisory and delivery services in Uttar Pradesh. Indian Res J Ext Educ
11(3):8–13
Mukherjee A, Bahal R, Burman RR, Dubey SK (2012a) Factors contributing farmers’ association
in Tata Kisan Sansar: a critical analysis. Indian Res J Ext Educ 12(2):81–86
Mukherjee A, Bahal R, Burman RR, Dubey SK, Jha GK (2012b) Constraints in privatized agricul-
tural technology delivery system of Tata Kisan Sansar. J Glob Commun 5(2):155–159
Mukherjee A (2015) Prioritization of problems in integrated agriculture: a case of Rampur village
in sub humid region of Eastern India. Indian Res J Ext Edu 15(1):53–59
Nardi S, Carletti P, Pizzeghello D, Muscolo A (2009) Biological activities of humic substances. In:
Senesi N, Xing B, Huang PM (eds) Biophysico-chemical processes involving natural nonliving
organic matter in environmental systems. Wiley, Hoboken, pp 305–339
Parewa HP, Yadav J, Rakshit A, Meena VS, Karthikeyan N (2014) Plant growth promoting rhizo-
bacteria enhance growth and nutrient uptake of crops. Agric Sustain Dev 2(2):101–116
Prakash S, Verma JP (2016) Global perspective of potash for fertilizer production. In: Meena VS,
Maurya BR, Verma JP, Meena RS (eds) Potassium solubilizing microorganisms for sustainable
agriculture. Springer, New Delhi, pp 327–331. doi:10.1007/978-81-322-2776-2_23
Priyadharsini P, Muthukumar T (2016) Interactions between arbuscular mycorrhizal fungi and
potassium-solubilizing microorganisms on agricultural productivity. In: Meena VS, Maurya
BR, Verma JP, Meena RS (eds) Potassium solubilizing microorganisms for sustainable agricul-
ture. Springer, New Delhi, pp 111–125. doi:10.1007/978-81-322-2776-2_8
Raghavendra MP, Nayaka NC, Nuthan BR (2016) Role of rhizosphere microflora in potas-
sium solubilization. In: Meena VS, Maurya BR, Verma JP, Meena RS (eds) Potassium
solubilizing microorganisms for sustainable agriculture. Springer, New Delhi, pp 43–59.
doi:10.1007/978-81-322-2776-2_4
Rawat J, Sanwal P, Saxena J (2016) Potassium and its role in sustainable agriculture. In: Meena
VS, Maurya BR, Verma JP, Meena RS (eds) Potassium solubilizing microorganisms for sus-
tainable agriculture. Springer, New Delhi, pp 235–253. doi:10.1007/978-81-322-2776-2_17
Saha M, Maurya BR, Meena VS, Bahadur I, Kumar A (2016a) Identification and characterization
of potassium solubilizing bacteria (KSB) from Indo-Gangetic Plains of India. Biocatal Agric
Biotechnol 7:202–209
Saha M, Maurya BR, Bahadur I, Kumar A, Meena VS (2016b) Can potassium-solubilising bacteria
mitigate the potassium problems in India? In: Meena VS, Maurya BR, Verma JP, Meena RS
(eds) Potassium solubilizing microorganisms for sustainable agriculture. Springer, New Delhi,
pp 127–136. doi:10.1007/978-81-322-2776-2_9
Schiavon M, Ertani A, Nardi S (2008) Effects of an alfalfa protein hydrolysate on the gene expres-
sion and activity of enzymes of the tricarboxylic acid (TCA) cycle and nitrogen metabolism in
Zea mays L. J Agric Food Chem 56:11800–11808
Sharma A, Shankhdhar D, Shankhdhar SC (2016) Potassium-solubilizing microorganisms:
mechanism and their role in potassium solubilization and uptake. In: Meena VS, Maurya BR,
Verma JP, Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture.
Springer, New Delhi, pp 203–219. doi:10.1007/978-81-322-2776-2_15
276 K. Shubha et al.
Shrivastava M, Srivastava PC, D’Souza SF (2016) KSM soil diversity and mineral solubilization,
in relation to crop production and molecular mechanism. In: Meena VS, Maurya BR, Verma JP,
Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture. Springer,
New Delhi, pp 221–234. doi:10.1007/978-81-322-2776-2_16
Shubha KS (2016) Generation mean analysis for pod yield and its associated traits in garden pea
(Pisum sativum L.) Veg Sci 42(2):43–46
Shubha KS, Choudhary H, Dubey SC, Sharma RK (2016) Identification of resistant sources and
inheritance of fusarium wilt resistance in garden pea. Ind J Hortic 73(3):356–361
Sindhu SS, Parmar P, Phour M, Sehrawat A (2016) Potassium-solubilizing microorganisms
(KSMs) and its effect on plant growth improvement. In: Meena VS, Maurya BR, Verma JP,
Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture. Springer,
New Delhi, pp 171–185. doi:10.1007/978-81-322-2776-2_13
Singh NP, Singh RK, Meena VS, Meena RK (2015) Can we use maize (Zea mays) rhizobacteria as
plant growth promoter? Vegetos 28(1):86–99. doi:10.5958/2229-4473.2015.00012.9
Singh M, Dotaniya ML, Mishra A, Dotaniya CK, Regar KL, Lata M (2016) Role of biofertil-
izers in conservation agriculture. In: Bisht JK, Meena VS, Mishra PK, Pattanayak A (eds)
Conservation agriculture: an approach to combat climate change in Indian Himalaya. Springer,
Singapore, pp 113–134. doi:10.1007/978-981-10-2558-7_4
Tanwar A, Aggarwal A, Parkash V (2014) Effect of bioinoculants and superphosphate fertilizer on
the growth and yield of broccoli (Brassica oleracea L. var. italica Plenck). N Z J Crop Hortic
Sci 42(4):288–302
Teotia P, Kumar V, Kumar M, Shrivastava N, Varma A (2016) Rhizosphere microbes: potassium
solubilization and crop productivity-present and future aspects. In: Meena VS, Maurya BR,
Verma JP, Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture.
Springer, New Delhi, pp 315–325. doi:10.1007/978-81-322-2776-2_22
Vacheron J et al (2013) Plant growth-promoting rhizobacteria and root system functioning. Front
Plant Sci 4:1–19
Velazquez E, Silva LR, Ramírez-Bahena MH, Peix A (2016) Diversity of potassium-solubilizing
microorganisms and their interactions with plants. In: Meena VS, Maurya BR, Verma JP,
Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture. Springer,
New Delhi, pp 99–110. doi:10.1007/978-81-322-2776-2_7
Verma R, Maurya BR, Meena VS (2014) Integrated effect of bio-organics with chemical fertilizer
on growth, yield and quality of cabbage (Brassica oleracea var capitata). Indian J Agric Sci
84(8):914–919
Verma JP, Jaiswa DK, Meena VS, Meena RS (2015a) Current need of organic farming for enhanc-
ing sustainable agriculture. J Clean Prod 102:545–547
Verma JP, Jaiswal DK, Meena VS, Kumar A, Meena RS (2015b) Issues and challenges about sus-
tainable agriculture production for management of natural resources to sustain soil fertility and
health. J Clean Prod 107:793–794
Vranova V, Rejsek K, Skene KR, Formanek P (2011) Non-protein amino acids: plant, soil and
ecosystem interactions. Plant Soil 342:31–48
Walter DR, Newton AC, Lyon GD (eds) (2014) Induced resistance for plant defence: a sustainable
approach to crop protection, 2nd edn. Wiley-Blackwell, Oxford
Walters DR, Ratsep J, Havis ND (2013) Controlling crop diseases using induced resistance: chal-
lenges for the future. J Exp Bot 64(5):1263–1280
Wang CJ, Yang W, Wang C, Gu C, Niu DD, Liu HX, Wang YP, Guo JH (2012) Induction of drought
tolerance in cucumber plants by a consortium of three plant growth-promoting Rhizobacterium
strains. PLoS One 7:1–10
Werner T, Nehnevajova E, Köllmer I, Novák O, Strnad M, Krämer U, Schmülling T (2010) Root-
specific reduction of cytokinin causes enhanced root growth, drought tolerance, and leaf min-
eral enrichment in Arabidopsis and tobacco. Plant Cell 22:3905–3920
Wezel A et al (2014) Agroecological practices for sustainable agriculture. A review. Agron Sustain
Dev 34(1):1–20
12 Bio-stimulants: An Approach Towards the Sustainable Vegetable Production 277
WHO (2003) Diet, nutrition and the prevention of chronic diseases: report of a Joint WHO/FAO
Expert Consultation, WHO Technical Report Series, vol 916. World Health Organization,
Geneva
Yadav BK, Sidhu AS (2016) Dynamics of potassium and their bioavailability for plant nutrition. In:
Meena VS, Maurya BR, Verma JP, Meena RS (eds) Potassium solubilizing microorganisms for
sustainable agriculture. Springer, New Delhi, pp 187–201. doi:10.1007/978-81-322-2776-2_14
Yasin M, Munir I, Faisal M (2016) Can Bacillus spp. enhance K+ uptake in crop species. In: Meena
VS, Maurya BR, Verma JP, Meena RS (eds) Potassium solubilizing microorganisms for sus-
tainable agriculture. Springer, New Delhi, pp 163–170. doi:10.1007/978-81-322-2776-2_12
Yildirim E, Karlidag H, Turan M, Dursun A, Goktepe F (2011) Growth, nutrient uptake and
yield promotion of broccoli by plant growth promoting rhizobacteria with manure. Hort Sci
46:932–936
Zahedi H (2016) Growth-promoting effect of potassium-solubilizing microorganisms on
some crop species. In: Meena VS, Maurya BR, Verma JP, Meena RS (eds) Potassium sol-
ubilizing microorganisms for sustainable agriculture. Springer, New Delhi, pp 31–42.
doi:10.1007/978-81-322-2776-2_3
Zauba.com (2016) Analysis of imports of bio-stimulants. Available at https://www.zauba.com/
importanalysis-Bio-stimulants-report.html. Viewed on 02-12-2016
Efficacy of Biological Soil Amendments
and Biocontrol Agents for Sustainable 13
Rice and Maize Production
Manoj Kaushal and Suhas P. Wani
Abstract
Exploiting the agroecosystem services of soil microbes appears as a promising
effective approach to alleviate the negative impacts on soil systems and crop
production. Lack of soil organic matter (SOM) is a prevalent feature of degraded
soils. Different biological soil amendments, such as organic manure, compost,
vermicompost, indigenous microbes, and crop residues, are widely used in rec-
lamation of degraded soils. The biological soil amendments also furnish a valu-
able source of fertilizer for growing rice and maize and also boost physicochemical
and biological parameters of soil such as water holding capacity, moisture con-
tent, electrical conductivity, organic carbon content, and population of beneficial
microbes which directly correlates to soil health and fertility. However, effi-
ciency of amendments applied based on variety of factors including the composi-
tion and characteristics, soil microflora, and environmental conditions can
accelerate initial reclamation and lead to self-sustaining primary productivity of
crops. On the other hand, high cost of chemical fertilizers, pesticides, and other
agricultural inputs and their harmful environmental legacy have encouraged
researchers to explore the use of microbial-mediated amendments to play a cen-
tral role in raising productivity and inhibition/suppression of pathogenic popula-
tion below levels at which they cause economic and other effects to the crops as
well as the environment. Biological control can be achieved through one or more
mechanisms, viz., antibiosis, competition for nutrients and/space, induced resis-
tance, plant growth promotion, and rhizosphere colonization ability. Potent bio-
control agents such as Bacillus sp., Pseudomonas sp., and Tricoderma sp. prove
to be very promising in controlling soilborne diseases of rice and maize crops
employing both antibiosis and induction of host resistance. Determination of the
Keywords
Biological soil amendments • Biocontrol mechanisms • Intermediate metabolites
• Induced systemic resistance • Rhizobacteria • Sustainable agriculture
13.1 Introduction
Rice (Oryza sativa L.), a widely grown crop in tropical and subtropical regions, is
the most important staple foods for ~50% of the world’s human population and
requires ~10 Mt. of N fertilizers every year (Zhang et al. 2014). N source supplied
to crop is usually by commercially available N fertilizers such as urea, which results
in a substantial amount of loss through different mechanisms and thus becomes one
of the predominant factors for environmental hazards and deterioration of soil fertil-
ity as well (Meena et al. 2013a; Bahadur et al. 2014; Maurya et al. 2014; Jat et al.
2015; Kumar et al. 2015, 2016b; Ahmad et al. 2016).
The sustainable maize production systems also face major challenges of appro-
priate management of N fertilizers to maximize economic return while minimizing
environmental pollution. Maize, grown in rotation with legume crops (alfalfa or
soybeans), often yields more, so organic N in crop residues derived from biological
N2 fixation by legume crops has been an effective alternative to meet this challenge
(Raaijmakers and Lugtenberg 2013). Increase maize yield from ~5 to 20% attrib-
uted to the beneficial effect of crop rotation including improved soil physical prop-
erties (Barea 2015), a better balance of plant nutritional factors, increased root
activity, a shift in soil mycorrhizal populations, lessen disease severity (Reid et al.
2001), and magnified seasonal N mineralization (Browne et al. 2013). A large effort
has been devoted to the studies of legume effects on subsequent cereal crops grown
in rotation systems (Barea et al. 2013a; Lugtenberg 2015). The use of biological soil
amendments such as manure and compost improves soil fertility and soil quality as
well as enhances the population of beneficial microbes in rice and maize fields.
Biological soil amendments of animal origin such as manure and compost that
include animal waste and human waste serve as a potential vector for soil- and root-
borne pathogens and also critical in sustainable farming systems and certified
organic production. Addition of dairy farm manure provides partial or complete N
and other nutrient elements, as well as increases soil N release through seasonal N
mineralization and availability to the plant (Zolla et al. 2013). However, nutrient (N)
availability from manure varied greatly due to source of manure, method of storage
(Eghball 2000), and time and method of application. Manure application usually
increases crop yield due to plant nutrient availability (Ma et al. 1999b) and improved
13 Efficacy of Biological Soil Amendments and Biocontrol Agents for Sustainable… 281
Table 13.1 List of major rice diseases and their causal organisms
Disease Causal organism Biocontrol agents
Bacterial blight and leaf Xanthomonas oryzae Streptomyces sp.
streak
Blast Pyricularia grisea P. fluorescens
Brown spot Bipolaris oryzae P. aeruginosa
Crown sheath rot Gaeumannomyces graminis P. fluorescens
False smut Ustilaginoidea virens Pseudomonas sp.
Foot rot Erwinia chrysanthemi Azotobacter sp.
Grain rot Burkholderia glumae Bacillus sp.
Leaf scald Rhynchosporium oryzae Pseudomonas sp.
Leaf smut Entyloma oryzae Pseudomonas sp.
Pecky rice (kernel spotting) Curvularia sp.; Fusarium sp. Pseudomonas sp.
Root rots Pythium spinosum P. fluorescens
Seedling blight Sclerotium rolfsii B. glumae
Sheath blight Rhizoctonia solani P. fluorescens
Sheath brown rot Pseudomonas fuscovaginae B. amyloliquefaciens
Sheath rot Sarocladium oryzae P. aeruginosa
Sheath spot Rhizoctonia oryzae Pseudomonas sp.
Stem rot Sclerotium oryzae P. fluorescens
Seed-rot and seedling disease Fusarium sp.; Pythium sp. Pseudomonas spp.
soil structure (Spence and Bais 2013). The N fertilizer application can be reduced
by the use of biological N2 fixation technology which further minimizes environ-
mental risks (Raimam et al. 2007).
N-fixing bacteria may be free-living or symbiotically associated to rice plants
mostly species of Alcaligenes, Azospirillum, Bacillus, Enterobacter, Herbaspirillum,
Klebsiella, Pseudomonas, and Rhizobium (Hirsch et al. 2013b). The most serious
constraint on high productivity of rice (Table 13.1) and maize (Table 13.2) crop is
the diseases caused by fungi, bacteria, viruses, or nematodes (Manandhar et al.
1998) causing annual yield losses in very high amount. Beyond good agronomic
practices (resistant cultivars and application of pesticides), a wide variety of
approaches may be used to prevent, mitigate, or control rice diseases such as heavily
use of chemical fertilizers and pesticides.
However, many resistant cultivars have very short life span due to the breakdown
of the resistance in high pathogenic variability of the pathogens. Also, the environ-
mental pollution caused by excessive use of agrochemicals as well as strict regula-
tions on chemical pesticide has led to considerable changes in attitudes of people’s
toward the use of more pesticides in rice and maize crops. Among the alternatives,
a variety of biological controls are available for use in rice as well as maize crops,
but their effective adoptions will require a greater understanding of the complex
interactions among plants, soil, and microbes (Meena et al. 2016a; Parewa et al.
2014; Prakash and Verma 2016; Jaiswal et al. 2016; Jha and Subramanian 2016;
Kumar et al. 2016a). This article presents an advanced survey of biological soil
282 M. Kaushal and S.P. Wani
Table 13.2 List of major maize diseases and their causal organisms
Disease Causal organism Biocontrol agent
Anthracnose leaf blight Colletotrichum graminicola Torulaspora globosa
Aspergillus ear Aspergillus flavus Streptomyces sp.
Bacterial leaf spot Xanthomonas campestris B. subtilis
Bacterial stalk and top rot Erwinia carotovora B. subtilis
Bacterial stripe Pseudomonas andropogonis Microbacterium maritypicum
Black bundle disease Cephalosporium acremonium Glomus fasciculatum
Charcoal rot Macrophomina phaseolina Trichoderma sp.
Cladosporium rot Cladosporium herbarum Trichoderma sp.
Fusarium ear Fusarium subglutinans T. harzianum
Holcus spot Pseudomonas syringae M. maritypicum
Leaf spots, minor Alternaria alternata Pseudomonas sp.
Penicillium ear rot Penicillium sp. Trichoderma sp.
Seedling blight Gibberella fujikuroi Bacillus sp.
Sheath spot Rhizoctonia solani T. harzianum
Smut Ustilago maydis T. viride
Stalk rot Physoderma maydis T. machilonis
Stewart’s disease Erwinia stewartii M. maritypicum
Yellow leaf blight Phyllosticta maydis Pantoea ananas
amendments and biocontrol agents applied to the suppression of plant diseases for
sustainable rice and maize production (Zancarini et al. 2013).
Any material of plant or animal origin added to the soil to brush up its physico-
chemical and biological properties in relation to plant growth or ameliorate the
WHC, avoiding contamination of pathogens, is considered as biological soil amend-
ments (BSAs). The BSAs consist of whole or a part of materials of animal/plant
origin, such as manure, blood meal, and fish emulsion and human waste, compost,
or table waste, used alone or in consortium. Moreover, BSAs increase soil microbial
activity, uptake of nutrients, decomposition rate, and crop vigor and yield while
reducing fertilizer need leading to maximize yields (Fig. 13.1). However, there is
still lack of information available with regard to utility of BSAs, organic amend-
ments, growth regulators, and other biological stimulants. BSAs may be treated or
untreated and transform depleted soil into healthy soil (Zolla et al. 2013; Meena
et al. 2015a, f, 2016b; Priyadharsini and Muthukumar 2016; Kumar et al. 2017;
Dotaniya et al. 2016).
Treated BSAs are substitute in which microbial/bacterial inoculums are added
that have been isolated from highly fertile soils and exhibit optimum abilities to
produce organic nutrients and beneficial enzymes. This can increase availability of
labile organic carbon and more accumulation of soil organic carbon such as azolla-
algal biofertilizers which increases accumulation of ferric iron oxide in soil and
13 Efficacy of Biological Soil Amendments and Biocontrol Agents for Sustainable… 283
Crop Health
Fig. 13.1 An overview of biological soil amendments (BSAs) and their role toward the soil
sustainability
ultimately increases soil redox value and soil porosity in rice cropping system.
Azolla anabaena treatment also diminishes CH4 emission rate during the rice grow-
ing seasons due to the lower C/N ratio (14:1) and maximizes cellulose and lignin
content. Agnihotri et al. (1999) also reported that rice straw application before
flooding and the biofertilizers after flooding enhanced CH4 efflux from rice fields,
while CH4 fluxes were decreased by composts of cow dung. Composted livestock
manure application in rice field increased rice yield by ~30% and dwindle CH4
emissions 56% per unit of grain yield compared to the non-fertilized control treat-
ment. Availability of major nutrients such as N, P, K, Ca, Mg, Si, soluble Fe, etc. can
also be increased to rice and maize plants by adding BSAs, organic materials, inor-
ganic fertilizer, and other amendments (Ali et al. 2012), which further stimulated
plant growth and yield (Hu and Wang 1995; Lee et al. 2004).
284 M. Kaushal and S.P. Wani
Overall BSAs such as straw compost, cattle manure compost, green plant bio-
mass, and azolla anabaena improve soil quality parameters such as porosity, total
nitrogen, MBC, MBN, SOM, CEC, and nutrient availability to plants in paddy and
maize fields alone or as consortium or in combination with other chemical fertiliz-
ers. In addition to nutrients released from the added BSAs, they also accelerate the
rice and maize plant growth, photosynthetic carbon assimilation and root exudation,
and microbial activity and increased the MBC that eventually resulted in organic C
accumulation in soil (Ali et al. 2012). Conclusively, to minimize the use of conven-
tional NPK fertilizers, reducing CH4 emissions and sustaining rice productivity,
BSAs could be introduced in rice and maize cropping systems.
Naturally occurring soil microbes are nature’s most constructive fertilizer producers
and serve as supreme agents of BSAs. Thus, BSAs are adopted for their multiple
benefits to sustained plant growth and overall soil productivity and disease suppres-
sion. They are also effective in different soil conditions and provide unique propen-
sity to fix both atmospheric and chemical nitrogen and thus selected for utmost
adaptability. These rhizospheric bacteria are ruling in this category due to their natu-
ral enzyme production capabilities which transform insoluble form of phosphorus
and potassium to soluble form. Besides they also produce antibiotics and other plant
hormones that stimulate root and shoot growth of rice plants. These microbes are
isolated, screened, and grown in the laboratory under aseptic conditions to provide
a pure microbial culture.
