Topic: Soil micro organisms, classification and their roles,

organic matter-decomposition, C:N ratio, Mineralisation and

immobilization process and humus

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Soil Biology : Deals with organisms and their function within the soil

Soil organic population: producers, decomposer and consumers

• Producers (autotrophs): synthesis their own food for growth and development
• Consumers (heterotrophs): herbivores feed on plants and carnivores feed on both
plants and animals
• Decomposers: they break down dead plants and animal residues and return vital
nutrients to soil
Classification of organism in soil
Organisms present in soil are mainly classified in to Soil flora and Soil fauna

Soil Organisms

Animals(Fauna) Plants (Flora)

Micro Flora
Micro Fauna Macro Fauna
Bacteria Macro Flora
Protozoa Earthworms
Actinomycetes Macro algae
Nematodes Ants
Fungi Plants roots
Rotifers Termites
Algae
 Soil microflora

Bacteria Fungi Actinomycetes Algae

• Blue green Algae

• Grass Green algae
Moulds yeast mushroom • Yellow green algae
• Golden brown algae (Diatoms)

• Cellulose decomposers
• Protein decomposers
• Lignin decomposers
• Humus formers

Heterotrophic Autotrophic
• Symbiotic N fixers • Nitrite formers (Nitrosomonas)
• Non-symbiotic N fixers • Nitrate formers (Nitrobacter)
• Ammonifiers • Denitrifiers
• Cellulose Decomposers • Sulphur Oxidiser
• Hydrogen Oxydiser
 Whittakkers 5 Kingdom Classification

• Monera :(prokaryotes) – Bacteria, Actinomycetes and cyanobacteria

• Protista : (single celled eukaryotes) – Protozoa and micro algae

• Fungi : Yeast, mushroom (Basediomycetes) and molds

(Deuteromycetes)

• Plantae

• Animalia
 Classification of organisms based on requirement of
molecular oxygen
Obligate aerobes: Organisms that need O2 for respiration and cannot survive without
it

e.g.: Nitrosomonas and Nitrobacter

Facultative anaerobes: Organisms which are aerobic but also adapted to grow under
anaerobic conditions using oxidised substances like NO3-, SO42- etc.

e.g.: Denitrifying bacteria (Bacillus)

Obligate anaerobes: The organisms which grow in the absence of O2

e.g.: Clostridium and Methanobacteria

 Classification based on optimum temperature for growth

Psychrophiles <10oC Arthrobactor

Mesophiles (Dominant in India) 20 -30 oC Pseudomonas, E. coli

Thermophiles (Compost pits) >45oC Thermus aquaticus

 Classification based on mode of nutrition
1. Autotrophs: Those organisms that are able to make their food using materials
from inorganic sources
• A. Chemoautotrophs: Which get energy from the oxidisation of simple
inorganic compounds (N,S or Fe) and C as CO2 (chemosynthesis) E.g.:
Thiobacillus
• B. Photoautotrophs: Organisms get energy using light and C as CO2 via the
process of photosynthesis E.g.: Algae

2. Heterotrophs: Derive energy by the oxidation of organic compounds. They cant

produce food by themselves. E.g.: Fungi
 Classification based on optimum pH for growth

Category Optimum pH for growth

Acidophiles <6.5
Neutrophiles 6.5 - 7.5
Alkaliphiles > 7.5

Classification based on food requirement

Oligotrophy: Can live in environments with very low nutrient concentration. E.g:
Lichens
Copiotropy: Organisms require higher concentration of nutrients.
E.g: E. coli
 Macroorganisms
1) Earthworms
Class: Annelida
Total biomass: 110-1100 kg/ha (furrow slice)
• The excreta deposited as cast on the surface of soil are rich in bacteria and available nutrients

• Facilitate aeration and drainage to soil

• More common in fine textured than in coarse textured soil
• Earthworm cast rich source of N, P and Ca

• Casting rate: 2600 t/ha/year

• Indigenous species used for vermi composting: Eisenia foetida, Eudrilus eugeniea, Perionyx excavates
• Place used for culturing earthworm: Wormiri