Thereafter, pure microbial culture is combined with other BSAs and applied to
soil so that indigenous bacteria multiply, grow, and get adapted to conditions rap-
idly. BSAs improve the biological health of soil, thereby increasing nutrient uptake
by 150–250% and reducing volatilization (gaseous escape) and leaching run off and
seepage. Increasing BSAs of a soil makes the soil more permeable to absorb addi-
tional water and healthy populations of microbes metabolizing nutrients, making
them available to plants (Duarte et al. 2006).
13.2.2 Humus
Decomposed parts of plant and animal organic matter within the soil that resists
further decomposition are called humus, and the process of formation is called
humification. Humus profoundly improves soil structure and permeability, enhances
water holding capacity of soil, and naturally chelates unavailable micronutrients.
Active humus within the soil is created by both biological and chemical processes
and is composed largely of three organic acids: humic acid, humin, and fulvic acid
(Raghavendra et al. 2016; Zahedi 2016; Meena et al. 2015b, 2016c; Rawat et al.
2016; Yasin et al. 2016; Das and Pradhan 2016; Dominguez-Nunez et al. 2016).
13 Efficacy of Biological Soil Amendments and Biocontrol Agents for Sustainable… 285
These small organic materials join together to form complex chemical structures
called polymers. Chemically, these are polyphenolic acids, and synthesis of these
polymers results from primary transformative mechanism, i.e., release of enzymes
from different microbes. Humus is considered as “Black Gold” of soil as it darkens
soil color and increases absorption of organic compounds that aid plant nutrition,
buffering capacity of soil, thereby helping to stabilize the soil’s pH and thus plants
for absorbing mineral nutrients as well in cleaning of toxic materials in the soil
(Mercado-Blanco 2015).
13.2.3 Biostimulants
This category of BSAs is derived from plant and animal soluble comprising simple
and complex sugars, amino acids, carbohydrates, peptides, lignin, and cellulose.
They help to break soil compaction, improve CEC, and serve as an immediate
source of energy for soil microbes and plants.
Plant- Plant-pathogen
rhizobacterial interactions
interactions
ISR SAR
Biocontrol
Direct action Indirect action
Fig. 13.2 Different mechanisms carried out by BCAs leading to biological control (mainly rice
and maize crops)
13.3.1 Antagonism
Interaction involving physical contact of the pathogen with BCAs and other organ-
isms would be required to exert a suppressive effect referred to as direct antago-
nism. Indirect antagonisms do not involve targeting a pathogen by BCAs, for
example, stimulation of plant host defense pathways by nonpathogenic ones.
288 M. Kaushal and S.P. Wani
13.3.2 Mutualism
13.3.3 Commensalism
Some microbes live in a symbiotic interaction with plants, where one gets the ben-
efit and the other is neither harmed nor benefitted creating challenges to soilborne
pathogens. Increasing iron uptake efficiency of the commensal microbes is also an
important factor to colonize plant roots which leads to the displacement of patho-
gens from infected areas/sites.
13.3.4 Protocooperation
Both partners count in association do not depend on each other for survival. Some
microbes isolated and classified as BCAs are involved in protocooperation depend-
ing on environmental conditions leading to disease suppression.
13.3.5 Competition
The rhizosphere region of plant secretes root exudates such as flavonoids, sugars,
carbohydrates, proteins, amino acids, etc. for which competition exists between
pathogens and BCAs, thus becoming an important factor for limiting disease inci-
dence and severity. For example, soilborne pathogens (Fusarium and Pythium)
infect through mycelial contact and are more susceptible to competition than other
microbes. These microbes produce secondary metabolites that suppress soilborne
fungal pathogens. Another example of biological control is iron limitation in the
rhizosphere region which depends on soil pH to support the growth of microorgan-
isms, generally iron (ferric form) which is insoluble in water (pH 7.4) needed with
concentrations of 10−6 M. However, highly oxidized and aerated soil contains much
lower concentration (10−18 M) (Barea and Richardson 2015).
13.3.6 Antagonism
suppression of plant pathogens directly by lyse cell walls and act to decompose
plant residues. For example, Lysobacter and Myxobacteria are known to produce
lytic enzymes and thus suppress fungal plant pathogens. Serratia marcescens pro-
duce chitinase for the control of Sclerotium rolfsii (Ordentlich et al. 1988), and a
b-1, 3-glucanase significantly contributes to biocontrol activities of Lysobacter
enzymogenes strain C3. Microbes also produce hydrogen cyanide (HCN) which
effectively blocks the cytochrome oxidase pathway and is highly toxic to all aerobic
microorganisms and thus suppresses plant pathogens. HCN production by fluores-
cent pseudomonads is involved in the suppression of root pathogens. P. fluorescens
CHA0 produces antibiotics, siderophores, and HCN and inhibits the growth of
Thielaviopsis basicola (Olivares et al. 2013). Enterobacter cloacae suppress
Pythium ultimum-induced damping-off by producing volatile compounds such as
ammonia.
13.3.7 Predation
compounds leads to the failure to elicit ISR in certain hosts (Van Loon 2007). P.
putida WCS358r and P. fluorescensWCS374r act differently depending on the plant
species: in Arabidopsis, WCS358r elicits ISR, but not in radish and carnation plants
(Van Peer et al. 1991; Van Peer and Schippers 1992; Leeman et al. 1995; Van Wees
et al. 1997).
Antibiotics are one or more compounds secreted by one microbial species at low
concentrations to inhibit/kill other microbial species. BCAs must produce antibiot-
ics biosynthesized by different genes in sufficient quantities near the pathogen to
induce positive biological control. Several BCAs that produce in situ antibiotics
have been measured (Thomashow et al. 2002); however, the effective quantities are
still required to be estimated because of the presence of other organic compounds.
Few examples of BCAs that are known to produce multiple antibiotics are Bacillus
cereus strain UW85 that produces both zwittermicin (Silo-Suh et al. 1994) and
kanosamine and can suppress one or more pathogens. This ability of producing dif-
ferent antibiotics by single bacterial species suppresses diverse plant pathogens and
thus enhances biological control in rice and maize cropping systems.
References
Agnihotri S, Kulshreshtha K, Singh SN (1999) Mitigation strategy to contain methane emission
from rice-fields. Environ Monit Assess 58:95–104
Ahmad M, Nadeem SM, Naveed M, Zahir ZA (2016) Potassium-solubilizing bacteria and their
application in agriculture. In: Meena VS, Maurya BR, Verma JP, Meena RS (eds) Potassium
solubilizing microorganisms for sustainable agriculture. Springer, New Delhi, pp 293–313.
doi:10.1007/978-81-322-2776-2_21
Alabouvette C (1999) Fusarium wilt suppressive soils: an example of disease suppressive soils.
Aust Plant Pathol 28:57–64
Ali MA, Farouque G, Haque M, Kabir A (2012) Influence of soil amendments on mitigating
methane emissions and sustaining rice productivity in paddy soil ecosystems of Bangladesh.
J Environ Sci Nat Resour 5:179–185
Audenaert K, Pattery T, Cornelis P, Hofte M (2002) Induction of systemic resistance to Botrytis
cinerea in tomato by Pseudomonas aeruginosa 7NSK2: role of salicylic acid, pyochelin, and
pyocyanin. Mol Plant Microbe Int 15:1147–1156
Azcona I, Pascual I, Aguirreolea J, Fuentes M, Garcia-Mina JM, Sanchez-Diaz M (2011) Growth
and development of pepper are affected by humic substances derived from composted sludge.
J Plant Nutr Soil Sci 174:916–924
Bahadur I, Meena VS, Kumar S (2014) Importance and application of potassic biofertilizer in
Indian agriculture. Int Res J Biol Sci 3:80–85
Bahadur I, Maurya BR, Kumar A, Meena VS, Raghuwanshi R (2016a) Towards the soil sustain-
ability and potassium-solubilizing microorganisms. In: Meena VS, Maurya BR, Verma JP,
Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture. Springer,
New Delhi, pp 225–266. doi:10.1007/978-81-322-2776-2_18
Bahadur I, Maurya BR, Meena VS, Saha M, Kumar A, Aeron A (2016b) Mineral release dynamics
of tricalcium phosphate and waste muscovite by mineral-solubilizing rhizobacteria isolated
from indo-gangetic plain of India. Geomicrobiol J. doi:10.1080/01490451.2016.1219431
Bakker PAHM, Pieterse CMJ, Van Loon LC (2007) Induced systemic resistance by fluorescent
Pseudomonas spp. Phytopathology 97:239–243
Barea JM (2015) Future challenges and perspectives for applying microbial biotechnology in sus-
tainable agriculture based on a better understanding of plant-microbiome interactions. J Soil
Sci Plant Nutr 15:261–282
Barea JM, Richardson AE (2015) Phosphate mobilisation by soil microorganisms. In: Lugtenberg
B (ed) Principles of plant-microbe interactions. Springer, Heidelberg, pp 225–234
Barea JM, Pozo MJ, Azcon R, Azcon-Aguilar C (2013) Microbial interactions in the rhizosphere.
In: de Bruijn FJ (ed) Molecular microbial ecology of the rhizosphere, vol 1. Wiley Blackwell,
Hoboken, pp 29–44
Browne P, Barret M, Morrissey JP, O’Gara F (2013) Molecular-based strategies to exploit the inor-
ganic phosphate-solubilization ability of Pseudomonas in sustainable agriculture. In: de Bruijn
FJ (ed) Molecular microbial ecology of the rhizosphere, vol 2. Wiley Blackwell, Hoboken,
pp 615–628
de Bruijn FJ (2015) Biological nitrogen fixation. In: Lugtenberg B (ed) Principles of plant-microbe
interactions. Springer, Heidelberg, pp 215–224
Compant S, Duffy B, Nowak J, Clement C, Barka EA (2005) Use of plant growth-promoting bac-
teria for biocontrol of plant diseases: principles, mechanisms of action, and future prospects.
Appl Environ Microbiol 71:4951–4959
Da Rocha AB, Hammerschmidt R (2005) History and perspectives on the use of disease resistance
inducers in horticultural crops. Hortic Technol 15:518–529
Das I, Pradhan M (2016) Potassium-solubilizing microorganisms and their role in enhancing
soil fertility and health. In: Meena VS, Maurya BR, Verma JP, Meena RS (eds) Potassium
solubilizing microorganisms for sustainable agriculture. Springer, New Delhi, pp 281–291.
doi:10.1007/978-81-322-2776-2_20
13 Efficacy of Biological Soil Amendments and Biocontrol Agents for Sustainable… 293
Kumar A, Bahadur I, Maurya BR, Raghuwanshi R, Meena VS, Singh DK, Dixit J (2015) Does
a plant growth-promoting rhizobacteria enhance agricultural sustainability? J Pure Appl
Microbiol 9:715–724
Kumar A, Meena R, Meena VS, Bisht JK, Pattanayak A (2016a) Towards the stress management
and environmental sustainability. J Clean Prod 137:821–822
Kumar A, Patel JS, Bahadur I, Meena VS (2016b) The molecular mechanisms of KSMs for
enhancement of crop production under organic farming. In: Meena VS, Maurya BR, Verma JP,
Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture. Springer,
New Delhi, pp 61–75. doi:10.1007/978-81-322-2776-2_5
Kumar A, Maurya BR, Raghuwanshi R, Meena VS, Islam MT (2017) Co-inoculation with
Enterobacter and Rhizobacteria on yield and nutrient uptake by wheat (Triticum aestivum L.)
in the alluvial soil under indo-gangetic plain of India. J Plant Growth Regul. doi:10.1007/
s00344-016-9663-5
Kyselkova M, Kopecky J, Frapolli M, Defago G, Sagova-Mareckova M, Grundmann GL, Moenne-
Loccoz Y (2009) Comparison of rhizobacterial community composition in soil suppressive or
conducive to tobacco black root rot disease. ISME J 3:1127–1138
Lee YB, Lee CH, Hawng JY, Lee IB, Kim PJ (2004) Enhancement of phosphate desorption by
silicate in soils with salt accumulation. Soil Sci Plant Nutr 50:43–499
Leeman M, Van Pelt JA, Den Ouden FM, Heinsbroek M, Bakker PAHM, Schippers B (1995)
Induction of systemic resistance against fusarium wilt of radish by lipopolysaccharides of
Pseudomonas fluorescens. Phytopathology 85:1021–1027
Leeman M, den Ouden EM, van Pelt JA, Dirkx FPM, Steijl H, Bakker PAHM, Schippers B
(1996) Iron availability affects induction of systemic resistance to Fusarium wilt of radish by
Pseudomonas fluorescens. Phytopathology 86:149–155
Lugtenberg B (2015) Life of microbes in the rhizosphere. In: Lugtenberg B (ed) Principles of
plant-microbe interactions. Springer, Heidelberg, pp 7–15
Ma BL, Dwyer LM, Gregorich ED (1999) Soil N amendment effects on seasonal N mineralization
and N cycling in maize production. Agron J 91:1003–1009
Manandhar HK, Lyngs Jorgensen HJ, Mathur SB, Smedegaard-Peterson V (1998) Suppression
of rice blast by preinoculation with avirulent Pyricularia oryzae and the non-rice pathogen
Bipolaris sorokiniana. Phytopathology 88:735–739
Masood S, Bano A (2016) Mechanism of potassium solubilization in the agricultural soils by the
help of soil microorganisms. In: Meena VS, Maurya BR, Verma JP, Meena RS (eds) Potassium
solubilizing microorganisms for sustainable agriculture. Springer, New Delhi, pp 137–147.
doi:10.1007/978-81-322-2776-2_10
Maurya BR, Meena VS, Meena OP (2014) Influence of inceptisol and alfisol’s potassium solubiliz-
ing bacteria (KSB) isolates on release of K from waste mica. Vegetos 27:181–187
Mazzola M (2002) Mechanisms of natural soil suppressiveness to soilborne diseases. Anton
Leeuw Int J G 81:557–564
Meena OP, Maurya BR, Meena VS (2013a) Influence of K-solubilizing bacteria on release of
potassium from waste mica. Agric Sustain Dev 1:53–56
Meena VS, Maurya BR, Bohra JS, Verma R, Meena MD (2013b) Effect of concentrate manure
and nutrient levels on enzymatic activities and microbial population under submerged rice in
alluvium soil of Varanasi. Crop Res 45(1,2 & 3):6–12
Meena VS, Maurya BR, Verma R, Meena RS, Jatav GK, Meena SK, Meena SK (2013c) Soil
microbial population and selected enzyme activities as influenced by concentrate manure and
inorganic fertilizer in alluvium soil of Varanasi. Bioscan 8(3):931–935
Meena VS, Maurya BR, Bahadur I (2014a) Potassium solubilization by bacterial strain in waste
mica. Bangladesh J Bot 43:235–237
Meena VS, Maurya BR, Verma JP (2014b) Does a rhizospheric microorganism enhance K+ avail-
ability in agricultural soils? Microbiol Res 169:337–347
Meena RS, Meena VS, Meena SK, Verma JP (2015a) The needs of healthy soils for a healthy
world. J Clean Prod 102:560–561
13 Efficacy of Biological Soil Amendments and Biocontrol Agents for Sustainable… 295
Meena RS, Meena VS, Meena SK, Verma JP (2015b) Towards the plant stress mitigate the agricul-
tural productivity: a book review. J Clean Prod 102:552–553
Meena VS, Maurya BR, Meena RS (2015c) Residual impact of well grow formulation and NPK on
growth and yield of wheat (Triticum aestivum L.) Bangladesh J Bot 44(1):143–146
Meena VS, Maurya BR, Verma JP, Aeron A, Kumar A, Kim K, Bajpai VK (2015d) Potassium
solubilizing rhizobacteria (KSR): isolation, identification, and K-release dynamics from waste
mica. Ecol Eng 81:340–347
Meena VS, Meena SK, Verma JP, Meena RS, Ghosh BN (2015e) The needs of nutrient use efficiency
for sustainable agriculture. J Clean Prod 102:562–563. doi:10.1016/j.jclepro.2015.04.044
Meena VS, Verma JP, Meena SK (2015f) Towards the current scenario of nutrient use efficiency in
crop species. J Clean Prod 102:556–557. doi:10.1016/j.jclepro.2015.04.030
Meena RK, Singh RK, Singh NP, Meena SK, Meena VS (2016a) Isolation of low temperature sur-
viving plant growth-promoting rhizobacteria (PGPR) from pea (Pisum sativum L.) and docu-
mentation of their plant growth promoting traits. Biocatal Agric Biotechnol 4:806–811
Meena RS, Bohra JS, Singh SP, Meena VS, Verma JP, Verma SK, Sihag SK (2016b) Towards the
prime response of manure to enhance nutrient use efficiency and soil sustainability a current
need: a book review. J Clean Prod 112(1):1258–1260
Meena SK, Rakshit A, Meena VS (2016c) Effect of seed bio-priming and N doses under varied
soil type on nitrogen use efficiency (NUE) of wheat (Triticum aestivum L.) under greenhouse
conditions. Biocatal Agric Biotechnol 6:68–75
Meena VS, Bahadur I, Maurya BR, Kumar A, Meena RK, Meena SK, Verma JP (2016d) Potassium-
solubilizing microorganism in evergreen agriculture: an overview. In: Meena VS, Maurya BR,
Verma JP, Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture.
Springer, New Delhi, pp 1–20. doi:10.1007/978-81-322-2776-2_1
Meena VS, Meena SK, Bisht JK, Pattanayak A (2016e) Conservation agricultural practices in
sustainable food production. J Clean Prod 137:690–691
Mendes R, Kruijt M, De Bruijn I, Dekkers E, Van der Voort M, Schneider JHM, Piceno YM,
DeSantis TZ, Andersen GL, Bakker PAHM, Raaijmakers JM (2011) Deciphering the rhizo-
sphere microbiome for disease-suppressive bacteria. Science 332:1097–1100
Mercado-Blanco J (2015) Life of microbes inside the plant. In: Lugtenberg B (ed) Principles of
plant-microbe interactions. Springer, Heidelberg, pp 25–32
Meziane H, Van der Sluis I, Van Loon LC, Hofte M, Bakker PAHM (2005) Determinants of
Pseudomonas putida WCS358 involved in inducing systemic resistance in plants. Mol Plant
Pathol 6:177–185
Mishina TE, Zeier J (2007) Pathogen-associated molecular pattern recognition rather than devel-
opment of tissue necrosis contributes to bacterial induction of systemic acquired resistance in
Arabidopsis. Plant J 50:500–513
Nardi S, Carletti P, Pizzeghello D, Muscolo A (2009) Biological activities of humic substances. In:
Senesi N, Xing B, Huang PM (eds) Biophysico-chemical processes involving natural nonliv-
ing organic matter in environmental systems, PART I. Fundamentals and impact of mineral-
organic-biota interactions on the formation, transformation, turnover, and storage of natural
nonliving organic matter (NOM). Wiley, Hoboken, pp 301–335
Olivares J, Bedmar EJ, Sanjuan J (2013) Biological nitrogen fixation in the context of global
change. Mol Plant-Microbe Interact 26:486–494
Ongena M, Jourdan E, Schafer M, Kech C, Budzikiewicz H, Luxen A, Thonart P (2005) Isolation
of an N-alkylated benzylamine derivative from Pseudomonas putida BTP1 as elicitor of
induced systemic resistance in bean. Mol Plant-Microbe Interact 18:562–569
Ongena M, Jourdan E, Adam A, Paquot M, Brans A, Joris B, Arpigny JL, Thonart P (2007)
Surfactin and fengycin lipopeptides of Bacillus subtilis as elicitors of induced systemic resis-
tance in plants. Environ Microbiol 9:1084–1090
Ordentlich A, Elad Y, Chet I (1988) The role of chitinase of Serratia marcescens in biocontrol of
Sclerotium rolfsii. Phytopathology 78:84–88
Parewa HP, Yadav J, Rakshit A, Meena VS, Karthikeyan N (2014) Plant growth promoting rhizo-
bacteria enhance growth and nutrient uptake of crops. Agric Sustain Dev 2(2):101–116
296 M. Kaushal and S.P. Wani
Pozo MJ, Verhage A, Garcia-Andrade J, Garcia JM, Azcon-Aguilar C (2009) Priming plant
defences against pathogens by arbuscular mycorrhizal fungi. In: Azcon-Aguilar C, Barea JM,
Gianinazzi S, Gianinazzi-Pearson V (eds) Mycorrhizas: functional processes and ecological
impact. Springer, Heidelberg, pp 137–149
Prakash S, Verma JP (2016) Global perspective of potash for fertilizer production. In: Meena VS,
Maurya BR, Verma JP, Meena RS (eds) Potassium solubilizing microorganisms for sustainable
agriculture. Springer, New Delhi, pp 327–331. doi:10.1007/978-81-322-2776-2_23
Press CM, Wilson M, Tuzun S, Kloepper JW (1997) Salicylic acid produced by Serratia marc-
escens 90–166 is not the primary determinant of induced systemic resistance in cucumber or
tobacco. Mol Plant Microbe Int 10:761–768
Priyadharsini P, Muthukumar T (2016) Interactions between arbuscular mycorrhizal fungi and
potassium-solubilizing microorganisms on agricultural productivity. In: Meena VS, Maurya
BR, Verma JP, Meena RS (eds) Potassium solubilizing microorganisms for sustainable agricul-
ture. Springer, New Delhi, pp 111–125. doi:10.1007/978-81-322-2776-2_8
Raaijmakers JM, Lugtenberg BJJ (2013) Perspectives for rhizosphere research. In: de Bruijn
FJ (ed) Molecular microbial ecology of the rhizosphere, vol 2. Wiley Blackwell, Hoboken,
pp 1227–1232
Raaijmakers JM, Weller DM (1998) Natural plant protection by 2,4-diacetylphloroglucinol-
producing Pseudomonas spp. in take-all decline soils. Mol Plant-Microbe Interact 11:144–152
Raghavendra MP, Nayaka NC, Nuthan BR (2016) Role of rhizosphere microflora in potas-
sium solubilization. In: Meena VS, Maurya BR, Verma JP, Meena RS (eds) Potassium
solubilizing microorganisms for sustainable agriculture. Springer, New Delhi, pp 43–59.
doi:10.1007/978-81-322-2776-2_4
Raimam MP, Albino U, Cruz MF, Lovato GM, Spago F, Ferracin TP, Lima DS, Goulart T, Bernardi
CM, Miyauchi M, Nogueira MA, Andrade G (2007) Interaction among free-living N-fixing
bacteria isolated from Drosera villosa var. villosa and AM fungi (Glomus clarum) in rice
(Oryza sativa). Appl Soil Ecol 35:25–34
Ran LX, Li ZN, Wu GJ, Van Loon LC, Bakker PAHM (2005) Induction of systemic resistance
against bacterial wilt in Eucalyptus urophylla by fluorescent Pseudomonas spp. Eur J Plant
Pathol 113:59–70
Rawat J, Sanwal P, Saxena J (2016) Potassium and its role in sustainable agriculture. In: Meena
VS, Maurya BR, Verma JP, Meena RS (eds) Potassium solubilizing microorganisms for sus-
tainable agriculture. Springer, New Delhi, pp 235–253. doi:10.1007/978-81-322-2776-2_17
Reglinski T, Walters D (2009) Induced resistance for plant disease control. In: Walters D (ed)
Disease control in crops. Wiley-Blackwell, Oxford, pp 62–92
Reid LM, Zhu X, Ma BL (2001) Crop rotation and nitrogen effects on maize susceptibility to gib-
berella (Fusarium graminearum) ear rot. Plant Soil 237:1–14
Ryu CM, Farag MA, Hu CH, Reddy MS, Kloepper JW, Pare PW (2004) Bacterial volatiles induce
systemic resistance in Arabidopsis. Plant Physiol 134:1017–1026
Saha M, Maurya BR, Bahadur I, Kumar A, Meena VS (2016a) Can potassium-solubilising bacteria
mitigate the potassium problems in India? In: Meena VS, Maurya BR, Verma JP, Meena RS
(eds) Potassium solubilizing microorganisms for sustainable agriculture. Springer, New Delhi,
pp 127–136. doi:10.1007/978-81-322-2776-2_9
Saha M, Maurya BR, Meena VS, Bahadur I, Kumar A (2016b) Identification and characterization
of potassium solubilizing bacteria (KSB) from Indo-Gangetic Plains of India. Biocatal Agric
Biotechnol 7:202–209
Sanguin H, Sarniguet A, Gazengel K, Moenne-Loccoz Y, Grundmann GL (2009) Rhizosphere bac-
terial communities associated with disease suppressiveness stages of take-all decline in wheat
monoculture. New Phytol 184:694–707
Schreiner K, Hagn A, Kyselkova M, Moenne-Loccoz Y, Welzl G, Munch JC, Schloter M (2010)
Comparison of barley succession and take-all disease as environmental factors shaping the
rhizobacterial community during take-all decline. Appl Environ Microbiol 76:4703–4712
Schuhegger R, Ihring A, Gantner S, Bahnweg G, Knappe C, Vogg G, Hutzler P, Schmid M, Van
Breusegem F, Eberl L, Hartmann A, Langebartels C (2006) Induction of systemic resistance
13 Efficacy of Biological Soil Amendments and Biocontrol Agents for Sustainable… 297
Abstract
The increasing world population has forced the world to improve the productivity
of crops. Among other important cereals, wheat is one such a crop which has
increasing demands all over the world. It provides ~20% of caloric and protein
needs worldwide. Although the use of modern technologies enhanced the
production of wheat, it also resulted in decreased mineral content of crops, which
include Ca, K, Na, Zn, Fe and Se. The selenium (Se) content in wheat is highly
variable ranging from 5 to 720 μg Se/kg. Meanwhile the Se distribution is highly
uneven worldwide, ranging from a low value of 0.01 mg Se/kg of soil to a higher
value of 1200 mg Se/kg of soil in seleniferous soils. Wheat is among one of the
major sources of Se supply for humans in areas where wheat is an important
component of daily diet, so it must be maintained in healthy concentration to
ensure adequate supply of Se to humans. The use of selenate and selenite with
urea crystals and foliar application proved to enhance selenium content, but these
measures have their own drawbacks. Bacterial inoculation to wheat resulted in
remarkable increase in the accumulation of selenium in wheat plant along with
Ni, Mn, P, Mo, K, Fe and Ca. Selenium-fortified wheat is a best option for the
daily intake of selenium by humans.
Keywords
Biofortification • Phytoremediation • Wheat • Selenium • Bacteria
14.1 Introduction
The selenium (Se) is a metalloid, which belongs to the group VI-A of the periodic
table. Selenium shows several beneficial as well as toxic effects on organisms that
depend on the quantity of available Se in soil for plant uptake, accumulation in food
stuff and its daily intake by humans and other organisms. Selenium requirement for
humans is ~55 to 60 μg day−1; however, daily intake of <40 μg Se may lead to Se
deficiency, and daily intake of ~400 μg Se may cause toxic effects in humans (WHO
1996). Se exists in six isotopes with 80Se as the most abundant one. In nature, Se is
found in four different oxidation states, 2 in halides, 0 in elemental form, 4+in sel-
enite and 6+in selenate. It belongs to group VI-A, and its chemical properties are
very similar to sulphur (S). In environment, Se occurs in both inorganic and organic
forms in soil, water and air. Inorganic forms include selenite/selenate salts, while
organic forms are the Se products from decomposition of Se-rich plant material
(Barceloux and Barceloux 1999). Shale rock-derived soils are one of the natural rich
sources of Se. Selenium distribution is highly uneven worldwide, and Se quantity in
soil may vary from a low value of 0.01 to 2 mg Se kg−1 to a high value of 1200 mg
Se kg−1 of soil (Mayland 1994). Anthropogenic activities are one of the major fac-
tors for the imbalance of Se in the environment, e.g. the use of Se metalloid in
metallurgy and glass manufacturing industry constitutes ~60% of the total Se used
(USGS 2011).
Selenium plays a significant role in the biological systems, as it is the main com-
ponent of many selenoproteins in living organisms. About 30 selenoproteins are
encoded by 25 human genes. Glutathione peroxidases (GPxs) are such eight pro-
teins (GPx1-GPx8) having antioxidant properties with multidimensional roles in
living cells, ranging from H2O2 homeostasis to regulation of apoptosis (Brigelius-
Flohe and Maiorino 2013). Other selenoproteins are deiodinases and thioredoxin
reductases which play a significant role in thyroid hormone metabolism, intracel-
lular redox potential, DNA synthesis, cancer prevention, fertility and reproduction
(Suttle 2010). Selenium in combination with vitamin E plays a synergistic effect on
muscle activity as it improves endurance and recovery of muscles cells (Cabaraux
et al. 2007).
Low Se content in diet may reduce the efficiency of immune system which may
result in occurrence of various diseases, both in animals and humans (Table 14.1).
In animals, deficiency of Se affects the heart, skeletal muscles and liver. White mus-
cle disease, a degenerative myopathy, is a common disease of animals due to Se
deficiency. Commonly cattle, poultry and horses are affected by this disease. In
humans Se deficiency results in Keshan disease (cardiomyopathy) and Kashin-Beck
disease (osteoarthropathy). Investigations showed that these diseases are common
in areas where daily intakes are between 10 and 15 μg Se day−1.
On the other hand, high Se intake may result in low haemoglobin level, decreased
sperm motility and increased incidence of pancreatic and skin cancer. It is estimated
that over 800 million people worldwide might have been affected due to selenium
deficiency (Fordyce 2013; Meena et al. 2013a, 2016a; Bahadur et al. 2014; Maurya
et al. 2014; Jat et al. 2015; Kumar et al. 2015, 2016b; Ahmad et al. 2016).
14 Bacterial-Mediated Selenium Biofortification of Triticum aestivum: Strategy… 301
14.2 W
orldwide Importance of Wheat (Triticum aestivum)
as Food Crop
Wheat is one of the oldest forms of nutrition available to humans and considered to
be one of the main crops to ensure global food security. Introduction of new tech-
nologies in agriculture sector and development of high-yield strains of crops had
helped a lot to reduce hunger and poverty worldwide. In developing countries, it is
302 Z.u. Abadin et al.
Table 14.2 Bacterial-associated Se biofortification in different crops and its effects on Se uptake
in plants
Mineral for
Crop biofortification Bacteria used Results References
Wheat Selenium Paenibacillus sp.; Bacillus Se associated Acuña et al.
sp. with STB was (2013)
translocated into
the leaves
Wheat Selenium, iron Bacillus sp. Increase in Se and Yasin et al.
Fe amount in (2015a)
wheat kernels
Enhanced growth
of plants
Wheat Selenium Bacillus sp., Paenibacillus P-solubilization, Duran et al.
sp., Klebsiella, high IAA (2014)
Acinetobacter sp. production
Wheat Iron, copper, Providencia sp., Increased protein Rana et al.
zinc Anabaena sp., Calothrix content (2012)
sp.
Saltmarsh Selenium, Rhizosphere bacteria Se and Hg were De Souza
bulrush, mercury accumulated in et al.
rabbitfoot wetland plants (1999)
grass
Wheat Selenium Stenotrophomonas sp., Rise in selenium Duran et al.
Enterobacter sp., Bacillus content of wheat (2013)
sp., Pseudomonas sp., G. grains
claroideum
Chickpea Iron PGPR Increase in iron Khalid
content of grain et al.
and shoot (2015)
projected that the demand will increase by ~60% till 2050 (Nelson et al. 2010). To
meet this increasing demand, several developing countries are importing colossal
amounts of wheat, and it is estimated that wheat has the highest percentage (~46%)
out of total food imports (Dixon et al. 2009).
Wheat is an important source of calories and proteins. It helps to meet one fifth
of daily calories and proteins to human population. According to the available data,
~19% of daily caloric needs and ~21% of daily protein intake globally are met by
wheat consumption. Developing countries have higher demand for wheat. In Central
Asia ~47% of the daily calories and proteins are provided by wheat intake; on the
other hand this percentage falls dramatically to ~20% in high-income countries
(Parewa et al. 2014; Prakash and Verma 2016; Meena et al. 2015f, 2016b;
Priyadharsini and Muthukumar 2016; Jaiswal et al. 2016; Jha and Subramanian
2016; Kumar et al. 2016a).
However, the ~2.5 billion deprived people of the world are dependent on wheat
as the main component of their diet. With regard to wheat as an important cereal
crop worldwide, Se-supplemented or Se-rich wheat grains can be an important
14 Bacterial-Mediated Selenium Biofortification of Triticum aestivum: Strategy… 303
14.3 C
ontent of Se in Wheat Under Natural Conditions
Worldwide
Selenium content of the soil varies significantly across the globe, which in turn
affects the Se content of crops growing in those regions. Studies show that selenium
content of wheat varies from as low as 0.001 mg/kg in the areas of southwestern
Australia to as high as 30 mg/kg in some areas of South Dakota, whereas the world’s
average wheat content is between the range of 0.02 and 0.60 mg Sekg−1. A study
conducted by Lyons et al. (2005a) in different areas of Mexico and Australia showed
the variation of the Se content of wheat varieties in these regions. The results of the
study provided a range 5–720 μg Se kg−1 in wheat samples. Survey conducted to
find out Se content of wheat grown at Centro Internacional de Mejoramiento de
Maiz y Trigo (International Maize and Wheat Improvement Centre) showed a range
from 9 to 244 μg Se kg−1 in hundred wheat samples, whereas at the same place, field
trials showed a range of 40–110 μg Se kg−1.
Field trials of wheat grown at three different soils near Tamworth, New South
Wales, over a period of 27 years also showed large variations in Se content. Neutral
soils showed large amount of Se in wheat ranging from 61 to 510 μg Sekg−1 in 27
samples of wheat during the year of 1971. This range narrowed in 1996 and became
140–490 μg Sekg−1. While for black cracking soil with slightly acidic pH, the range
almost remained the same, i.e. from 35 to 210 μg Se kg−1, over the period. For more
acidic soil, Se content of wheat was very low it ranged from 18 to 22 μg Se kg−1.
These results also supported the idea that Se absorption by plants decreases in acidic
soils (Lyons et al. 2005a). Selenium concentrations determined in different areas of
the world are shown in Table 14.4.
304 Z.u. Abadin et al.
There are many factors which determine availability of Se in soil (Fig. 14.1). These
factors include organic matter content, pH of soil, redox conditions in the crop field,
speciation of Se, soil texture and mineralogy, etc. (Fordyce 2013).
In natural conditions, the inorganic form of Se, i.e. selenite (SeO3−2), is more
stable because it is absorbed by ligand exchange which is pH dependent. Decrease
in pH increases adsorption by ligand exchange. This adsorption and formation of
insoluble iron oxide and oxyhydroxide makes selenite less available for absorption
by plants (Fig. 14.1). On the other hand, selenate (SeO4−2) is more soluble and
mobile at neutral and alkaline pH, that is why it is easily taken up by plants. This is
also supported by comparing uptake of selenate and selenite by plants (Alloway
2013; Kumar et al. 2017; Meena et al. 2015a, b; Raghavendra et al. 2016; Zahedi
2016; Rawat et al. 2016; Yasin et al. 2016; Das and Pradhan 2016; Dominguez-
Nunez et al. 2016; Dotaniya et al. 2016).
Other inorganic forms of Se present in soil are selenides (Se2−) and elemental
selenium (Se0), which are present in acidic and organic-rich environment. These
forms of Se are less available to plants due to their low solubility and oxidation
potential. Organic forms of selenium in soil are selenomethionine (C5H11NO2Se)
and selenocysteine (C3H7NO2Se). Both of these organic forms are readily available
to plants. Order of Se bioavailability of these forms of Se, starting with the most
soluble and mobile forms are as follows: selenate (Se+6) > selenomethionine > sele-
nocysteine > selenite (Se+4) > selenium (Se0) > selenide (Se−2) (Jacobs 1989; Neal
1995; Yanai et al. 2015).
14 Bacterial-Mediated Selenium Biofortification of Triticum aestivum: Strategy… 305
Fig. 14.1 Factors
influencing Se availability Less Bioavailability Higher Bioavailability
in soil (Fordyce 2013)
pH < 7 pH > 7
Reducing oxidizing
conditions conditions
Immobile mobile
SeO32+ SeO 2+
4
SeO 2+
4
14.6 R
ole of Selenobacteria (Se-Tolerant Bacteria) in Wheat
Growth Promotion and Selenium Uptake
Numerous studies (Yasin et al. 2015a; Duran et al. 2014) in the last two decades
support the idea of microbe-supplemented biofortification of wheat. Selenobacteria
(Se-tolerant bacteria) isolated from different ecological niches have been explored
and studied for their potential in the bioremediation and biofortification of soils
(Eswayah et al. 2016). Duran et al. (2013) showed that inoculation of bacteria
enhanced translocation of Se to shoots and ultimately increased the Se content of
the wheat grain. Another study conducted by same research group shows that ACC
deaminase activity of selenobacteria not only protects plant against stress but also
improves the Se content of plant and eliminates stress produced due to Se toxicity
(Acuna et al. 2013).
Studies conducted by Yasin et al. (2015a) on wheat also showed strong relation
between selenobacteria (Fig. 14.2) and wheat growth and their positive role in bio-
fortification of Se and other essential mineral elements in plants. Bacillus strains
with high tolerance for Se showed significantly improved Se uptake in wheat plants
and different plant growth-enhancing attributes including auxin production, more
differentiated roots, greater number of leaves, greater dry weight and increased
shoot length and higher acid phosphatase activity (Yasin et al. 2015b). Another
study by our group showed that bacterial inoculation increased acid phosphatase
activity in T. aestivum by ~40% (Yasin and Faisal 2013).
14 Bacterial-Mediated Selenium Biofortification of Triticum aestivum: Strategy… 307
Less work was carried out on the transport of selenate and selenite into the bacterial
cells. Available data showed that two different mechanisms are used by bacteria for
uptake of these oxyanions. Selenate ions are transported by sulphate ATP-binding
cassette transporter complex (consists of five proteins) which is encoded by the
cysAWTP operon (Sirko et al. 1990). This complex includes two CysA, which are
ATP-binding proteins; CysT and CysW, which are transmembrane proteins; and
CysP which is periplasmic sulphate-binding protein. While selenite is transported
by sulphate permease in E. coli along with ABC transporter complex, blocking of
transport complex does not completely prevent the selenite uptake (Turner et al.
1998; Rosen and Liu 2009).
As discussed earlier, Se has similar properties as that of S, so they have some
common enzymes for their metabolism. Selenium non-accumulator plants cannot
tolerate high Se, and they contain generally less than 25 μg Se gramme−1 dry weight
(DW) of the plant (White et al. 2004), whereas hyperaccumulator species of plants
have high selenium content of 10–15 mg Se gramme−1dry weight of plant. One
hypothesis is Se hyperaccumulator plants have the ability to bear high Se content
due to their capability to restrict integration of selenomethionine and selenocysteine
(SeCys) into cellular proteins (Brown and Shrift 1982; El Mehdawi and Pilon-Smits
2012). This restriction is achieved by conversion of SeMet into amino acids which
are not involved in protein formation like ɤ-glutamyl-Se-methylselenocysteine,
Se-methylselenocysteine and selenocystathionine (Burnell 1981). The available
data shows that Se is not required by plants as there is no codon found for SeCys in
plants. However, plants are ought to be significant sources of Se for animal and
humans (Sors et al. 2005a, b).
308 Z.u. Abadin et al.
Three readily available forms of Se in soil are selenate, selenite and organic com-
pounds of Se. Uptake of these compounds depends on physiochemical status of soil,
i.e. amount of phosphate and sulphate ions and pH. Selenate and sulphate ions com-
pete for transport into the cell; however, these ions are transported against their
electrochemical gradients, by cotransport of three protons against one ion (Boldrin
et al. 2016). Sulphate is transported via high- and low-affinity sulphate transporter
proteins. Literature review showed that high-affinity sulphate transporters are
responsible for selenate transport into the plant (Shibagaki et al. 2002). On the other
hand, selenite and organic Se compounds have different mechanisms for their trans-
port into the cell. There uptake is passive as compared to selenate which is taken
actively (Abrams et al. 1990; Supriatin et al. 2016).
Experiments with wheat showed similar results for the transport and assimilation
of Se, and selenate uptake was through high-affinity sulphate transporters in wheat,
because S starvation increased selenate uptake. Similarly, selenite uptake in wheat
roots is through phosphate transporters, and it is metabolically dependent active
process. Assimilation of these two oxyanions is also different as selenate remains in
roots and shoots, especially in xylem sap, because its assimilation into organic com-
pounds is quite low, whereas selenite is rapidly metabolized into organic forms in
plant tissues. Furthermore, studies show that selenite presence in the soil inhibits
selenate uptake (Li et al. 2008; Verma et al. 2014, 2015a, b; Meena et al. 2013b,
2014a, 2016e; Sharma et al. 2016; Masood and Bano 2016; Teotia et al. 2016).
14.8 S
elenium Fortification Strategies and Their Worldwide
Application
There are various methods and strategies to improve Se content in food and its con-
sumptions by humans. One way is to take Se directly either by supplementation of
livestock or by Se pills, while the other way, as discussed in this chapter, is to use
agronomic biofortification strategies. Agronomic biofortification is an important
tool to enhance Se status of crops. Several recent studies proved that this strategy
enhances Se content of crops (Ros et al. 2016). Another strategy is to apply Se fertil-
izers directly to the soil; however, Se uptake in plants may be affected by various
factors such as application time of specific fertilizer, chemical speciation and type
of Se in soil. Among the inorganic anions such as selenate and selenite, supply of
selenate to plants results in the highest crop response regarding Se uptake in plants
(Cartes et al. 2005).
Another strategy is the foliar application of Se fertilizer. This strategy is more
effective as compared to Se application through soil, as studies showed that it almost
doubles the Se content of the crop. A study by Ducsay and Lozek (2006) showed
that the foliar application of 10–20 g Se hector−1 increased the Se content of wheat
grains. Se content in unfertilized samples was 0.045 mg/kg−1dry weight, whereas
when 10 g Sehector−1 was applied, this content was raised to 0.088 mg/kg dry
weight. It was a combination of foliar and soil-applied Se because the foliar fertil-
izers are applied before extensive leaf cover. Such fertilizers showed different crop
14 Bacterial-Mediated Selenium Biofortification of Triticum aestivum: Strategy… 309
14.9 Conclusions
Acknowledgements The author would like to thank Higher Education Commission Pakistan and
University of the Punjab, Lahore, Pakistan, for the support for the research work to our group.
References
Abrams M, Burau R, Zasoski R (1990) Organic selenium distribution in selected California soils.
Soil Sci Soc Am J 54(4):979–982
Acuña JJ, Jorquera MA, Barra PJ, Crowley DE, Mora ML (2013) Selenobacteria selected from
the rhizosphere as a potential tool for se biofortification of wheat crops. Biol Fert Soils
49(2):175–185
Ahmad M, Nadeem SM, Naveed M, Zahir ZA (2016) Potassium-solubilizing bacteria and their
application in agriculture. In: Meena VS, Maurya BR, Verma JP, Meena RS (eds) Potassium
solubilizing microorganisms for sustainable agriculture. Springer, India, pp 293–313.
doi:10.1007/978-81-322-2776-2_21
Alloway BJ (2013) Introduction. Heavy metals in soils. Springer, Dordrecht, pp 3–9
Bahadur I, Meena VS, Kumar S (2014) Importance and application of potassic biofertilizer in
Indian agriculture. Int Res J Biol Sci 3:80–85
310 Z.u. Abadin et al.
Bahadur I, Maurya BR, Kumar A, Meena VS, Raghuwanshi R (2016a) Towards the soil sustain-
ability and potassium-solubilizing microorganisms. In: Meena VS, Maurya BR, Verma JP,
Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture. Springer,
India, pp 225–266. doi:10.1007/978-81-322-2776-2_18
Bahadur I, Maurya BR, Meena VS, Saha M, Kumar A, Aeron A (2016b) Mineral release dynamics
of tricalcium phosphate and waste muscovite by mineral-solubilizing rhizobacteria isolated
from indo-gangetic plain of India. Geomicrobiol J. doi:10.1080/01490451.2016.1219431
Banerjee MR, Yesmin L (2009) Sulfur-oxidizing plant growth promoting rhizobacteria for
enhanced canola performance, Google Patents number US7491535
Barceloux DG, Barceloux D (1999) Selenium. J Toxicol Clin Toxicol 37(2):145–172
Berendsen RL, Pieterse CM, Bakker PA (2012) The rhizosphere microbiome and plant health.