2) Termites
• Influence soil properties
• Cellulose decomposing microbes in their gut
3. Plant roots
• Rhizosphere: Term coined by Lorenz Hitlner
Unique soil environment under the influence of plant roots are rhizosphere
• Exudates: Chemical composition leaking from the roots
• Secretions: chemical compounds released through metabolism
• Mucilages: produced by the roots through bacterial degredation
• Mucigels: gelatinous layer, mixture of mucilage and oil particles
 Microorganisms
Environmental factors influencing microbial activity
1. Water
2. Temperature: most of soil organisms are mesophiles and grow well between 150C and 450C. A
temperature of 370C is considered to be optimum for most mesophiles
3. Aeration:
4. Reaction: Bacteria prefer near neutral to slightly alkaline reaction (pH 6.5 to 8.0), fungi prefer
acidic reaction (pH4.5 to 6.5) and actinomycetes prefer slightly alkaline condition.
5. Light: Direct sunlight are injurious to most of the microbes except algae
6. Food
7. Soil factors: A soil in good physical condition has good aeration and water supply capacity which
are so essential for optimum microbial activity
8. Microbial association: micro organisms interact with one another give rise to antagonistic or
symbiotic relations
 Microorganisms
1) Bacteria
Smallest and most numerous (109 per grm of soil)
Spherical - cocci
Rod shaped - bacilli
Long-spiral - spirilla
Optimum pH- 6.5-7.5
❖Largest representation in terms of species- Bacillus
❖Sulphur oxidizing bacteria- Thiobacillus
❖ Free living- Azotobacter & Azotomonas (atm. nitrogen to ammoniacal form)
❖Nitrogen fixing in acidic condition-Beijerenckia & Derxia
❖Nitrogen fixing in anaerobic condition- Clostridium
❖Nitrogen fixing through symbiosis-Rhizobium
❖Associative nitrogen fixation- Azospirillum
❖Denitrifying bacteria: Bacillus, Pseudomonas & Micrococcus
 2) Actinomycetes
▪ “Ray fungi”
▪ Characteristics of both bacteria and fungi (fungi like bacteria)
▪ More common in dry soils and undisturbed pastures and grasslands
▪ Heterotrophic
▪ Prefers- Neutral to slightly alkaline soils
▪ Organisms: Streptomyces, Micro-monospora, Nocardia and thermo-actinomyces
▪ Can withstand high temperatures
▪ The smell of a fresh soil wetted upon first rain- Production of geosmin
▪ Organic residues first added to soil are first attached by bacteria and fungi and later on by
actinomycetes (because of slow growth rate of actinomycetes)a
3) Fungi

❖Filaments- hyphae; network of hyphae- Mycelium

❖Hyphae divided by cross walls- Septa; Those without septa- coenocytic

❖Heterotrophs, devoid of chlorophyll

❖Prefers- Acidic condition

❖ Optimum soil reaction lies between pH 4.5 to 6.5. However, some soil fungi can tolerate as high as pH 9

Classes
1. Phycomycetes: Non-septate hyphae. e.g: Pythium, Rhizopus
2. Ascomycetes: Hyphae are divided by septa. e.g: Yeast
3. Deuteromycetes (Fungi imperfecti): Molds - Multiply by asexual spores, conidia. e.g: Aspergillus,
Pencillium, verticillium
4. Basidiomycetes : Mushroom
Mycorrhizal association (symbiotic association of fungi with roots of higher plants)

Endotrophic
Ectotrophic Vescicles
Hartig net Mycelium develops intracellularly
Mycelium develops intercellularly VAM (Vescicular arbuscular mycorrhizae)
Boletus, Amanita or AM
Glomus, Endogene

BENEFITS
• Increased absorption of available
nutrients
• Increased P availability
• Increased nutrient mobility
4) Algae
• Chlorophyll- containing organism, autotrophic
• Unicellular, filamentous or form colonies from single cells

Classification based on colour:

1. Cynophyta - blue green (Cyanobacteria)
Fix- 20 kg N per ha
Anabaena, Nostoc
Heterocysts: specialized cellular structure for N fixation
Anabaena azollae- Azolla -water fern

2. Chlorophyta- grass green

3. Xanthophyta- yellow green algae
4. Bacilliriophyta- golden brown algae / diatoms
5) Protozoa
• Single celled organism belong to animal kingdom
• Cyst: adverse condition- vegetative cell covers it self with a thick coating
• Flagella: long whip-like structure
• Cilia: short hair like appendages
• Pseudopodia: internal protoplasmic movement forming flexible temporary organs
6) Nematodes
• Also called thread worms
• Not much significant in organic matter decomposition
• Controlled by: Neem, karanj and sawdust