Trends Plant Sci 17(8):478–486
Berg G (2009) Plant–microbe interactions promoting plant growth and health: perspectives for
controlled use of microorganisms in agriculture. Appl Microbiol Biotechnol 84(1):11–18. Biol
Fert Soil 50(6): 983–990
Boldrin PF, de Figueiredo MA, Yang Y, Luo H, Giri S, Hart JJ, Faquin V, Guilherme LR,
Thannhauser TW, Li L (2016) Selenium promotes sulfur accumulation and plant growth in
wheat (Triticum aestivum). Physiol Plant 158:80–91
Brigelius-Flohé R, Maiorino M (2013) Glutathione peroxidases. Biochim Biophys Acta (BBA)
Gen Subj 1830(5):3289–3303
Brown T, Shrift A (1982) Selenium: toxicity and tolerance in higher plants. Biol Rev 57(1):59–84
Burnell JN (1981) Selenium metabolism in Neptunia amplexicaulis. Plant Physiol 67(2):316–324
Cabaraux JF, Dotreppe O, Hornick JL, Istasse L, Dufrasne I (2007) Les oligo-éléments dans
l'alimentation des ruminants: État des lieux, formes et efficacité des apports avec une atten-
tion particulière pour le sélénium. CRA-W-Fourrages Actualités, 12ème journée, 2007; 28–36.
Cartes P, Gianfreda L, Mora M (2005) Uptake of selenium and its antioxidant activity in ryegrass
when applied as selenate and selenite forms. Plant Soil 276(1–2):359–367
Das I, Pradhan M (2016) Potassium-solubilizing microorganisms and their role in enhancing
soil fertility and health. In: Meena VS, Maurya BR, Verma JP, Meena RS (eds) Potassium
solubilizing microorganisms for sustainable agriculture. Springer, India, pp 281–291.
doi:10.1007/978-81-322-2776-2_20
De Souza M, Huang C, Chee N, Terry N (1999) Rhizosphere bacteria enhance the accumulation of
selenium and mercury in wetland plants. Planta 209(2):259–263
Dixon J, Braun H, Crouch J (2009) Overview: transitioning wheat research to serve the future
needs of the developing world. Wheat facts and Futures 2009. Mexico: CIMMYT 23 (4.3):1
Dominguez-Nunez JA, Benito B, Berrocal-Lobo M, Albanesi A (2016) Mycorrhizal fungi: role
in the solubilization of potassium. In: Meena VS, Maurya BR, Verma JP, Meena RS (eds)
Potassium solubilizing microorganisms for sustainable agriculture. Springer, India, pp 77–98.
doi:10.1007/978-81-322-2776-2_6
Dotaniya ML, Meena VD, Basak BB, Meena RS (2016) Potassium uptake by crops as well
as microorganisms. In: Meena VS, Maurya BR, Verma JP, Meena RS (eds) Potassium
solubilizing microorganisms for sustainable agriculture. Springer, India, pp 267–280.
doi:10.1007/978-81-322-2776-2_19
Du Q, Yao H, Yao L, Zhang Z, Lei X, Xu S (2016) Selenium deficiency influences the expression of
selenoproteins and inflammatory cytokines in chicken aorta vessels. Biol Trace Elem Res:1–13
Ducsay L, Lozek O (2006) Effect of selenium foliar application on its content in winter wheat
grain. Plant Soil Environ 52(2):78
Durán P, Acuña J, Jorquera M, Azcón R, Borie F, Cornejo P, Mora M (2013) Enhanced selenium
content in wheat grain by co-inoculation of selenobacteria and arbuscular mycorrhizal fungi: a
preliminary study as a potential se biofortification strategy. J Cereal Sci 57(3):275–280
Durán P, Acuña JJ, Jorquera MA, Azcón R, Paredes C, Rengel Z, de la Luz MM (2014) Endophytic
bacteria from selenium-supplemented wheat plants could be useful for plant-growth promo-
tion, biofortification and Gaeumannomyces graminis biocontrol in wheat production. Biol
Fertil Soils 50(6):983–990
14 Bacterial-Mediated Selenium Biofortification of Triticum aestivum: Strategy… 311
L.) in the alluvial soil under indo-gangetic plain of India. J Plant Growth Reg. doi:10.1007/
s00344-016-9663-5
Leach A, Mumford J (2008) Pesticide environmental accounting: a method for assessing the exter-
nal costs of individual pesticide applications. Environ Pollut 151(1):139–147
Li HF, McGrath SP, Zhao FJ (2008) Selenium uptake, translocation and speciation in wheat sup-
plied with selenate or selenite. New Phytol 178(1):92–102
Lyons G, Ortiz-Monasterio I, Stangoulis J, Graham R (2005a) Selenium concentration in wheat
grain: is there sufficient genotypic variation to use in breeding? Plant Soil 269(1–2):369–380
Lyons GH, Judson GJ, Ortiz-Monasterio I, Genc Y, Stangoulis JC, Graham RD (2005b) Selenium
in Australia: selenium status and biofortification of wheat for better health. J Trace Elem Med
Biol 19(1):75–82
Masood S, Bano A (2016) Mechanism of potassium solubilization in the agricultural soils by
the help of soil microorganisms. In: Meena VS, Maurya BR, Verma JP, Meena RS (eds)
Potassium solubilizing microorganisms for sustainable agriculture. Springer, India, pp 137–
147. doi:10.1007/978-81-322-2776-2_10
Maurya BR, Meena VS, Meena OP (2014) Influence of Inceptisol and Alfisol’s potassium solubi-
lizing bacteria (KSB) isolates on release of K from waste mica. Vegetos 27:181–187
Mayland HF (1994) Selenium in plant and animal nutrition. In: Frankkenberger, WT BS (eds)
selenium in the environment. Marcel-Dekker, New York, pp 29–47
Meena OP, Maurya BR, Meena VS (2013a) Influence of K-solubilizing bacteria on release of
potassium from waste mica. Agric Sust Dev 1:53–56
Meena VS, Maurya BR, Bohra JS, Verma R, Meena MD (2013b) Effect of concentrate manure
and nutrient levels on enzymatic activities and microbial population under submerged rice in
alluvium soil of Varanasi. Crop Res. 45 (1,2 & 3): 6-12.
Meena VS, Maurya BR, Verma R, Meena RS, Jatav GK, Meena SK, Meena SK (2013c) Soil
microbial population and selected enzyme activities as influenced by concentrate manure and
inorganic fertilizer in alluvium soil of Varanasi. The Bioscan 8(3):931–935
Meena VS, Maurya BR, Bahadur I (2014a) Potassium solubilization by bacterial strain in waste
mica. Bang J Bot 43:235–237
Meena VS, Maurya BR, Verma JP (2014b) Does a rhizospheric microorganism enhance K+ avail-
ability in agricultural soils? Microbiol Res 169:337–347
Meena RS, Meena VS, Meena SK, Verma JP (2015a) The needs of healthy soils for a healthy
world. J Clean Prod 102:560–561
Meena RS, Meena VS, Meena SK, Verma JP (2015b) Towards the plant stress mitigate the agricul-
tural productivity: a book review. J Clean Prod 102:552–553
Meena VS, Maurya BR, Meena RS (2015c) Residual impact of wellgrow formulation and NPK on
growth and yield of wheat (Triticum aestivum L.) Bangladesh J. Bot 44(1):143–146
Meena VS, Maurya BR, Verma JP, Aeron A, Kumar A, Kim K, Bajpai VK (2015d) Potassium
solubilizing rhizobacteria (KSR): isolation, identification, and K-release dynamics from waste
mica. Ecol Eng 81:340–347
Meena VS, Meena SK, Verma JP, Meena RS, Ghosh BN (2015e) The needs of nutrient use efficiency
for sustainable agriculture. J Clean Prod 102:562–563. doi:10.1016/j.jclepro.2015.04.044
Meena VS, Verma JP, Meena SK (2015f) Towards the current scenario of nutrient use efficiency in
crop species. J Clean Prod 102:556–557. doi:10.1016/j.jclepro.2015.04.030
Meena RK, Singh RK, Singh NP, Meena SK, Meena VS (2016a) Isolation of low temperature sur-
viving plant growth-promoting rhizobacteria (PGPR) from pea (Pisum sativum L.) and docu-
mentation of their plant growth promoting traits. Biocatalysis Agric Biotechnol 4:806–811
Meena RS, Bohra JS, Singh SP, Meena VS, Verma JP, Verma SK, Sihag SK (2016b) Towards the
prime response of manure to enhance nutrient use efficiency and soil sustainability a current
need: a book review. J Clean Prod 112(1):1258–1260
Meena SK, Rakshit A, Meena VS (2016c) Effect of seed bio-priming and N doses under varied
soil type on nitrogen use efficiency (NUE) of wheat (Triticum aestivum L.) under greenhouse
conditions. Biocatalysis Agric Biotechnol 6:68–75
14 Bacterial-Mediated Selenium Biofortification of Triticum aestivum: Strategy… 313
Meena VS, Bahadur I, Maurya BR, Kumar A, Meena RK, Meena SK, Verma JP (2016d) Potassium-
solubilizing microorganism in evergreen agriculture: an overview. In: Meena VS, Maurya BR,
Verma JP, Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture.
Springer, India, pp 1–20. doi:10.1007/978-81-322-2776-2_1
Meena VS, Meena SK, Bisht JK, Pattanayak A (2016e) Conservation agricultural practices in
sustainable food production. J Clean Prod 137:690–691
Neal RH (1995) Selenium. In: Alloway BJ (ed) Heavy metals in soils. Blackie Academic &
Professional, London, pp 260–283
Nelson GC, Rosegrant MW, Palazzo A, Gray I, Ingersoll C, Robertson R, Tokgoz S, Zhu T, Sulser
TB, Ringler C (2010) Food security, farming, and climate change to 2050: scenarios, results,
policy options, vol 172. International Food Policy Research Institute, Washington, D C
Oancea F, Szabolcs L, Oancea AO, Lacatusu R, Abraham B, Stanciu-Burileanu MM, Meszaros A,
Lungu M (2014) Selenium biofortification biotechnologies of wheat grain in south-eastern part
of Romania for a better human health. Studia Universitatis “Vasile Goldis” Arad Seria Stiintele
Vietii (Life Sciences Series) 1:24–47
Parewa HP, Yadav J, Rakshit A, Meena VS, Karthikeyan N (2014) Plant growth promoting rhizo-
bacteria enhance growth and nutrient uptake of crops. Agric Sustain Dev 2(2):101–116
Pieczyńska J, Grajeta H (2015) The role of selenium in human conception and pregnancy. J Trace
Elem Med Biol 29:31–38
Prakash S, Verma JP (2016) Global perspective of potash for fertilizer production. In: Meena VS,
Maurya BR, Verma JP, Meena RS (eds) Potassium solubilizing microorganisms for sustainable
agriculture. Springer, India, pp 327–331. doi:10.1007/978-81-322-2776-2_23
Priyadharsini P, Muthukumar T (2016) Interactions between arbuscular mycorrhizal fungi and
potassium-solubilizing microorganisms on agricultural productivity. In: Meena VS, Maurya
BR, Verma JP, Meena RS (eds) Potassium solubilizing microorganisms for sustainable agricul-
ture. Springer, India, pp 111–125. doi:10.1007/978-81-322-2776-2_8
Raghavendra MP, Nayaka NC, Nuthan BR (2016) Role of rhizosphere microflora in potas-
sium solubilization. In: Meena VS, Maurya BR, Verma JP, Meena RS (eds) Potassium
solubilizing microorganisms for sustainable agriculture. Springer, India, pp 43–59.
doi:10.1007/978-81-322-2776-2_4
Rana A, Joshi M, Prasanna R, Shivay YS, Nain L (2012) Biofortification of wheat through inocula-
tion of plant growth promoting rhizobacteria and cyanobacteria. Eur J Soil Biol 50:118–126
Rawat J, Sanwal P, Saxena J (2016) Potassium and its role in sustainable agriculture. In: Meena
VS, Maurya BR, Verma JP, Meena RS (eds) Potassium solubilizing microorganisms for sus-
tainable agriculture. Springer, India, pp 235–253. doi:10.1007/978-81-322-2776-2_17
Ros G, Rotterdam A, Bussink D, Bindraban P (2016) Selenium fertilization strategies for bio-
fortification of food: an agro-ecosystem approach. Plant Soil 404(1–2):99–112
Rosen BP, Liu Z (2009) Transport pathways for arsenic and selenium: a minireview. Environ Int
35(3):512–515
Saha M, Maurya BR, Bahadur I, Kumar A, Meena VS (2016a) Can potassium-solubilising bac-
teria mitigate the potassium problems in India? In: Meena VS, Maurya BR, Verma JP, Meena
RS (eds) Potassium solubilizing microorganisms for sustainable agriculture. Springer, India,
pp 127–136. doi:10.1007/978-81-322-2776-2_9
Saha M, Maurya BR, Meena VS, Bahadur I, Kumar A (2016b) Identification and characterization
of potassium solubilizing bacteria (KSB) from indo-Gangetic plains of India. Biocatal Agric
Biotechnol 7:202–209
Sharma A, Shankhdhar D, Shankhdhar SC (2016) Potassium-solubilizing microorganisms:
mechanism and their role in potassium solubilization and uptake. In: Meena VS, Maurya BR,
Verma JP, Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture.
Springer, India, pp 203–219. doi:10.1007/978-81-322-2776-2_15
Shibagaki N, Rose A, McDermott JP, Fujiwara T, Hayashi H, Yoneyama T, Davies JP (2002)
Selenate-resistant mutants of Arabidopsis thaliana identify SULTR1; 2, a sulfate transporter
required for efficient transport of sulfate into roots. Plant J 29(4):475–486
314 Z.u. Abadin et al.
Shiferaw B, Smale M, Braun HJ, Duveiller E, Reynolds M, Muricho G (2013) Crops that feed
the world10. Past successes and future challenges to the role played by wheat in global food
security. Food Secure 5(3):291–317
Shrivastava M, Srivastava PC, D’Souza SF (2016) KSM soil diversity and mineral solubilization,
in relation to crop production and molecular mechanism. In: Meena VS, Maurya BR, Verma JP,
Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture. Springer,
India, pp 221–234. doi:10.1007/978-81-322-2776-2_16
Sindhu SS, Parmar P, Phour M, Sehrawat A (2016) Potassium-solubilizing microorganisms
(KSMs) and its effect on plant growth improvement. In: Meena VS, Maurya BR, Verma JP,
Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture. Springer,
India, pp 171–185. doi:10.1007/978-81-322-2776-2_13
Singh NP, Singh RK, Meena VS, Meena RK (2015) Can we use maize (Zea mays) rhizobacteria as
plant growth promoter? Vegetos 28(1):86–99. doi:10.5958/2229–4473.2015.00012.9
Singh M, Dotaniya ML, Mishra A, Dotaniya CK, Regar KL, Lata M (2016) Role of biofertilizers in
conservation agriculture. In: Bisht JK, Meena VS, Mishra PK, Pattanayak a (eds) Conservation
agriculture: an approach to combat climate change in Indian Himalaya. Springer, Singapore,
pp 113–134. doi:10.1007/978-981-10-2558-7_4
Sirko A, Hryniewicz M, Hulanicka D, Böck A (1990) Sulfate and thiosulfate transport in
Escherichia coli K-12: nucleotide sequence and expression of the cysTWAM gene cluster.
J Bacteriol 172(6):3351–3357
Sors T, Ellis D, Na G, Lahner B, Lee S, Leustek T, Pickering I, Salt D (2005a) Role of sulfur
assimilating enzymes in selenate reduction, tolerance and accumulation in Astragalus. Plant
J 42:785–792
Sors T, Ellis D, Salt D (2005b) Selenium uptake, translocation, assimilation and metabolic fate in
plants. Photosynth Res 86(3):373–389
Supriatin S, Weng L, Comans RN (2016) Selenium-rich dissolved organic matter determines selenium
uptake in wheat grown on Low-selenium arable land soils. Plant Soil 408 (1–2):73–94
Suttle NF (2010) Mineral Nutrition of Livestock, 4th edn. MPG Books Group, London, p 565
Teotia P, Kumar V, Kumar M, Shrivastava N, Varma A (2016) Rhizosphere microbes: potassium
solubilization and crop productivity-present and future aspects. In: Meena VS, Maurya BR,
Verma JP, Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture.
Springer, India, pp 315–325. doi:10.1007/978-81-322-2776-2_22
Turner RJ, Weiner JH, Taylor DE (1998) Selenium metabolism in Escherichia coli. Biometals
11(3):223–227
USGS (2011) 2010 minerals yearbook-selenium and tellurium. United States Geological Survey,
Reston
Van Loon L (2007) Plant responses to plant growth-promoting rhizobacteria. Eur J Plant Pathol
119(3):243–254
Velazquez E, Silva LR, Ramírez-Bahena MH, Peix A (2016) Diversity of potassium-solubilizing
microorganisms and their interactions with plants. In: Meena VS, Maurya BR, Verma JP,
Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture. Springer,
India, pp 99–110. doi:10.1007/978-81-322-2776-2_7
Verma R, Maurya BR, Meena VS (2014) Integrated effect of bio-organics with chemical fertilizer
on growth, yield and quality of cabbage (Brassica oleracea var capitata). Indian J Agric Sci
84(8):914–919
Verma JP, Jaiswa DK, Meena VS, Meena RS (2015a) Current need of organic farming for enhanc-
ing sustainable agriculture. J Clean Prod 102:545–547
Verma JP, Jaiswal DK, Meena VS, Kumar A, Meena RS (2015b) Issues and challenges about sus-
tainable agriculture production for management of natural resources to sustain soil fertility and
health. J Clean Prod 107:793–794
White P, Bowen H, Parmaguru P, Fritz M, Spracklen W, Spiby R, Meacham M, Mead A, Harriman
M, Trueman L (2004) Interactions between selenium and sulphur nutrition in Arabidopsis
thaliana. J Exp Bot 55(404):1927–1937
14 Bacterial-Mediated Selenium Biofortification of Triticum aestivum: Strategy… 315
World Health Organization (1996) Trace elements in human nutrition and health. World Health
Organization, Geneva
Yadav BK, Sidhu AS (2016) Dynamics of potassium and their bioavailability for plant nutrition.
In: Meena VS, Maurya BR, Verma JP, Meena RS (eds) Potassium solubilizing microorganisms
for sustainable agriculture. Springer, India, pp 187–201. doi:10.1007/978-81-322-2776-2_14
Yanai J, Mizuhara S, Yamada H (2015) Soluble selenium content of agricultural soils in Japan
and its determining factors with reference to soil type, land use and region. Soil Sci Plant Nutr
61(2):312–318
Yasin M, Faisal M (2013) Growth responses of Triticum aestivum after inoculating with
Pseudomonas and Stenotrophomonas. Afr J Microbiol Res 7(19):1952–1956
Yasin M, El-Mehdawi A, Anwar A, Pilon-Smits E, Faisal M (2015a) Microbial-enhanced selenium
and iron biofortification of wheat (Triticum aestivum L.)-applications in phytoremediation and
biofortification. Int J Phytoremediation 17(4):341–347
Yasin M, El-Mehdawi AF, Pilon-Smits EA, Faisal M (2015b) Selenium-fortified wheat: potential of
microbes for biofortification of selenium and other essential nutrients. Int J Phytoremediation
17(8):777–786
Yasin M, Munir I, Faisal M (2016) Can Bacillus spp. enhance K+ uptake in crop species. In:
Meena VS, Maurya BR, Verma JP, Meena RS (eds) Potassium solubilizing microorganisms
for sustainable agriculture. Springer, India, pp 163–170. doi:10.1007/978-81-322-2776-2_12
Zahedi H (2016) Growth-promoting effect of potassium-solubilizing microorganisms on
some crop species. In: Meena VS, Maurya BR, Verma JP, Meena RS (eds) Potassium
solubilizing microorganisms for sustainable agriculture. Springer, India, pp 31–42.
doi:10.1007/978-81-322-2776-2_3
Zhao J, Xing H, Liu C, Zhang Z, Xu S (2016) Effect of selenium deficiency on nitric oxide and
heat shock proteins in chicken erythrocytes. Biol Trace Elem Res 171(1):208–213
Role of Vesicular-Arbuscular
Mycorrhizae in Mobilization of Soil 15
Phosphorus
Abstract
The microorganisms play a vital role in sustaining the crop production through
improving the soil properties and plant nutrition. Among the microorganisms
VAM (vesicular-arbuscular mycorrhizae) is a beneficial fungus that plays an
important role in soil nutrient dynamics and improving soil physical, chemical
and biological properties. Though phosphorus is the second macronutrient
required in relatively large amounts by plants next to nitrogen, it is one of the
most difficult nutrients for plants to acquire because of its low solubility, low
mobility and fixation in soil. The mycorrhizal symbiotic association between
fungi and plants plays an important role in the uptake of phosphorus. Many
experiments have specified that VAM is able to alter mobilization of soil phos-
phorus of its host plants. The paper summarizes about mycorrhizal symbiosis of
VAM involving multistep colonization process, soil phosphorus dynamics in the
rhizosphere and mycorrhizal mechanism and pathways involved in phosphorus
availability and uptake.
Keywords
Vesicular-arbuscular mycorrhizae • Colonization process • Soil phosphorus
dynamics • Phosphorus availability and uptake
15.1 Introduction
The microorganisms play a vital role in sustaining the crop production through
improving the soil properties and plant nutrition. Though the agricultural produc-
tion has been continued to increase to feed the blooming population in one side, the
soil fertility has attained the statue and started to decline because of faulty agricul-
tural practices. Because of that fertile lands are going out of cultivation and increased
degradation results increased area under current fallows. In this situation, we must
go for a combined use of both organic and inorganic fertilizers in judicious combi-
nation based on soil nutrient status to improve and sustain the soil fertility and
productivity which is called as integrated nutrient management (INM). One of the
major components in INM practices is the use of bio-fertilizers which are prepared
from many beneficial microbial inoculants (Meena et al. 2013a, 2016a; Bahadur
et al. 2014; Maurya et al. 2014; Jat et al. 2015; Kumar et al. 2015, 2016b; Ahmad
et al. 2016).
The main beneficial activities of microorganisms are increased availability of
plant nutrients, improvement of nutrient uptake, production of plant growth regula-
tors, plant protection against root pathogens etc. According to their relationships
with the plant, the microorganisms can be divided into saprophytes, parasitic
symbionts (pathogen) and mutualistic symbionts (symbionts). The mutualistic sym-
bionts are commonly used in bio-fertilizers because it carries out functions for the
plants that they are unable to perform for themselves. For example, the host plant
receives mineral nutrients by the organisms, and the microorganism obtains photo-
synthetically derived carbon compounds from the host plant (Finlay 2008). Among
these the microorganism VAM is a beneficial fungus that plays an important role
in soil nutrient dynamics and improving soil physical, chemical and biological
properties (Parewa et al. 2014; Prakash and Verma 2016; Meena et al. 2016b;
Jaiswal et al. 2016; Jha and Subramanian 2016; Kumar et al. 2016a).
Frank was the first person to coin the term “Mykorrhizen” in the year 1885 for fun-
gal association in the trees of pines. Mykorrhizen is a Greek term meaning “fungal
root”. Paleobotany dates these associations to the Devonian era (~400 million years
back). These mycorrhizal associations played predominant role in the transforma-
tion of primitive aquatic plants to highly evolved terrestrial plants during the course
of evolution. Today, mycorrhiza is the most widespread symbiotic association exist-
ing in the ecosystems throughout the world (Priyadharsini and Muthukumar 2016;
Kumar et al. 2017; Meena et al. 2015a, b, f; Raghavendra et al. 2016; Zahedi 2016;
Dominguez-Nunez et al. 2016; Dotaniya et al. 2016).
Members of several fungal taxa are involved in mycorrhizal associations and
possess a few common characteristic features such as ubiquitous distribution,
strong biotrophic dependence on host plants and rarely free-living saprophytes.
To date, seven types of mycorrhizae are recognized, viz. ectomycorrhiza,
15 Role of Vesicular-Arbuscular Mycorrhizae in Mobilization of Soil Phosphorus 319
cortex
Hyphal branching
Ar
Hyphal fans
root
External hyphae
spore formation
Fig. 15.1 Mycorrhizal multistep colonization (Adapted from Gadkar et al. 2011)
2013b, 2014a, 2015e, 2016d, e; Sharma et al. 2016; Verma et al. 2015a; Shrivastava
et al. 2016; Bahadur et al. 2016a; Masood and Bano 2016; Teotia et al. 2016). The
factors which influence the mycorrhizal association are host plant, mycorrhizal
fungi, soil and environmental conditions (Barea 1991). If any one of these factors is
inhibitory to mycorrhizal fungi, the symbiosis will be inhibited even if other factors
are optimum (Liu et al. 2000).
The adequate amount of soil phosphorus is required to enhance shoot and root
growth and promote early maturity which in turn help to increase water use
efficiency (WUE) and yield potential because it is the element, which helps
to store and transfer energy produced by photosynthesis (Tairo and Ndakidemi
2013). Though phosphorus is the second macronutrient required in relatively large
amounts by plants next to nitrogen, it is one of the most difficult nutrients for plants
to acquire because of its low solubility, low mobility and fixation in soil (Balemi and
Negisho 2012).
The availability of soil phosphorus is affected by the presence of Fe and Al
oxides which fix phosphorus as Fe and Al phosphates and clay minerals in highly
weathered alfisol, ultisol and oxisols and as Ca phosphates under calcareous soils
(Amanullah et al. 2010). Despite its availability, the phosphorus uptake is influ-
enced by the rooting type, soil properties and soil moisture availability. Since the
P availability depends on many factors, most of the plant phosphorus requirement
15 Role of Vesicular-Arbuscular Mycorrhizae in Mobilization of Soil Phosphorus 321
has been met by diffusion process rather than by mass flow and root interception. In
the diffusion process, once the phosphate is adsorbed strongly by the soil, its effec-
tive diffusion coefficient value becomes low, which creates a steep concentration
gradient in the soil (Kanno et al. 2016). Hence the soil phosphate near the root zone
only could reach the root surface to meet the plant requirements and ultimately plant
express phosphorus deficiency when the demand is more than the availability exist
in soil.
The mycorrhizal symbiotic association between fungi and plants plays an important
role in the uptake of phosphorus. Phosphorus has very limited mobility in soils and
despite of its low availability it has been utilized quickly around roots and form
depletion zones. Thus to obtain more phosphorus, plants must bypass these deple-
tion zones by further root activity. The increased P uptake of mycorrhizal plants is
mainly due to absorption and translocation of P from distant areas, which are other-
wise inaccessible to plant roots. Hyphae absorb P in the form of orthophosphate and
transported actively as polyphosphates (Tinker and Nye 2000; Hinsinger 2001;
Plassard and Dell 2010).
The increase in absorption of P by mycorrhizal plants has been attributed to
increase in surface area for absorption (Smith and Read 2008; Smith and Smith
2011a). The fineness of hyphae has twofold advantages over root hairs by increase
in the surface area of hyphae for greater absorption of nutrient and increase the area
of exploration by enabling the entry of hyphae into soil pores which cannot be
entered by root hairs. Rapid absorption of soluble form of P by the extra radical
hyphae leads to a shift in the equilibrium towards the release of bound P from the
soil reserves. It has often been observed that mycorrhizal roots on a unit weight
basis absorbed much higher amounts of P than non-mycorrhizal plants. This sug-
gests that mycorrhizal fungus hyphae have higher affinity for phosphate ions at
lower threshold concentration for absorption than do plant roots (Bolan 1991).
There are two possible ways in which soil phosphorus has been taken up by the
plants: (i) direct pathway and (ii) AM pathway (Smith et al. 2011). Generally plants
uptake phosphorus through direct pathway as negatively charged H2PO4− ions which
increase negative electric potential inside the cell membrane because of high cell
concentration compared to soil solution. Hence an additional metabolic energy
required for the Pi uptake, and it needs high-affinity transporter proteins. So the
pathway is more effective in the root apex which has high-affinity transporter
proteins (PiTs) in the epidermis. But after a certain period of time, two things will
happen. The first is when loss of root hairs started, the transporter protein (PiT)
322 M. Lalitha et al.
3. P is transferred from
fungus to plant at intra-
cellular interfaces via AM-
inducible Pi transporters
Fig. 15.2 The phosphorus uptake pathways of mycorrhizal root (Adapted from Smith et al. 2011)
activity will reduce, and certainly direct uptake will decline. Second, the uptake of
phosphorus as orthophosphate (Pi) by root epidermal cells leads to lower the Pi
concentrations in the root rhizosphere making zone of depletion (Smith and Smith
2011b; Balzergue et al. 2011).
Accordingly plants developed a range of strategies to enhance Pi uptake capacity
(Marschner 1995) and among one is AM pathway (Fig. 15.2) in which soil phospho-
rus is foraged from large volumes of soil and transported to cortical cells evaded
direct uptake. In the mycorrhizal pathway AM fungal hyphae uptake the orthophos-
phate (Pi) by fungal Pi transporters from the root and directly translocated into the
intracellular fungal structures of arbuscules and hyphal coils which is present in root
cortical cells (Smith et al. 2011). The fungal Pi transporters are specific to AM path-
way, and it is different from the direct uptake expressed exclusively in the colonized
cortical cell. The plant Pi transporters induced in colonized cells transfer Pi from the
interfacial apoplast to the plant cortical cells and meet the plant requirement
(Velazquez et al. 2016; Meena et al. 2013c, 2014b, 2015c, d; Sindhu et al. 2016;
Singh et al. 2015, 2016).
The association between concentrations of extractable soil phosphate and the extent
of mycorrhizal colonization in plants are ambiguous. Sites with large amounts of
extractable phosphate may have high levels of infection and large numbers of
spores, whereas sites with small amounts of extractable phosphate may have low
levels of colonization or low numbers of spores (Gianinazzi-Pearson et al. 1980;
Almagrabi and Abdelmoneim 2012). In contradictory Rathore and Singh (1995)
and Ezawa et al. (2005) reported that plant inoculation with VAM in the absence of
added P significantly increased the available soil P because of the release of organic
15 Role of Vesicular-Arbuscular Mycorrhizae in Mobilization of Soil Phosphorus 323
Table 15.1 Available soil phosphorus (mg kg−1) under maize crop at 45 DAS as influenced by
VAM inoculation
Phosphorus doses (kg ha−1)
Mycorrhizal strain 0 15 30 45 60 Mean
Uninoculated 5.1 6.5 8.3 8.8 10.5 7.9
G. margarita 5.8 6.5 8.8 10.6 11.4 8.6
G. fasciculatum 6.5 7.5 8.8 11.3 12.6 9.4
VAM mixed endophyte 6.0 7.5 8.4 10.9 11.5 8.9
CD(P = 0.01) M 0.1 P 0.1 M × P 0.2
Rathore and Singh (1995)
exudates in the rhizosphere and concluded that mycorrhizal inoculam may substi-
tute phosphatic fertilizer equivalent of 30 kg P ha−1 (Table 15.1).
However, in some studies a negative association has been found between the
amounts of extractable phosphate in soils and the abundance of VAM fungi as
assessed by infection (Liu et al. 2000). Adding phosphate fertilizers has been shown
either to have no effect (Kahiluoto et al. 2001) or to decrease (Ryan and Graham
2002) the level of mycorrhizal infection in a range of agricultural crops. This
relationship is probably correlated with effects of phosphate on lengths of root
colonized. The low percentage of root length colonized at high phosphorus avail-
ability does not necessarily imply plant suppression or control of fungal activity;
there may be no effects of high phosphorus on the fungus per se (Marschner 1995;
Mathimaran et al. 2005; Beauregard et al. 2010). But very high phosphorus applica-
tion can definitely alter root colonization particularly reducing arbuscule develop-
ment and decrease mycorrhizal fungal biomass per plant (Smith and Read 2008).
In addition reductions in appressorium formation were also observed in Pisum
sativum roots at high phosphorus which was arbitrated by internal plant-derived
signals (Balzergue et al. 2011). When phosphorus is abundant, a direct possibly less
costly uptake pathway is preferred and the rate of root colonization also low (Nagy
et al. 2008). However, a group of secondary metabolites called strigolactones
which mediate signalling necessary for root colonization has strong negative effect
with high phosphorus supply in various species (Lopez-Raez et al. 2008) which is
also one of the reasons for the less colonization. Furthermore, the effect of soil
phosphorus on mycorrhizal fungal diversity is also confounded by factors such as
host species (Gosling et al. 2013).
The sources of different phosphorus fertilizer do not have any significant effect on
mycorrhizal infection when it was applied in low rate. The effect of phosphorus
source is evident where the rate of fertilizer application is high. The high root length
infection is always associated with rock phosphate compared to superphosphate.
Phosphate in the soil solution derived from the Ghafsa phosphate rock source is
expected to be different from the superphosphate source, and so the difference in
324 M. Lalitha et al.
infectivity associated with the higher rate of superphosphate application could also
have been due to the differences in the rates of dissolution of superphosphate and
phosphate rock to provide phosphorus in the soil solution over a given period
(Asmah 1995).
It is believed that soil phosphorus has antagonistic interactions with Zn, when any
one of the nutrients available in soil is in excess (Gianquinto et al. 2000), and such
a relationship is a result of the dilution effect of plant growth due to improved P
nutrition. Further a study by Lu et al. (1998) demonstrated an increase in P avail-
ability caused no significant change in Zn concentrations of Brassica napus, and
interactions of P and Zn in Oryza sativa and Triticum aestivum were additive and
beneficial to the concentrations of P and Zn (Islam et al. 2005). The interaction
remains uncertain, and the presence or absence of mycorrhizal association on the
relationship was hypothesized, despite the fact that much literature has indicted that
mycorrhizal fungal colonization promotes P or Zn nutrition of host plants indepen-
dently. The high avaialble P (P addition up to 40 mg kg−1) in soil has increased plant
available zinc, whereas at higher P levels, the zinc translocation was inhibited (Rupa
et al. 2003) (Table 15.2).
There was a synergistic interaction between P and Zn, which had resulted in
extensive root growth in colonized plants. There is a strong positive relationship
between Zn and P contents in the roots (r = 0.95; P < 0.001) and also in shoots
(r = 0.96; P < 0.001) (Subramanian et al. 2008).
Table 15.2 Effects of different rates and sources of phosphorus application on root infection,
nutrient concentration and uptake of soil phosphorus in maize crop (Asmah 1995)
Root infection Nutrient concentration Nutrient uptake
Soil Treatment (%) (μg g−1) (mg/pot)
Paleustal Control 47.50 1.26 1.65
TSP1 78.30 1.27 2.58
TSP2 52.30 2.18 4.38
PR1 68.90 1.61 2.71
PR2 75.00 1.61 3.73
Haplustoll Control 58.00 0.99 1.12
TSP1 63.80 1.40 2.81
TSP2 50.00 1.37 3.21
PR1 77.60 1.12 2.18
PR2 80.25 1.16 2.33
LSD (P < 0.01) 16.48 0.39 0.68
TSP1 triple superphosphate at 44 kg P ha−l, TSP2 triple superphosphate at 22 kg P ha−1, PR1 rock
phosphate at 44 kg P ha−1, PR2 rock phosphate at 22 kg P ha−1
15 Role of Vesicular-Arbuscular Mycorrhizae in Mobilization of Soil Phosphorus 325
15.4.3 Phosphatase
Soil phosphatase plays an important role in the P nutrition of plants because it medi-
ates the release of inorganic phosphorus from organically bound phosphorus.
Mycorrhizal colonization has been shown to influence the phosphatase activity, par-
ticularly the increased alkaline phosphatase activity in the presence of mycorrhizal
hyphae (Tarafdar and Marschner 1994). Acid phosphatases have also been reported
in mycorrhizal fungi, and although their function is unclear, they may be associated
with the growth and development of the fungus within the host tissue as well as with
phosphorus acquisition in the rhizosphere. Subramanian et al. (2009) reported that
acid phosphatase activity of VAM-colonized soils was higher irrespective of
the stages of observation, but the increase was well exhibited at early stages than in
the later stages.
Plant water use efficiency has been accounted to decrease, increase or remain
unchanged with mycorrhizal colonization depending on plant or fungal species com-
bination considered (Auge 2001). The positive effects of mycorrhizal colonization
on plant water relations could be explained simply in terms of the improved phos-
phorus nutrition and larger biomass size of mycorrhizal plants. Increased phosphorus
uptake promoted root growth, which in turn enhanced the hydraulic conductivity and
transpiration rate in mycorrhizal colonization plants (Bethlenfalvay et al. 1988).
Hence the contribution of mycorrhizal colonization towards plant drought tolerance
may be due to the impact of accumulative physical, nutritional, physiological and
cellular effects imposed by vesicular-arbuscular mycorrhizae (Davies et al. 1992).
15.6 Conclusions
The fertilizer phosphate is becoming one of the most expensive commodities, and its
availability is uncertain; the crop production will be critical, particularly in rainfed
regions, where the fertilizer application is mainly based on rainfall distribution. In
this situation mycorrhizal plant colonization is very useful in conserving energy by
reducing fertilizer requirement of crops and in meeting the production targets in
nutritionally deficient soils. The omnipresent nature, increased phosphorous uptake,
drought tolerance and increased plant growth benefit by mycorrhizal colonization,
which is important to increase the absorption of other relatively immobile elements
in soil and a step towards the sustainability of the healthy soil and plant ecosystem.
Acknowledgements We thank the editors and anonymous reviewers for their constructive com-
ments, which helped us to improve the manuscript.
326 M. Lalitha et al.
References
Ahmad M, Nadeem SM, Naveed M, Zahir ZA (2016) Potassium-solubilizing bacteria and their
application in agriculture. In: Meena VS, Maurya BR, Verma JP, Meena RS (eds) Potassium
solubilizing microorganisms for sustainable agriculture. Springer, New Delhi, pp 293–313.
doi:10.1007/978-81-322-2776-2_21
Allen MF (1991) The ecology of mycorrhizae. Cambridge University Press, Cambridge, p 184
Almagrabi OA, Abdelmoneim TS (2012) Using of arbuscular mycorrhizal fungi to reduce
the deficiency effect of phosphorous fertilization on maize plants (Zea mays L.) Life Sci
J 9(4):1648–1654
Amanullah, Zakirullah M, Khalil SK (2010) Timing and rate of phosphorus application influence
maize phenology, yield and profitability in Northwest Pakistan. Int J Plant Prod 4:281–292
Asmah AE (1995) Effect of phosphorus source and rate of application on VAM fungal infection
and growth of maize (Zea mays L.). Mycorrhiza 5(3):223–228
Auge RM (2001) Water relations, drought and vesicular-arbuscular mycorrhizal symbiosis.
Mycorrhiza 11:3–42
Bahadur I, Meena VS, Kumar S (2014) Importance and application of potassic biofertilizer in
Indian agriculture. Int Res J Biol Sci 3:80–85
Bahadur I, Maurya BR, Kumar A, Meena VS, Raghuwanshi R (2016a) Towards the soil sustain-
ability and potassium-solubilizing microorganisms. In: Meena VS, Maurya BR, Verma JP,
Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture. Springer,
New Delhi, pp 225–266. doi:10.1007/978-81-322-2776-2_18
Bahadur I, Maurya BR, Meena VS, Saha M, Kumar A, Aeron A (2016b) Mineral release dynamics
of tricalcium phosphate and waste muscovite by mineral-solubilizing rhizobacteria isolated
from indo-gangetic plain of India. Geomicrobiol J. doi:10.1080/01490451.2016.1219431
Balemi T, Negisho K (2012) Management of soil phosphorus and plant adaptation mechanisms to
phosphorus stress for sustainable crop production: a review. J Soil Sci Plant Nutr 12(3):547–562
Balzergue C, Puech Pages V, Becard G, Rochange SF (2011) The regulation of arbuscular mycor-
rhizal symbiosis by phosphate in pea involves early and systemic signalling events. J Exp Bot
62:1049–1060
Barea JM (1991) Vesicular-arbuscular mycorrhizae as modifiers of soil fertility. In: Stewart BA
(ed) Advances in soil science. Springer, New York, pp l–40
Beauregard MS, Hamel C, Atul-Nayyar St-Arnaud M (2010) Long-term phosphorus fertilization
impacts soil fungal and bacterial diversity but not AM fungal community in alfalfa. Microb
Ecol 59:379–389
Bethlenfalvay GJ, Thomas RS, Dakessian S, Brown MS, Ames RN, Whitehead EE (1988)
Mycorrhizae in stressed environments: effects on plant growth, endophyte development, soil
stability and soil water. In: Hutchinson CF, Timmermann BN (eds) Arid lands: today and
tomorrow. Westview, Boulder, pp 1015–1029
Bolan NS (1991) A critical review on the role of mycorrhizal fungi in the uptake of phosphorus by
plants. Plant Soil 134:189–207
Brundrett MC (2009) Mycorrhizal associations and other means of nutrition of vascular plants:
understanding the global diversity of host plants by resolving conflicting information and
developing reliable means of diagnosis. Plant Soil 320:37–77
Das I, Pradhan M (2016) Potassium-solubilizing microorganisms and their role in enhancing
soil fertility and health. In: Meena VS, Maurya BR, Verma JP, Meena RS (eds) Potassium
solubilizing microorganisms for sustainable agriculture. Springer, New Delhi, pp 281–291.
doi:10.1007/978-81-322-2776-2_20
Davies FT, Potter JR, Linderman RG (1992) Mycorrhiza and repeated drought exposure affect
drought resistance and extraradical hyphae development of pepper plants independent of plant
size and nutrient content. J Plant Physiol 139:289–294
Dominguez-Nunez JA, Benito B, Berrocal-Lobo M, Albanesi A (2016) Mycorrhizal fungi: role
in the solubilization of potassium. In: Meena VS, Maurya BR, Verma JP, Meena RS (eds)
15 Role of Vesicular-Arbuscular Mycorrhizae in Mobilization of Soil Phosphorus 327
Kumar A, Maurya BR, Raghuwanshi R, Meena VS, Islam MT (2017) Co-inoculation with
Enterobacter and Rhizobacteria on yield and nutrient uptake by wheat (Triticum aestivum L.)
in the alluvial soil under indo-gangetic plain of India. J Plant Growth Regul. doi:10.1007/
s00344-016-9663-5
Liu AC, Hamel RI, Hamilton SDL (2000) Mycorrhizae formation and nutrient uptake of new corn
(Zea mays L.) hybrids with extreme canopy and leaf architecture as influenced by soil N and P
levels. Plant Soil 221:157–166
Lopez-Raez JA, Charnikhova T, Gomez-Roldan V, Matusova R, Kohlen W, De Vos R, Verstappen
F, Puech-Pages V, Becard G, Mulder P, Bouwmeester H (2008) Tomato strigolactones are
derived from carotenoids and their biosynthesis is promoted by phosphate starvation. New
Phytol 178:863–874
Lu ZG, Grewal HS, Graham RD (1998) Dry matter production and uptake of zinc and phosphorus
in two oilseed rape genotypes under differential rates of zinc and phosphorus supply. J Plant
Nutr 21:25–38
Marschner H (1995) Mineral nutrition of higher plants, 2nd edn. Academic, London
Masood S, Bano A (2016) Mechanism of potassium solubilization in the agricultural soils by the
help of soil microorganisms. In: Meena VS, Maurya BR, Verma JP, Meena RS (eds) Potassium
solubilizing microorganisms for sustainable agriculture. Springer, New Delhi, pp 137–147.
doi:10.1007/978-81-322-2776-2_10
Mathimaran N, Ruh R, Vullioud P, Frossard E, Jansa J (2005) Glomus intraradices dominates
arbuscular mycorrhizal communities in a heavy textured agricultural soil. Mycorrhiza 16:61–66
Maurya BR, Meena VS, Meena OP (2014) Influence of Inceptisol and Alfisol’s potassium solubi-
lizing bacteria (KSB) isolates on release of K from waste mica. Vegetos 27:181–187
Meena OP, Maurya BR, Meena VS (2013a) Influence of K-solubilizing bacteria on release of
potassium from waste mica. Agric Sustain Dev 1:53–56
Meena VS, Maurya BR, Bohra JS, Verma R, Meena MD (2013b) Effect of concentrate manure
and nutrient levels on enzymatic activities and microbial population under submerged rice in
alluvium soil of Varanasi. Crop Res 45(1, 2 & 3):6–12
Meena VS, Maurya BR, Verma R, Meena RS, Jatav GK, Meena SK, Meena SK (2013c) Soil
microbial population and selected enzyme activities as influenced by concentrate manure and
inorganic fertilizer in alluvium soil of Varanasi. Bioscan 8(3):931–935
Meena VS, Maurya BR, Bahadur I (2014a) Potassium solubilization by bacterial strain in waste
mica. Bangladesh J Bot 43:235–237
Meena VS, Maurya BR, Verma JP (2014b) Does a rhizospheric microorganism enhance K+ avail-
ability in agricultural soils? Microbiol Res 169:337–347
Meena RS, Meena VS, Meena SK, Verma JP (2015a) The needs of healthy soils for a healthy
world. J Clean Prod 102:560–561
Meena RS, Meena VS, Meena SK, Verma JP (2015b) Towards the plant stress mitigate the agricul-
tural productivity: a book review. J Clean Prod 102:552–553
Meena VS, Maurya BR, Meena RS (2015c) Residual impact of wellgrow formulation and NPK on
growth and yield of wheat (Triticum aestivum L.) Bangladesh J Bot 44(1):143–146
Meena VS, Maurya BR, Verma JP, Aeron A, Kumar A, Kim K, Bajpai VK (2015d) Potassium
solubilizing rhizobacteria (KSR): isolation, identification, and K-release dynamics from waste
mica. Ecol Eng 81:340–347
Meena VS, Meena SK, Verma JP, Meena RS, Ghosh BN (2015e) The needs of nutrient use efficiency
for sustainable agriculture. J Clean Prod 102:562–563. doi:10.1016/j.jclepro.2015.04.044
Meena VS, Verma JP, Meena SK (2015f) Towards the current scenario of nutrient use efficiency in
crop species. J Clean Prod 102:556–557. doi:10.1016/j.jclepro.2015.04.030
Meena RK, Singh RK, Singh NP, Meena SK, Meena VS (2016a) Isolation of low temperature sur-
viving plant growth-promoting rhizobacteria (PGPR) from pea (Pisum sativum L.) and docu-
mentation of their plant growth promoting traits. Biocatalysis Agric Biotechnol 4:806–811
15 Role of Vesicular-Arbuscular Mycorrhizae in Mobilization of Soil Phosphorus 329
Meena RS, Bohra JS, Singh SP, Meena VS, Verma JP, Verma SK, Sihag SK (2016b) Towards the
prime response of manure to enhance nutrient use efficiency and soil sustainability a current
need: a book review. J Clean Prod 112(1):1258–1260
Meena SK, Rakshit A, Meena VS (2016c) Effect of seed bio-priming and N doses under varied
soil type on nitrogen use efficiency (NUE) of wheat (Triticum aestivum L.) under greenhouse
conditions. Biocatal Agric Biotechnol 6:68–75
Meena VS, Bahadur I, Maurya BR, Kumar A, Meena RK, Meena SK, Verma JP (2016d) Potassium-
solubilizing microorganism in evergreen agriculture: an overview. In: Meena VS, Maurya BR,
Verma JP, Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture.
Springer, New Delhi, pp 1–20. doi:10.1007/978-81-322-2776-2_1
Meena VS, Meena SK, Bisht JK, Pattanayak A (2016e) Conservation agricultural practices in
sustainable food production. J Clean Prod 137:690–691
Nagy R, Drissner D, Amrhein N, Jakobsen I, Bucher M (2008) Mycorrhizal phosphate uptake
pathway in tomato is phosphorus-repressible and transcriptionally regulated. New Phytol
181:950–959
Parewa HP, Yadav J, Rakshit A, Meena VS, Karthikeyan N (2014) Plant growth promoting rhizo-
bacteria enhance growth and nutrient uptake of crops. Agric Sustain Dev 2(2):101–116
Plassard C, Dell B (2010) Phosphorus nutrition of mycorrhizal trees. Tree Physiol 30(9):1129–
1139. doi:10.1093/treephys/tpq063
Prakash S, Verma JP (2016) Global perspective of potash for fertilizer production. In: Meena VS,
Maurya BR, Verma JP, Meena RS (eds) Potassium solubilizing microorganisms for sustainable
agriculture. Springer, New Delhi, pp 327–331. doi:10.1007/978-81-322-2776-2_23
Priyadharsini P, Muthukumar T (2016) Interactions between arbuscular mycorrhizal fungi and
potassium-solubilizing microorganisms on agricultural productivity. In: Meena VS, Maurya
BR, Verma JP, Meena RS (eds) Potassium solubilizing microorganisms for sustainable agricul-
ture. Springer, New Delhi, pp 111–125. doi:10.1007/978-81-322-2776-2_8
Raghavendra MP, Nayaka NC, Nuthan BR (2016) Role of rhizosphere microflora in potas-
sium solubilization. In: Meena VS, Maurya BR, Verma JP, Meena RS (eds) Potassium
solubilizing microorganisms for sustainable agriculture. Springer, New Delhi, pp 43–59.
doi:10.1007/978-81-322-2776-2_4
Rathore VP, Singh HP (1995) Influence of vesicular-arbuscular mycorrhizal fungi and phosphate
on maize. J Indian Soc Soil Sci 43(2):207–210
Rawat J, Sanwal P, Saxena J (2016) Potassium and its role in sustainable agriculture. In: Meena
VS, Maurya BR, Verma JP, Meena RS (eds) Potassium solubilizing microorganisms for sus-
tainable agriculture. Springer, New Delhi, pp 235–253. doi:10.1007/978-81-322-2776-2_17
Rupa TR, Rao CS, Subba Rao A, Singh M (2003) Effects of farmyard manure and phospho-
rus on zinc transformations and phyto-availability in two alfisols of India. Bioresour Technol
87(3):279–288
Ryan MH, Graham JH (2002) Is there a role for arbuscular mycorrhizal fungi in production agri-
culture? Plant Soil 244:263–271
Saha M, Maurya BR, Bahadur I, Kumar A, Meena VS (2016a) Can potassium-solubilising bacteria
mitigate the potassium problems in India? In: Meena VS, Maurya BR, Verma JP, Meena RS
(eds) Potassium solubilizing microorganisms for sustainable agriculture. Springer, New Delhi,
pp 127–136. doi:10.1007/978-81-322-2776-2_9
Saha M, Maurya BR, Meena VS, Bahadur I, Kumar A (2016b) Identification and characterization
of potassium solubilizing bacteria (KSB) from Indo-Gangetic Plains of India. Biocatal Agric
Biotechnol 7:202–209
Sharma A, Shankhdhar D, Shankhdhar SC (2016) Potassium-solubilizing microorganisms:
mechanism and their role in potassium solubilization and uptake. In: Meena VS, Maurya BR,
Verma JP, Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture.
Springer, New Delhi, pp 203–219. doi:10.1007/978-81-322-2776-2_15
330 M. Lalitha et al.
Shrivastava M, Srivastava PC, D’Souza SF (2016) KSM soil diversity and mineral solubilization,
in relation to crop production and molecular mechanism. In: Meena VS, Maurya BR, Verma JP,
Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture. Springer,
New Delhi, pp 221–234. doi:10.1007/978-81-322-2776-2_16
Sindhu SS, Parmar P, Phour M, Sehrawat A (2016) Potassium-solubilizing microorganisms
(KSMs) and its effect on plant growth improvement. In: Meena VS, Maurya BR, Verma JP,
Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture. Springer,
New Delhi, pp 171–185. doi:10.1007/978-81-322-2776-2_13
Singh NP, Singh RK, Meena VS, Meena RK (2015) Can we use maize (Zea mays) rhizobacteria as
plant growth promoter? Vegetos 28(1):86–99. doi:10.5958/2229-4473.2015.00012.9
Singh M, Dotaniya ML, Mishra A, Dotaniya CK, Regar KL, Lata M (2016) Role of biofertilizers in
conservation agriculture. In: Bisht JK, Meena VS, Mishra PK, Pattanayak A (eds) Conservation
agriculture: an approach to combat climate change in Indian Himalaya. Springer, Singapore,
pp 113–134. doi:10.1007/978-981-10-2558-7_4
Smith SE, Read DJ (1997) Vesicular-arbuscular mycorrhizas. In: Mycorrhizal symbiosis. (2nd ed)
Academic, New York, pp 9–126
Smith SE, Read DJ (2008) Mycorrhizal symbiosis, 3rd edn. Academic, New York
Smith SE, Smith FA (1990) Structure and function of the interfaces in biotrophic symbioses as
they relate to nutrient transport. New Phytol 114:l–38
Smith SE, Smith FA (2011a) Fresh perspectives on the roles of arbuscular mycorrhizal fungi
in plant nutrition and growth In Press at Mycologia, published on September 20, 2011 as
doi:10.3852/11-229
Smith SE, Smith FA (2011b) Roles of arbuscular mycorrhizas in plant nutrition and growth: new
paradigms from cellular to ecosystems scales. Annu Rev Plant Biol 63:227–250
Smith SE, Jakobsen I, Gronlund M, Andrew Smith F (2011) Roles of arbuscular mycorrhizas in
plant phosphorus nutrition: interactions between pathways of phosphorus uptake in arbuscular
mycorrhizal roots have important implications for understanding and manipulating plant phos-
phorus acquisition. Plant Physiol 156:1050–1057
Subramanian KS, Bharathi C, Jegan A (2008) Response of maize to mycorrhizal colonization at
varying levels of zinc and phosphorus. Biol Fertil Soils 45:133–144
Subramanian KS, Tenshia V, Jayalakshmi K, Ramachandran V (2009) Biochemical changes and
zinc fractions in arbuscular mycorrhizal fungus (Glomus intraradices) inoculated and uninocu-
lated soils under differential zinc fertilization. Appl Soil Ecol 43:32–39
Tairo EV, Ndakidemi PA (2013) Possible benefits of rhizobial inoculation and phosphorus sup-
plementation on nutrition, growth and economic sustainability in grain legumes. Am J Res
Commun 1(12):532–556
Tarafdar JC, Marschner H (1994) Phosphatase activity in the rhizosphere and hyphosphere of
VA-mycorrhizal wheat supplied with inorganic and organic phosphorus. Soil Biol Biochem
26:387–395
Teotia P, Kumar V, Kumar M, Shrivastava N, Varma A (2016) Rhizosphere microbes: potassium
solubilization and crop productivity-present and future aspects. In: Meena VS, Maurya BR,
Verma JP, Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture.
Springer, New Delhi, pp 315–325. doi:10.1007/978-81-322-2776-2_22
Tinker PB, Nye PH (2000) Solute movement in the rhizosphere. Oxford University Press, New York
Velazquez E, Silva LR, Ramírez-Bahena MH, Peix A (2016) Diversity of potassium-solubilizing
microorganisms and their interactions with plants. In: Meena VS, Maurya BR, Verma JP,
Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture. Springer,
New Delhi, pp 99–110. doi:10.1007/978-81-322-2776-2_7
Verma R, Maurya BR, Meena VS (2014) Integrated effect of bio-organics with chemical fertilizer
on growth, yield and quality of cabbage (Brassica oleracea var capitata). Indian J Agric Sci
84(8):914–919
Verma JP, Jaiswa DK, Meena VS, Meena RS (2015a) Current need of organic farming for enhancing
sustainable agriculture. J Clean Prod 102:545–547
15 Role of Vesicular-Arbuscular Mycorrhizae in Mobilization of Soil Phosphorus 331
Verma JP, Jaiswal DK, Meena VS, Kumar A, Meena RS (2015b) Issues and challenges about sus-
tainable agriculture production for management of natural resources to sustain soil fertility and
health. J Clean Prod 107:793–794
Yadav BK, Sidhu AS (2016) Dynamics of potassium and their bioavailability for plant nutrition. In:
Meena VS, Maurya BR, Verma JP, Meena RS (eds) Potassium solubilizing microorganisms for
sustainable agriculture. Springer, New Delhi, pp 187–201. doi:10.1007/978-81-322-2776-2_14
Yasin M, Munir I, Faisal M (2016) Can Bacillus spp. enhance K+ uptake in crop species. In:
Meena VS, Maurya BR, Verma JP, Meena RS (eds) Potassium solubilizing microorganisms for
sustainable agriculture. Springer, New Delhi, pp 163–170. doi:10.1007/978-81-322-2776-2_12
Zahedi H (2016) Growth-promoting effect of potassium-solubilizing microorganisms on
some crop species. In: Meena VS, Maurya BR, Verma JP, Meena RS (eds) Potassium sol-
ubilizing microorganisms for sustainable agriculture. Springer, New Delhi, pp 31–42.
doi:10.1007/978-81-322-2776-2_3
The Role of Soil Microbes in Crop
Biofortification 16
Darren Heenan Daly, Siva L.S. Velivelli,
and Barbara Doyle Prestwich
Abstract
Agronomic practices across the planet are becoming largely unsustainable in
their current forms. With a growing population expected to reach ~9 billion by
the year 2050, more sustainable ways to produce the world’s main crops are
needed. The main focus of current agronomic practices, especially in the case of
cereal crops, is increased grain number and weight sometimes at the expense of
nutritional content leading, in some instances, to micronutrient deficiencies.
Micronutrient deficiencies are often termed hidden hunger, giving the false
appearance that an individual is consuming sufficient amounts of nutrients. To
counteract this problem, it is crucial that a sustainable solution to increase
micronutrient concentration in a diverse range of crops is found. Plant growth-
promoting microbes (PGPM) represents a sustainable solution to this problem.
These PGPM can be divided into two main groups: plant growth-promoting
rhizobacteria (PGPR) and plant growth-promoting fungi (PGPF). These
microbes are capable of increasing micronutrient concentrations in many crops
worldwide. This chapter will focus on the use of these microbes to increase
micronutrient content, in particular selenium, iron and zinc, using studies
conducted over the last two decades right up to the present day, revealing
how plant-microbe interactions and our ever-growing knowledge of these inter-
actions can aid in the micronutrient biofortification of crops in a sustainable and
environmentally friendly way.
Keywords
Plant growth-promoting microbes • Sustainability • Micronutrients • Biofortification
16.1 Introduction
16.2 T
he Application of Plant Growth-Promoting
Rhizobacteria for the Biofortification of Crops
The rhizosphere, an important interface between plant roots and the soil, can con-
tribute to sustainable agriculture when the interaction between plants and beneficial
bacteria is exploited. It has been ~35 years since Kloepper first described the role of
plant growth-promoting rhizobacteria (PGPR) in plant growth and defence
(Kloepper et al. 1980). PGPR when associated with rhizosphere/plant roots play a
major role in the direct or indirect promotion of plant growth. The direct plant
growth promoter mechanisms, biofertilization and phytostimulation, are two exam-
ples of methods that simultaneously minimize the use of chemical fertilizers and
promote plant growth, and bacteria with both biocontrol and biofertilization/phyto-
stimulation properties offer advantage to plants in terms of both enhanced nutrient
supply and control of disease (Adesemoye and Kloepper 2009; Lugtenberg and
Kamilova 2009; Glick 2012; Velivelli et al. 2014).
The recent work in the area of plant-microbe interactions has focused on the
biofortification of staple crops using these PGPR, showing significant results. The
WHO has identified micronutrients which are essential to the correct functioning of
the human body, i.e. selenium (Se), iron (Fe) and zinc (Zn), and these constitute a
significant portion of the current research on PGPR-mediated biofortification
(Priyadharsini and Muthukumar 2016; Kumar et al. 2017; Meena et al. 2015a, b, f;
Raghavendra et al. 2016; Zahedi 2016; Dominguez-Nunez et al. 2016; Dotaniya
et al. 2016). These three micronutrients will be discussed further in relation to the
biofortification of crops using PGPR.
16.2.1 Selenium
such as anti-oxidation and prevention of certain cancers, e.g. liver cancer (Carlson
et al. 2012) and colorectal cancer (Meplan and Hesketh 2012). Selenoproteins, i.e.
proteins that contain a selenocysteine amino acid residue, facilitate adequate DNA
synthesis, protection against diseases such as HIV, reproduction and thyroid hor-
mone metabolism (Sunde 2012; Hatfield et al. 2014). A delicate balance occurs in
humans between having too much and too little Se in the diet. Se levels that are
too high can lead to selenotoxicity, resulting in symptoms ranging from nervous
system disorders to nail and hair loss (Li et al. 2012), and selenium levels that are
too low can lead to inadequate levels of protection against radioactive damage to
DNA, which can result in micronuclei formation and potentially cancer (Baliga
et al. 2008).
mustard plants, the Se volatilization rate from selenite was four times higher than
that of axenic control plants, and plants accumulated more Se in their tissues when
compared to axenic controls. Both BJ2 and BJ15 rhizobacterial strains increased Se
concentrations in shoots and roots 1.4-fold and fivefold, respectively. As with many
PGPR species, these isolates were thought to increase Se uptake via enhanced root
hair production. In addition, seedlings with PGPR had a significantly higher Se tis-
sue concentration of 1.3–2 times greater than axenic plants across the spectrum of
Se concentrations tested (de Souza et al. 1999).
Acuña et al. (2013) investigated ash-derived volcanic andisol soil in Southern
Chile, which is characteristically low in Se content, for the presence of PGPR with
the ability to supply increased Se to host plants. Inoculation with two isolates,
namely, Pseudomonas sp. R8 and Stenotrophomonas sp. B19, in the rhizosphere of
wheat resulted in higher Se content in roots and leaves compared to uninoculated
controls. When plants were inoculated with B19 in soils where the Se concentration
was 5–10 mM, a significantly higher Se content was observed in wheat root and leaf
tissues compared to plants grown in soil containing 2 mM Se (Acuña et al. 2013).
In the same region, Durán et al. (2014) isolated endophytic bacteria from
Se-supplemented wheat that shows potential for plant growth promotion, biofortifi-
cation and biocontrol in wheat cultivation (Saha et al. 2016c; Verma et al. 2014,
2015b; Meena et al. 2014a, 2015e; Teotia et al. 2016).
Endophytic bacterial strains were isolated from both uninoculated control and
Se-supplemented wheat plants from the genera Bacillus, Paenibacillus, Klebsiella,
Enterobacter and Acinetobacter and 15 strains selected according to tolerance of
high Se levels (~50 mM). A number of these endophytes are highly tolerant to Se,
ranging from 60 to 180 mM, and thus have the potential to be used as biofortifica-
tion agents in areas of the world with low soil Se concentrations. Furthermore, some
of these isolates displayed the ability to act as biocontrol agents for the endemic
soilborne fungal pathogen Gaeumannomyces graminis var. tritici that causes take-
all disease, one of the most severe wheat diseases in Southern Chile. Isolates from
the genera Acinetobacter (strain E6.2), Bacillus (strain E8.1), Bacillus and Klebsiella
(strains E5 and E1) inhibited the growth of G. graminis var. tritici mycelia by
~100%, 50% and 30%, respectively, in vitro; this adds to the potential of PGPR for
biofortification agents that not only increase Se concentration in the plant but also
act as sustainable crop protectants (Durán et al. 2014).
Two studies by Yasin et al. (2015) investigated the potential for Se biofortifica-
tion in crop plants. First, an experiment with B. juncea to investigate the potential of
two Se-tolerant rhizobacterial consortia to increase plant Se uptake using naturally
seleniferous soil containing ~8 mg Se kg−1 was carried out with both rhizobacterial
consortia consisting of four to five strains each, designated G1 and G2. B. juncea
accumulated Se to 358 mg kg−1 in seeds, 711 mg kg−1 in leaf dry weight (DW) and
276 mg kg−1 in pod husks. Inoculation of G1 had a positive effect on plant growth
compared to controls but did not accumulate Se with a significant difference com-
pared to the control. G1-incoulated plants produced 40–45% more leaf and seed
biomass compared to controls however; therefore, these plants were more effective
in extracting Se from the soil compared to control plants. G2-inoculated plants had
338 D.H. Daly et al.
16.2.2 Iron
Se
Se Se Sample
Se Se
Se
Se
a b
Cultivate
Inoculate
Se
Se
Fig. 16.1 Rationale for selection of PGPR involved in biofortification taking Se as an example.
(a) Se-enriched soils. (b) Isolation of bacteria from (a) on Se-supplemented growth media to iden-
tify Se-tolerant bacteria. (c) Inoculation of PGPR from Se-enriched soils to Se-deficient soils to aid
Se biofortification in plants
Table 16.1 Recommended daily dietary intake of Se, Zn and Fe, WHO (2006)
Se Zn Fe
Category mg
Infants (1–3 years) 17 2.4–8.3 3.9–11.6
Children (4–6 years) 22 2.9–9.6 4.2–12.6
Adult female 26 3–9.8 19.6–58.8
Adult male 34 4.2–14 9.1–27.4
Pregnant women 28 4.2–14 >50
Lactating women <3 months 35 5.8–19 10–30
Values for Zn and Fe run from low to high as bioavailability increases in the diet. A diet of low
bioavailability is defined as a diet low in vitamin C and animal protein; the opposite is true for a
diet with high bioavailability. Socioeconomic and dietary factors play a major role in the potential
uptake of micronutrients in the human body
1999; Nuberger et al. 2016). So, as for all (bio)fortification strategies, the relative
benefits and risks must be balanced (Meena et al. 2013c, 2014b, 2015d; Singh et al.
2015, 2016).
required for induction of ferric reductase FRO2 and the IRT1 iron transporter to
acquire Fe in the plant (Zhang et al. 2009).
A group of major microbial-derived plant growth-promoting (PGP) hormones
called auxins have been implicated in enhanced Fe uptake. The ratio of auxin-
producing microorganisms in soil that contains phenolic root exudates of Fe-deficient
red clover (Trifolium pratense L.) is higher than that observed in phenolic-free soil
solution controls, thus indicating that Fe deficiency in plants may contribute to the
abundance of auxin-producing microbes in the rhizosphere. Not only the increase of
auxin-producing microbes was observed, but the plant phenolics also inhibited the
growth of the majority of microbes, suggesting iron-deficient plants can select for
organisms which have the ability to produce auxin and siderophores, ~71% and
86% of the total microbial population, respectively (Jin et al. 2006, 2008). Exogenous
application of the major auxin, indole-3-acetic acid (IAA), enhances Fe deficiency-
induced reduction of ferric Fe and expression of FRO2 and IRT1, which are critical
for enhanced Fe uptake in the plant and growth of lateral roots and root hairs to
provide increased surface area for Fe uptake (Jin et al. 2008; Chen et al. 2010; Wu
et al. 2012). IAA also contributes extra nutrients for rhizobacterial growth by modu-
lating the quantity of root exudation via an increase of porosity in the cell wall of
plants (Glick 2012).
Utilizing two of the most important cereal crops, rice (Oryza sativa L.) and
wheat (Triticum aestivum) in a rice-wheat cropping system, Rana et al. (2015) dem-
onstrated that PGPR and cyanobacteria led to both higher macro- and micronutrient
concentration in cereal grain. In rice, bacterial-cyanobacterial consortia increased
Fe concentration from ~4 to 14% over the fertilizer-only control. While bacterial-
cyanobacterial consortia had a significant Fe increase over uninoculated controls,
isolate Providencia sp. PW5 alone increased the Fe concentration by ~45% com-
pared to the fertilizer-only control in wheat grains. Adak et al. (2016) also observed
increased acquisition of Fe through cyanobacteria inoculation in rice of ~13 to 46%
compared to uninoculated controls.
Recent publications from India on the biofortification of chickpea (Cicer arieti-
num L.), a major pulse crop grown under semiarid conditions, have highlighted the
potential for Fe biofortification in chickpea. Nineteen Acinetobacter isolates statis-
tically significantly increased mineral content in PGPR-inoculated chickpeas com-
pared to uninoculated controls; Fe in plants varied from ~10 to 38% higher (Sathya
et al. 2016). All 19 isolates increased Fe in the plant within the range of ~10 to 38%.
Gopalakrishnan et al. (2016) also recorded increases in Fe concentration following
PGPR treatment. Fe concentration in chickpea grain was increased up to 18% with
isolates E. ludwigii SRI-229 and P. monteilii SRI-360, the two strains displaying the
greatest performance over uninoculated controls (Gopalakrishnan et al. 2016). It is
important to note that postharvest processing and cooking can affect micronutrient
concentrations in the grain. Both Saytha et al. (2016) and Gopalakrishnan et al.
(2016) reported that Fe concentration can be affected by cooking and can reduce
from 5 to 30% and in some cases gain ~21% (Sathya et al. 2016; Gopalakrishnan
et al. 2016).
342 D.H. Daly et al.
16.2.3 Zinc
Table 16.2 A selections of microbes which have been shown to contribute to the process of
micronutrient biofortification
Microbe Micronutrient Reference
Bacillus subtilis (GB 03) Fe Zhang et al. (2009)
Pseudomonas sp. R8 Se Acuña et al. (2013)
Stenotrophomonas sp. B19 Se Acuña et al. (2013)
Providencia sp. PW5 Zn, Fe, Mn, Cu Rana et al. (2012)
Pseudomonas sp. Z5 Zn Yasmin (2011)
Flavobacterium sp. Zn He et al. (2010)
Anabaena sp. Fe Manjunath et al. (2016)
Azotobacter sp. Fe, Zn Manjunath et al. (2016)
Calothrix sp. Zn Manjunath et al. (2016)
Pseudomonas fluorescens C7 Fe Vansuyt et al. (2007)
16.3 T
he Application of Arbuscular Mycorrhizal Fungi (AMF)
to Biofortification in Crops
Most plants, including all major food crops, form a symbiotic below-ground asso-
ciation with AMF that supplies the host plant with water and mineral nutrients, such
as phosphorus (Senés-Guerrero et al. 2014). Plants colonized by AMF can effec-
tively acquire nutrients from a larger soil volume, beyond the nutrition depletion
zone to where the plant roots cannot extend. Thus, AMF helps the plant to grow
better and increase its productivity. In return, the host plant provides AMF with
carbohydrates required to complete its life cycle (Zhang et al. 2015).
The AMF acquire essential nutrients such as P, Zn, Cu, Fe, N and K from the soil,
and thus AMF contact with roots provides the plant with access to essential nutri-
tional elements, leading to solubilization, mobilization and uptake of the essential
nutrients needed by plants for their growth (Clark and Zeto 2000; Kumar et al.
2016a). Plants colonized with AMF have significantly greater zinc concentrations in
all tissue types compared to non-mycorrhizal plants, although the effect is primarily
344 D.H. Daly et al.
Endophytic
Space
ACC
IAA b
Siderophore
a
Zn Root Exudates
Fe Se
c Se
Se Se
d
Fig. 16.2 Schematic representation of the reciprocal relationship between PGPR and the host
plant; (a) represents the plant rhizospheric space, (b) represents beneficial PGPR. The plant pro-
vides root exudates which are consumed by bacteria as a carbon source, and endophytic PGPR
have the option to potentially colonize the plant and avoid adverse environmental conditions. (c)
Represents the increased uptake of micronutrients in the rhizosphere where PGPR activity leads to
micronutrient biofortification of the plant. (d) Represents micronutrients in the soil which would
be either unavailable or diminished in available supply to the plant without PGPR. Based on
numerous PGP activities, PGPR can increase the translocation and concentration of key micronu-
trients which would otherwise be unavailable to the plant. Especially in the case of siderophore-
producing PGPR, which can have a beneficial effect on Fe sequestration from the soil and
subsequent transfer to the plant. In return, the plant can supply the PGPR with a carbon source in
the form of root exudates and where there are endophytic PGPR present; the plant endosphere can
act as a suitable environment for endophytic proliferation
due to soil property differences (Lehmann et al. 2014). Improved total uptake of
N-P-K was observed in both shoots and roots of Plantago minuta under variable soil
water conditions, when inoculated with AMF (Shi et al. 2015). Under field condi-
tions, inoculation of AMF in the soil of chickpea plants increased plant biomass and
yield, and also increased the nutritional value of the legume, leading to Fe and Zn
biofortification (Pellegrino and Bedini 2014).
In addition, AMF increases positive interactions with rhizobacteria, including
beneficial rhizobacteria known to induce spore germination of AMF and root colo-
nization directly. AMF hyphal exudates influence surrounding microbial communi-
ties and may drive beneficial bacterial-plant interactions and consequently improve
plant fitness and productivity (Scheublin et al. 2010; Cruz and Ishii 2011; Qin et al.
16 The Role of Soil Microbes in Crop Biofortification 345
The threats to global food security have been highlighted by many authors to date
(Tai et al. 2014; Sundström et al. 2014). National governments through the interna-
tional forum of the United Nations have agreed on 17 goals to be achieved by 2030
(Sustainable Development Goals project), amongst them the eradication of world
hunger and improved nutrition (FAO 2016). A number of these goals, particularly in
relation to food security, can be addressed by crop biofortification. Many successful
strategies for biofortifying crops are detailed in the peer-reviewed literature with the
most famous biofortification project concerning the expression of provitamin A in
Golden Rice which was based on a genetic engineering strategy. Even though the
Golden Rice project was awarded the Patents for Humanity prize by the United
States Patent and Trademark Office (USPTO) in 2015 in recognition of this lifesav-
ing technology and the humanitarian nature of the project, the product is still not
available on the market due to public opposition, in the main led by Greenpeace.
This year (2016), 121 Nobel Laureates called on Greenpeace to end its opposition
to this lifesaving technology (Support Precision Agriculture 2016).
Aside from genetic modification, other successful strategies including conven-
tional breeding strategies have been utilized for crop biofortification. This year
(2016), the World Food Prize was awarded to a group of scientists in recognition of
their role in crop biofortification using breeding strategies to increase provitamin A
in sweet potato by incorporating germplasm from the Andean region in South
America to generate orange-fleshed sweet potatoes suitable for Africa (ISAAA
2016). This breeding strategy is reliant on natural diversity in the germplasm of a
crop for the production of a particular vitamin or micronutrient.
In the absence of natural germplasm diversity, other strategies for crop bioforti-
fication can be utilized including foliar application of micronutrients and/or the
exploitation of the natural microbial diversity in soils, i.e. plant growth-promoting
microbes (PGPM) most especially bacteria and mycorrhizal fungi. Being environ-
mentally safe and cost-effective are major advantages to the proliferation of PGPM
throughout agriculture. Indeed this can be seen by the vast number of commercially
available PGPM products released in recent years with up to 4% of the global
biocide share being directly related to microbial volatile organic compounds
(mVOCs) isolated and synthesized from PGPM (Glare et al. 2012; Wilson et al.
2013; Velivelli et al., 2014; Kanchiswamy et al. 2016), and this trend is set to
continue. A historic problem in the biofortification of crops using foliar and soil
application of micronutrients particularly in relation to Fe has been the low uptake
of the micronutrient post-application. This has been highlighted recently by
Kromann et al. (2016) with regard to Fe and Zn biofortification of Andean potatoes.
It was found that both foliar and soil application of Zn fertilizers led to a ~three- and
twofold increase, respectively, in tuber Zn concentration. However, there was no
increase in Fe tuber concentration post-application. In this instance, it can be
suggested that application of Fe fertilizer in conjunction with a PGPM capable of
increasing Fe translocation and concentration in the tuber could be tested. The study
16 The Role of Soil Microbes in Crop Biofortification 347
also highlights the need to understand soil parameters within diverse environments,
as these can have a significant effect on crop biofortification (Kromann et al. 2016).
Regulatory mechanisms in the plant-microbe micro-/macronutrient cycling path-
ways and their reciprocal interactions need to be further elucidated to determine the
best course of action for formulation of appropriate treatments to specific soil/cli-
mate/temperature/crop scenarios. Indeed, this can be further seen in the observation
that nitrogen status of the plant can have a positive correlation on the content and
translocation of Fe and Zn in plants (Kutman et al. 2010, 2011), and many plant
growth-promoting microbes are capable of fixing atmospheric nitrogen (Prasanna
et al. 2012). Genome mining of selected PGPR or wild relative/stress-tolerant culti-
vars of major crops may reveal genes involved in nutrient cycling pathways that,
through GM technology, could be incorporated into agriculturally important crops
to optimize delivery of micronutrients to deficient populations in the developing
world.
Using genomics, identifying changes in gene expression during microbial inter-
actions is critical to understanding the role of microbes in biofortification. In par-
ticular, more attention needs to be focused on the following research questions: how
microbes influence the nutritional content of economically important crops; identi-
fying appropriate microbial strains; exploiting synergistic microbial activity, e.g.
between AMF and other beneficial bacteria; and assessing constant field efficacy
under different environmental regimes, to ensuring adequate uptake by plants and
improving crop quality. Supplying essential nutrients using beneficial microbes
naturally found in soils across the planet, such as AMF and PGPR, will help to alle-
viate the problem of hidden hunger and provide a promising sustainable agricultural
strategy for improving current crop micronutrient content and in developing future
biofortified crops.
Acknowledgements The authors would like to acknowledge the contribution of the Crawford-
Hayes fund at the University College Cork, Ireland, for its contribution to the writing of this
chapter.
References
Acuña JJ, Jorquera MA, Barra PJ, Crowley DE, de la Luz MM (2013) Selenobacteria selected
from the rhizosphere as a potential tool for Se biofortification of wheat crops. Biol Fertil Soils
49:175–185. doi:10.1007/s00374-012-0705-2
Adak A, Prasanna R, Babu S, Bidyarani N, Verma S, Pal M, Shivay YS, Nain L (2016) Micronutrient
enrichment mediated by plant-microbe interactions and rice cultivation practices. J Plant Nutr
39:1216–1232. http://dx.doi.org/10.1080/01904167.2016.1148723
Adesemoye AO, Kloepper JW (2009) Plant-microbes interactions in enhanced fertilizer-use effi-
ciency. Appl Microbiol Biotechnol 85:1–12
Ahmad M, Nadeem SM, Naveed M, Zahir ZA (2016) Potassium-solubilizing bacteria and their
application in agriculture. In: Meena VS, Maurya BR, Verma JP, Meena RS (eds) Potassium
solubilizing microorganisms for sustainable agriculture. Springer, India, pp 293–313.
doi:10.1007/978-81-322-2776-2_21
348 D.H. Daly et al.
Amanullah SA, Iqbal A, Fahad S (2016) Foliar phosphorus and zinc application improve growth
and productivity of maize (Zea mays L.) under moisture stress conditions in semi-arid climates.
J Microb Biochem Technol 8:433–439. doi:10.4172/1948-5948.1000321
Bahadur I, Meena VS, Kumar S (2014) Importance and application of potassic biofertilizer in
Indian agriculture. Int Res J Biol Sci 3:80–85
Bahadur I, Maurya BR, Kumar A, Meena VS, Raghuwanshi R (2016a) Towards the soil sustain-
ability and potassium-solubilizing microorganisms. In: Meena VS, Maurya BR, Verma JP,
Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture. Springer,
India, pp 225–266. doi:10.1007/978-81-322-2776-2_18
Bahadur I, Maurya BR, Meena VS, Saha M, Kumar A, Aeron A (2016b) Mineral release dynamics
of tricalcium phosphate and waste muscovite by mineral-solubilizing rhizobacteria isolated
from indo-gangetic plain of India. Geomicrobiol J. doi:10.1080/01490451.2016.1219431
Baliga MS, Diwadkar-Navsariwala V, Koh T, Fayad R, Fantuzzi G, Diamond AM (2008)
Selenoprotein deficiency enhances radiation-induced micronuclei formation. Mol Nutr Food
Res 52(11):1300–1304. doi:10.1002/mnfr.200800020
Beard JL (2008) Why iron deficiency is important in infant development. J Nutr 138:2534–2536
Bell PF, Parker DR, Page AL (1992) Contrasting selenite sulfate interactions in se accumulating
and nonaccumulating plant species. Soil Sci Soc Am 56:1818–1824. doi:10.1236/sssaj1992.0
3615995005600060028x
Berutti A, Lumini E, Balestrini R, Bianciotto V (2016) Arbuscular mycorrhizal fungi as natu-
ral biofertilizers: let’s benefit from past successes. Front Microbiol 6:1559. doi:10.3389/
fmicb.2015.01559
Bhutta ZA (2008) Micronutrient needs of malnourished children. Curr Opin Clin Nutr Metab Care
11:309–314
Black RE, Lindsay HA, Bhutta ZA, Caulfield LE, De Onnis M, Ezzati M, Mathers C, Rivera
J (2008) Maternal and child undernutrition: global and regional exposures and health conse-
quences. Lancet 371:243–260
Boukhalfa H, Lack JG, Reilly SD, Hersman L, Neu MP (2003) Siderophore production and facili-
tated uptake of iron and plutonium in P. putida. No. LA-UR-03-0913. Los Alamos National
Laboratory
Boyer LR, Brain P, Xu XM, Jeffries P (2015) Inoculation of drought-stressed strawberry with a
mixed inoculum of two arbuscular mycorrhizal fungi: effects on population dynamics of fungal
species in roots and consequential plant tolerance to water deficiency. Mycorrhiza 25(3):215–
227. doi:10.1007/s00572-014-0603-6
Broadley MR, White PJ, Bryson RJ, Meacham MC, Bowen HC, Johnson SE, Hawkesford MJ,
McGrath SP, Zhao F-J, Breward N, Harriman M, Tucker M (2006) Biofortification of UK food
crops with selenium. Proc Nutr Soc 65:169–181
Broadley MR, White PJ, Zelko I, Lux A (2007) Zinc in plants. New Phytol 173(4):677–702
Bulgarelli D, Schlaeppi K, Spaepen S, van Themaat EVL, Schulze-Lefert P (2013) Structure and
functions of the bacterial microbiota of plants. Annu Rev Plant Biol 64:807–838. doi:10.1146/
annurev-arplant- 050312-120106
Cakmak I (2008) Enrichment of cereal grains with zinc: agronomic or genetic bio-fortification?
Plant Soil 302:1–17
Carlson BA, Yoo MH, Tobe R, Mueller C, Naranjo-Suarez S, Hoffmann VJ, Gladyshev VN, Hatfield
DL (2012) Thioredoxin reductase 1 protects against chemically induced hepatocarcinogenesis
via control of cellular redox homeostasis. Ox J Carcin 33(9):1806–1803. doi:10.1093/carcin/
bgs230
Chen WW, Yang JL, Qin C, Jin CW, Mo JH, Ye T, Zheng SJ (2010) Nitric oxide acts down-
stream of auxin to trigger root ferric-chelate reductase activity in response to iron deficiency in
Arabidopsis. Plant Physiol 154:810–819
Clark RB, Zeto SK (2000) Mineral acquisition by arbuscular mycorrhizal plants. J Plant Nutr
23(7):867–902. doi:10.1080/01904160009382068
16 The Role of Soil Microbes in Crop Biofortification 349
Crowley DA (2006) Microbial siderophores in the plant rhizosphere. In: Barton LL, Abadia J (eds)
Iron nutrition in plants and rhizospheric microorganisms. Springer, Netherlands, pp 169–189
Cruz AF, Ishii T (2011) Arbuscular mycorrhizal fungal spores host bacteria that affect nutrient
biodynamics and biocontrol of soil-borne plant pathogens. Biol Open 1(1):52–57. doi:10.1242/
bio.2011014
Das I, Pradhan M (2016) Potassium-solubilizing microorganisms and their role in enhancing
soil fertility and health. In: Meena VS, Maurya BR, Verma JP, Meena RS (eds) Potassium
solubilizing microorganisms for sustainable agriculture. Springer, India, pp 281–291.
doi:10.1007/978-81-322-2776-2_20
De Souza M, Chu D, Zhao M, Zayed AM, Ruzin SE, Schichnes D, Terry N (1999) Rhizosphere
bacteria enhance selenium accumulation and volatilization by Indian mustard. Plant Physiol
119:565–573
Dominguez-Nunez JA, Benito B, Berrocal-Lobo M, Albanesi A (2016) Mycorrhizal fungi: role
in the solubilization of potassium. In: Meena VS, Maurya BR, Verma JP, Meena RS (eds)
Potassium solubilizing microorganisms for sustainable agriculture. Springer, India, pp 77–98.
doi:10.1007/978-81-322-2776-2_6
Dotaniya ML, Meena VD, Basak BB, Meena RS (2016) Potassium uptake by crops as well
as microorganisms. In: Meena VS, Maurya BR, Verma JP, Meena RS (eds) Potassium
solubilizing microorganisms for sustainable agriculture. Springer, India, pp 267–280.
doi:10.1007/978-81-322-2776-2_19
Durán P, Acuña JJ, Jorquera MA, Azcón R, Borie F, Cornejo P, Mora ML (2013) Enhanced sele-
nium content in wheat grain by co-inoculation of selenobacteria and arbuscular mycorrhizal
fungi: a preliminary study as a potential Se biofortification strategy. J Cereal Sci 57(3):275–
280. doi: http://dx.doi.org/10.1016/j.jcs.2012.11.012
Durán P, Acuña JJ, Jorquera MA, Azcón R, Paredes C, Rengel Z, de la Luz MM (2014) Endophytic
bacteria from selenium-supplemented wheat plants could be useful for plant-growth promo-
tion, biofortification and Gaeumannomyces graminis biocontrol in wheat production. Biol
Fertil Soils 50:983–990
Ellis DR, Salt DE (2003) Plants, selenium and human health. Curr Opin Plant Biol 6:273–279.
doi:10.1016/S1369-5266(03)00030-X
FAO (2016) The state of food and agriculture. http://www.fao.org/publications/sofa/sofa2016/en/
Gallou A, Lucero Mosquera HP, Cranenbrouck S, Suárez JP, Declerck S (2011) Mycorrhiza
induced resistance in potato plantlets challenged by Phytophthora infestans. Physiol Mol Plant
Pathol 76(1):20–26. doi:http://dx.doi.org/10.1016/j.pmpp.2011.06.005
Gamalero E, Glick B (2011) Mechanisms used by plant growth-promoting bacteria. In: Maheshwari
DK (ed) Bacteria in agrobiology: plant nutrient management. Springer, Berlin, pp 17–46
Ganther HE, Levander OA, Saumann CA (1966) Dietary control of selenium volatilisation in the
rat. J Nutr 88:55–60
Glare T, Caradus J, Gelernter W, Jackson T, Keyhani N, Köhl J, Marrone P, Morin L, Stewart
A (2012) Have biopesticides come of age? Trends Biotechnol 30(5):250–258. doi:10.1016/j.
tibtech.2012.01.003
Glick BR (2012) Plant growth-promoting bacteria: mechanisms and applications. Scientifica
2012:1–15
Golden Rice (2016). www.goldenrice.org
Gopalakrishnan S, Vadlamundi S, Samineni S, Sameer Kumar CV (2016) Plant growth-promotion
of chickpea and pigeon pea through inoculation of biocontrol potential bacteria, isolated from
organic soils. Springer Plus 5:1882. doi:10.1186/s40064-016-3590-6
Hafeez FY, Hameed S, Zaidi AH, Malik KA (2002) Biofertiliser for sustainable agriculture. In:
Azam F, Iqbal MM, Inayatullah C, Malik KA (eds) Technologies for sustainable agriculture.
NIAB, Faisalabad, pp 67–74
Harvest Plus (2016). http://www.harvestplus.org/biofortification-nutrition-revolution-now
Hatfield DL, Tsuji PA, Carlson BA, Gladyshev VN (2014) Selenium and selenocysteine:
roles in cancer, health and development. Trends Biochem Sci 39:112–120. doi:10.1016/j.
tibs.2013.12.007
350 D.H. Daly et al.
Hawkesford MJ, Zhao F-J (2007) Strategies for increasing the selenium content of wheat. J Cereal
Sci 46:282–292
He X, Nara K (2007) Element biofortification: can mycorrhizas potentially offer a more effec-
tive and sustainable pathway to curb human malnutrition? Trends Plant Sci 12(8):331–333.
doi:10.1016/j.tplants.2007.06.008
He CQ, Tan G, Liang X, Du W, Chen YL, Zhi GY, Zhu Y (2010) Effect of Zn-tolerant bacterial
strain on growth and Zn accumulation in Orychophragmus violaceus. Appl Soil Ecol 44:1–5
International Rice Research Institute (2016). http://irri.org/golden-rice/faqs
ISAAA (2016). http://www.isaaa.org/kc/cropbiotechupdate/article/default.asp?ID=14884
Jaiswal DK, Verma JP, Prakash S, Meena VS, Meena RS (2016) Potassium as an important plant
nutrient in sustainable agriculture: a state of the art. In: Meena VS, Maurya BR, Verma JP,
Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture. Springer,
India, pp 21–29. doi:10.1007/978-81-322-2776-2_2
Jat LK, Singh YV, Meena SK, Meena SK, Parihar M, Jatav HS, Meena RK, Meena VS (2015)
Does integrated nutrient management enhance agricultural productivity? J Pure Appl Microbiol
9(2):1211–1221
Jha Y, Subramanian RB (2016) Regulation of plant physiology and antioxidant enzymes for alle-
viating salinity stress by potassium-mobilizing bacteria. In: Meena VS, Maurya BR, Verma JP,
Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture. Springer,
India, pp 149–162. doi:10.1007/978-81-322-2776-2_11
Jin CW, He YF, Tang CX, Wu P, Zheng SJ (2006) Mechanisms of microbially enhanced iron
uptake in red clover. Plant Cell Environ 29:888–897
Jin CW, You GY, Zheng SJ (2008) The iron-deficiency induced phenolics secretion plays multiple
important roles in plant iron acquisition underground. Plant Signal Behav 3:60–61
Kanchiswamy CN, Malnoy M, Maffei ME (2016) Bioprospecting bacterial and fungal volatiles
for sustainable agriculture. Trends Plant Sci 20(4):206–211. doi:10.1016/j.tplants.2015.01.004
Khush GS, Lee S, Cho J-I, Jeon J-S (2012) Biofortification of crops for reducing malnutrition.
Plant Biotechnol Reports 6(3):195–202. doi:10.1007/s11816-012-0216-5
King JC (2006) Zinc. In: Shills ME, Shike M (eds) Modern nutrition in health and disease, 10th
edn. Lippincott Williams and Wilkins, Philadelphia, pp 271–285
King JC (2011) Zinc: an essential but elusive nutrient. Am J Clin Nutr 94(suppl):697S–684S
Kloepper JW, Schroth MN, Miller TD (1980) Effects of rhizosphere colonization by plant growth-
promoting rhizobacteria on potato plant development and yield. Phytopathology 70:1078–1082
Krishnakumar S, Balakrishnan N, Muthukrisnan R, Kumar SR (2013) Myth and mystery of soil
mycorrhiza: a review. Af J Ag Res 8(38):4706–4717. doi:10.5897/AJAR2013.7490
Kromann P, Valverde F, Alvardo S, Vélez R, Pisuña J, Potosí B, Taipe A, Caballero D, Cabezas A,
Devaux A (2016) Can Andean potatoes be agronomically biofortified with iron and zinc fertil-
isers? Plant Soil. doi:10.1007/s11104-016-3065-0
Kumar A, Bahadur I, Maurya BR, Raghuwanshi R, Meena VS, Singh DK, Dixit J (2015)
Does a plant growth-promoting rhizobacteria enhance agricultural sustainability? J Pure
ApplMicrobiol 9:715–724
Kumar A, Choudhary AK, Pooniya V, Suri VK, Singh U (2016a) Soil factors associated with
micronutrient acquisition in crops- biofortification perspective. In: Singh U, Praharaj
SC, Singh SS, Singh PN (eds) Biofortification of food crops. Springer, India, pp 159–176.
doi:10.1007/978–81–322-2716-8_13
Kumar A, Meena R, Meena VS, Bisht JK, Pattanayak A (2016b) Towards the stress management
and environmental sustainability. J Clean Prod 137:821–822
Kumar A, Patel JS, Bahadur I, Meena VS (2016c) The molecular mechanisms of KSMs for
enhancement of crop production under organic farming. In: Meena VS, Maurya BR, Verma JP,
Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture. Springer,
India, pp 61–75. doi:10.1007/978-81-322-2776-2_5
Kumar A, Maurya BR, Raghuwanshi R, Meena VS, Islam MT (2017) Co-inoculation with
enterobacter and rhizobacteria on yield and nutrient uptake by wheat (Triticum aestivum L.)
16 The Role of Soil Microbes in Crop Biofortification 351
in the alluvial soil under indo-gangetic plain of India. J Plant Growth Regul. d oi:10.1007/
s00344-016-9663-5
Kutman UB, Yildiz B, Ozturk L, Cakmak I (2010) Biofortification of durum wheat with zinc
through soil and foliar application of nitrogen. Cereal Chem 87:1–9
Kutman UB, Yildiz B, Cakmak I (2011) Improved nitrogen status enhances zinc and iron concen-
trations both in the whole grain and the endosperm fraction of wheat. J Cereal Sci 53:118–125
Lehmann A, Veresoglou SD, Leifheit EF, Rillig MC (2014) Arbuscular mycorrhizal influence
on zinc nutrition in crop plants – a meta-analysis. Soil Biol Biochem 69:123–131. doi:http://
dx.doi.org/10.1016/j.soilbio.2013.11.001
Li SH, Xiao TF, Zheng BS (2012) Medical geology of arsenic, selenium and thallium in China. Sci
Total Environ 421–422:31–40. doi:10.1016/j. scitotenv.201102.040
Lugtenberg B, Kamilova F (2009) Plant growth-promoting rhizobacteria. Annu Rev Microbiol
63:541–556
Mäder P, Kaiser F, Adholeya A, Singh R, Uppal HS, Sharma AK, Srivastava R, Sahai V, Aragno
M, Wiemken A, Johri BN, Fried PM (2011) Inoculation of root microorganisms for sustain-
able wheat–rice and wheat–black gram rotations in India. Soil Biol Biochem 43(3):609–619.
doi:http://dx.doi.org/10.1016/j.soilbio.2010.11.031
Manjunath M, Kanchan A, Ranjan K, Venkatachalam S, Prasanna R, Ramakrishnan B, Hossain
F, Nain L, Shivay YS, Rai AB, Singh B (2016) Beneficial cyanobacteria and eubacteria syn-
ergistically enhance bioavailability of soil nutrients and yield of Okra. Heliyon http://dx.doi.
org/10.1016/j.heliyon.2016.c00066
Masalha J, Kosegarten H, Elmaci Ö, Mengel K (2000) The central role of microbial activity for
iron acquisition in maize and sunflower. Biol Fertil Soils 30:433–439
Masood S, Bano A (2016) Mechanism of potassium solubilization in the agricultural soils by
the help of soil microorganisms. In: Meena VS, Maurya BR, Verma JP, Meena RS (eds)
Potassium solubilizing microorganisms for sustainable agriculture. Springer, India, pp 137–
147. doi:10.1007/978-81-322-2776-2_10
Maurer B, Keller-Scheirlin W (1968) Ferribactin, a siderophore from Pseudomonas fluorescens
Migula 61. Arch Mirobiol 60:326–339
Maurya BR, Meena VS, Meena OP (2014) Influence of inceptisol and alfisol’s potassium solubiliz-
ing bacteria (KSB) isolates on release of K from waste mica. Vegetos 27:181–187
Mayer JE, Pfeiffer WH, Beyer P (2008) Biofortified crops to alleviate micronutrient malnutrition.
Curr Opin Plant Biol 11(2):166–170. doi:http://dx.doi.org/10.1016/j.pbi.2008.01.007
McConnell KP, Portman OW (1952) Toxicity of dimethyl selenide in the rat and mouse. Proc Soc
Exp Biol Med 79:230–231
Meena OP, Maurya BR, Meena VS (2013a) Influence of K-solubilizing bacteria on release of
potassium from waste mica. Agric Sust Dev 1:53–56
Meena VS, Maurya BR, Bohra JS, Verma R, Meena MD (2013b) Effect of concentrate manure
and nutrient levels on enzymatic activities and microbial population under submerged rice in
alluvium soil of Varanasi. Crop Res 45(1,2 & 3):6–12
Meena VS, Maurya BR, Verma R, Meena RS, Jatav GK, Meena SK, Meena SK (2013c) Soil
microbial population and selected enzyme activities as influenced by concentrate manure and
inorganic fertilizer in alluvium soil of Varanasi. Bioscan 8(3):931–935
Meena VS, Maurya BR, Bahadur I (2014a) Potassium solubilization by bacterial strain in waste
mica. Bang J Bot 43:235–237
Meena VS, Maurya BR, Verma JP (2014b) Does a rhizospheric microorganism enhance K+ avail-
ability in agricultural soils? Microbiol Res 169:337–347
Meena RS, Meena VS, Meena SK, Verma JP (2015a) The needs of healthy soils for a healthy
world. J Clean Prod 102:560–561
Meena RS, Meena VS, Meena SK, Verma JP (2015b) Towards the plant stress mitigate the agricul-
tural productivity: a book review. J Clean Prod 102:552–553
352 D.H. Daly et al.
Meena VS, Maurya BR, Verma JP, Aeron A, Kumar A, Kim K, Bajpai VK (2015c) Potassium
solubilizing rhizobacteria (KSR): isolation, identification, and K-release dynamics from waste
mica. Ecol Eng 81:340–347
Meena VS, Meena SK, Verma JP, Meena RS, Ghosh BN (2015d) The needs of nutrient use efficiency
for sustainable agriculture. J Clean Prod 102:562–563. doi:10.1016/j.jclepro.2015.04.044
Meena VS, Maurya BR, Meena RS (2015e) Residual impact of wellgrow formulation and NPK on
growth and yield of wheat (Triticum aestivum L.) Bangladesh J Bot 44(1):143–146
Meena VS, Verma JP, Meena SK (2015f) Towards the current scenario of nutrient use efficiency in
crop species. J Clean Prod 102:556–557. doi:10.1016/j.jclepro.2015.04.030
Meena RK, Singh RK, Singh NP, Meena SK, Meena VS (2016a) Isolation of low temperature sur-
viving plant growth-promoting rhizobacteria (PGPR) from pea (Pisum sativum L.) and docu-
mentation of their plant growth promoting traits. Biocatal Agric Biotechnol 4:806–811
Meena VS, Bahadur I, Maurya BR, Kumar A, Meena RK, Meena SK, Verma JP (2016b) Potassium-
solubilizing microorganism in evergreen agriculture: an overview. In: Meena VS, Maurya BR,
Verma JP, Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture.
Springer, India, pp 1–20. doi:10.1007/978-81-322-2776-2_1
Meena VS, Meena SK, Bisht JK, Pattanayak A (2016c) Conservation agricultural practices in
sustainable food production. J Clean Prod 137:690–691
Meena RS, Bohra JS, Singh SP, Meena VS, Verma JP, Verma SK, Sihag SK (2016d) Towards the
prime response of manure to enhance nutrient use efficiency and soil sustainability a current
need: a book review. J Clean Prod 112(1):1258–1260
Meena SK, Rakshit A, Meena VS (2016e) Effect of seed bio-priming and N doses under varied
soil type on nitrogen use efficiency (NUE) of wheat (Triticum aestivum L.) under greenhouse
conditions. Biocatal Agric Biotechnol 6:68–75
Meng L, Zhang A, Wang F, Han X, Wang D, Li S (2015) Arbuscular mycorrhizal fungi and rhizo-
bium facilitate nitrogen uptake and transfer in soybean/maize intercropping system. Front Plant
Sci 6:339. doi:10.3389/fpls.2015.00339
Meplan C, Hesketh J (2012) The influence of selenium and selenoprotein gene variants on colorec-
tal cancer risk. Mutagenesis 27:177–186
Nakandalage N, Nicolas M, Norton RM, Hirotsu N, Milham PJ, Seneweera S (2016) Improving
rice zinc biofortification success rates through genetic and crop management approaches in a
changing environment. Front Plant Sci 7:764. doi:10.3389/fpls.2016.00764
Nuberger A, Okebe J, Yahav D, Paul M (2016) Oral iron supplements for children in malaria-endemic
areas. Cochrane Database Syst Rev. (2) 1–129. doi:10.1002/14651858.CD006589.pub4
Parewa HP, Yadav J, Rakshit A, Meena VS, Karthikeyan N (2014) Plant growth promoting rhizo-
bacteria enhance growth and nutrient uptake of crops. Agric Sustain Dev 2(2):101–116
Pellegrino E, Bedini S (2014) Enhancing ecosystem services in sustainable agriculture:
Biofertilization and biofortification of chickpea (Cicer arietinum L.) by arbuscular mycorrhi-
zal fungi. Soil Biol Biochem 68:429–439. doi:http://dx.doi.org/10.1016/j.soilbio.2013.09.030
Prakash S, Verma JP (2016) Global perspective of potash for fertilizer production. In: Meena VS,
Maurya BR, Verma JP, Meena RS (eds) Potassium solubilizing microorganisms for sustainable
agriculture. Springer, India, pp 327–331. doi:10.1007/978-81-322-2776-2_23
Prasad AS (2008) Zinc in human health: effect of zinc on immune cells. Mol Med 14(353):357
Prasanna R, Joshi M, Rana A, Shivay YS, Nain L (2012) Influence of co-inoculation of bacteria-
cyanobacteria on crop yield and C-N sequestration in soil under rice crop. World J Microbiol
Biotechnol 28:1223–1235
Priyadharsini P, Muthukumar T (2016) Interactions between arbuscular mycorrhizal fungi and
potassium-solubilizing microorganisms on agricultural productivity. In: Meena VS, Maurya
BR, Verma JP, Meena RS (eds) Potassium solubilizing microorganisms for sustainable agricul-
ture. Springer, India, pp 111–125. doi:10.1007/978-81-322-2776-2_8
Qin H, Brookes PC, Xu J (2016) Arbuscular mycorrhizal fungal hyphae alter soil bacterial commu-
nity and enhance polychlorinated biphenyls dissipation. Front Microbiol 7:939. doi:10.3389/
fmicb.2016.00939
16 The Role of Soil Microbes in Crop Biofortification 353
Raghavendra MP, Nayaka NC, Nuthan BR (2016) Role of rhizosphere microflora in potas-
sium solubilization. In: Meena VS, Maurya BR, Verma JP, Meena RS (eds) Potassium
solubilizing microorganisms for sustainable agriculture. Springer, India, pp 43–59.
doi:10.1007/978-81-322-2776-2_4
Rana A, Joshi M, Prasanna R, Shivay YS, Nain L (2012) Biofortification of wheat through inocula-
tion of plant growth promoting rhizobacteria and cyanobacteria. Eur J Soil Biol 50:118–126
Rana A, Kabi SR, Verma S, Adak A, Madan P, Shivay YS, Prasanna R and Nain L (2015)
Prospecting plant growth promoting bacteria and cyanobacteria as options for enrichment of
macro- and micronutrients in grains in rice-wheat cropping sequence. Cogent Food Agric 1:
1037379. doi: http://dx.doi.org/10.1080/23311932.2015.1037379
Rawat J, Sanwal P, Saxena J (2016) Potassium and its role in sustainable agriculture. In: Meena
VS, Maurya BR, Verma JP, Meena RS (eds) Potassium solubilizing microorganisms for sus-
tainable agriculture. Springer, India, pp 235–253. doi:10.1007/978-81-322-2776-2_17
Saha M, Maurya BR, Meena VS, Bahadur I, Kumar A (2016a) Identification and characterization
of potassium solubilizing bacteria (KSB) from Indo-Gangetic Plains of India. Biocatal Agric
Biotechnol 7:202–209
Saha M, Sarkar S, Sarkar B, Sharma BK, Bhattacharjee S, Tribedi P (2016b) Microbial sid-
erophores and their potential applications: a review. Environ Sci Pollut Res 23:3984–3999.
doi:10.1007/s11356-015-4294-0
Saha M, Maurya BR, Bahadur I, Kumar A, Meena VS (2016c) Can potassium-solubilising bac-
teria mitigate the potassium problems in India? In: Meena VS, Maurya BR, Verma JP, Meena
RS (eds) Potassium solubilizing microorganisms for sustainable agriculture. Springer, India,
pp 127–136. doi:10.1007/978-81-322-2776-2_9
Sathya A, Vijayabharathi R, Srinivas V, Gopalakrishnan S (2016) Plant growth-promoting actino-
bacteria on chickpea seed mineral density: an upcoming complementary tool for sustainable
biofortification strategy. 3 Biotech 6:138. doi:10.1007/s13205-016-0458-y
Sazawal S, Black RE, Ramsan M, Chwaya HM, Stoltzfus RJ, Dutta A, Dhingra U, Kabole I,
Deb S, Othman MK, Kabole FM (2006) Effects of routine prophylactic supplementation with
iron and folic acid on admission to hospital and mortality in preschool children in a high
malaria transmission setting: community-based, randomised, placebo-controlled trial. Lancet
367:133–143
Schalk IJ, Guillon L (2013) Pyoverdine biosynthesis and secretion in Pseudomonas aeru-
ginosa: implications for metal homeostasis. Environ Microbiol 15(6):1661–1673.
doi:10.1111/1462-2920.12013
Scheublin TR, Sanders IR, Keel C, van der Meer JR (2010) Characterisation of microbial commu-
nities colonising the hyphal surfaces of arbuscular mycorrhizal fungi. ISME J 4(6):752–763.
doi:10.1038/ismej.2010.5
Senés-Guerrero C, Torres-Cortés G, Pfeiffer S, Rojas M, Schüßler A (2014) Potato-associated
arbuscular mycorrhizal fungal communities in the Peruvian Andes. Mycorrhiza 24(6):405–
417. doi:10.1007/s00572-013-0549-0
Shakeel M, Afroz R, Hssan MN, Hafeez FY (2015) Root associated Bacillus sp. improves growth,
yield and zinc translocation for basmati rice (Oryza sativa) varieties. Front Microbiol 6:1286.
doi:10.3389/fmicb.2015.01286
Sharma A, Shankhdhar D, Shankhdhar SC (2016) Potassium-solubilizing microorganisms:
mechanism and their role in potassium solubilization and uptake. In: Meena VS, Maurya BR,
Verma JP, Meena RS (eds) Potassium solubilizing microorganisms for sustainable agriculture.
Springer, India, pp 203–219. doi:10.1007/978-81-322-2776-2_15
Shi Z, Mickan B, Feng G, Chen Y (2015) Arbuscular mycorrhizal fungi improved plant growth and
nutrient acquisition of desert ephemeral Plantago minuta under variable soil water conditions.
J Arid Land 7(3):414–420. doi:10.1007/s40333-014-0046-0
Shrivastava M, Srivastava PC, D’Souza SF (2016) KSM soil diversity and mineral solubilization,
in relation to crop production and molecular mechanism. In: Meena VS, Maurya BR, Verma JP,
354 D.H. Daly et al.
Wu Z, Bañuelos GS, Lin ZQ, Liu Y, Yin X, Li M (2015) Biofortification and phytoremediation of
selenium in China. Front Plant Sci 6:136. doi:10.3389/fpls.2015.00136
Yadav BK, Sidhu AS (2016) Dynamics of potassium and their bioavailability for plant nutrition.
In: Meena VS, Maurya BR, Verma JP, Meena RS (eds) Potassium solubilizing microorganisms
for sustainable agriculture. Springer, India, pp 187–201. doi:10.1007/978-81-322-2776-2_14
Yasin M, El-Medhawi AF, Jahn CE, Anwar A, Turner MFS, Faisal M, Pilon-Smits EAH (2015a)
Seleniferous soils as a source for production of selenium-enriched foods and potential of bacte-
ria to enhance plant selenium uptake. Plant Soil 386:385–394. doi:10.1007/s11104-014-2270-y
Yasin M, El-Medhawi AF, Pilon-Smits EAH, Faisal M (2015b) Selenium-fortified wheat: potential
of microbes for biofortification of selenium and other essential nutrients. Int J Phytoremediation
17:777–786. doi:10.1080/15226514.2014.987372
Yasin M, Munir I, Faisal M (2016) Can Bacillus spp. enhance K+ uptake in crop species. In: Meena
VS, Maurya BR, Verma JP, Meena RS (eds) Potassium solubilizing microorganisms for sus-
tainable agriculture. Springer, India, pp 163–170. doi:10.1007/978-81-322-2776-2_12
Yasmin S (2011) Characterization of growth promoting and bioantagonistic bacteria associated
with rhizosphere of cotton and rice. NIBGE, Faisalabad
Zahedi H (2016) Growth-promoting effect of potassium-solubilizing microorganisms on
some crop species. In: Meena VS, Maurya BR, Verma JP, Meena RS (eds) Potassium
solubilizing microorganisms for sustainable agriculture. Springer, India, pp 31–42.
doi:10.1007/978-81-322-2776-2_3
Zhang H, Sun Y, Xie X, Kim MS, Dowd SE, Paré PW (2009) A soil bacterium regulates
plant acquisition of iron via deficiency-inducible mechanisms. Plant J 58:568–577.
doi:10.1111/j.1365-313X.2009.03803.x
Zhang Y, Song Q, Yan J, Tang J, Zhao R, Zhang Y, He Z, Zou C, Ortiz-Moasterio I (2010) Mineral
element concentrations in grains of Chinese wheat cultivars. Euphytica 174:303–313
Zhang Z-Z, Lou Y-G, Deng D-J, Rahman MM, Wu Q-S (2015) Effects of common mycorrhizal
network on plant carbohydrates and soil properties in trifoliate orange–white clover associa-
tion. PLoS One 10(11):e0142371. doi:10.1371/journal.pone.0142371