7) Viruses
• Ultramicroscopic organisms, cause many diseases
• Bacteriophages: Viruses parasiting bacteria, affect nodulation in legumes
 BIOLOGICAL INTERACTIONS OCCURING IN SOIL ECOSYSTEM
TYPE OF INTERACTION SPECIES SPECIES Nature Examples
A B
Positive interactions
Neutralism 0 0 Organism behave independently

Commensalism + 0 Partner derive benefit and the associate without Cellulose decomposers
affecting it

Protocooperation + + Both partners are mutually benefited but not Synergism b/w VAM-
obligatory Legume

Symbiosis + + Both partners are mutually benefited Rhizobium- legume

Negative interactions
Competition _ _ One species suppressed as the two species Fast growing plants V/S
struggle for the limited supply of nutrients slow growing

Ammensalism 0 _ Partners suppress the growth of other partner by Nitrobacter affected by large
producing toxins like NH3, HCN amount of NH3 that
produced after
decomposition

Parasitism + _ Association where one partner lives in or on the Bacteriophages

body of other (host)

Predation + _ Direct feeding of one organism on other Protozoa

 Organic Matter
• Organic matter is the vast array of carbon compounds in soil. Originally created by plants, microbes,
and other organisms, these compounds play a variety of roles in nutrient, water, and biological
cycles

• Organic matter can be divided into two major categories: stabilized organic matter which is highly
decomposed and stable, and the active fraction which is being actively used and transformed by
living plants, animals, and microbes.

• The main source of SOM is plant tissues.

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 Organic Matter and Decomposition

Decomposition - Definition

• Breakdown of dead plant and animal material and release of inorganic

nutrients
• Decomposition is a biological breakdown and biochemical transformation of
complex organic molecules of dead material into simpler organic and
inorganic molecules

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 Source of Organic Matter

• Plant remains
• Animal tissue and excretory products
• Cells of microorganisms
• However, plant is the main contribution to OM.

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 Organic Constituents of Plants

• Cellulose most abundant 15 to 60% of dry weight.

• Hemicellulose, 10 to 30 %
• Lignin, 5 to 30%
• Water soluble fraction include simple sugar, amino acids, aliphatic acids, 5
to 30 % of tissue weight
• Ether and alcohol-soluble constituents; fats, oils, waxes, resins and a
number of pigments.
• Proteins

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When organic tissue is added to soil, 3 general reactions takes place:

1. The bulk of the material undergoes enzymatic oxidation with carbon dioxide, energy and heat as the
major products.
2. The essential elements such as N, P and S are released and /or immobilised by a series of specific
reactions relatively unique for each element.
3. Compounds very resistant to microbial action are formed either through modification of compounds in
the original plant tissue of by microbial synthesis (Collectively these resistant compounds comprise soil
humus)
SOM→ Humic substances, non-humic substances
1.HUMIC SUBSTANCES

The humic substances makeup 60-80 % of SOM. Humic substances are characterised by aromatic, ring-type structures that
include polyphenols and polyquinones.

On the basis of solubility humic substances are divided into 3 classes:

• Fulvic acid- Lowest molecular weight among the components, both acid and alkali soluble

• Humic acid- Medium molecular weight. Alkali soluble and acid insoluble

• Humin- High molecular weight. Both acid and alkali insoluble except under the most drastic conditions.

• Fulvic acid is most susceptible to microbial attack and humin is most resistant. Depending on the environment, it may take
15-50 years to destroy fulvic acid-type compounds in the soil, hundreds of years are required to destroy humic acid.

2. NON-HUMIC SUBSTANCES

• Include all those classes of compounds occurring in plants and microorganisms that appear to have relatively definite
characteristics e.g. carbohydrates, proteins, fats, waxes, resins, pigments and low molecular weight compounds.

• Most of these could be relatively easily attacked by soil microorganisms and has a rapid turnover in the soil.
 HUMUS

• Humus is a complex and rather resistant mixture of brown or dark brown

amorphous and colloidal substances modified from the original tissues or
synthesized by the various soil organisms. Fulvic acid, humic acid and humin all
come under humus.
 Nature and characteristics of humus
• Tiny colloidal humus particles (micelles) are composed of C, H and O.
• Surface area of humus colloids are very high, generally exceeds that of silicate clays.
• Negatively charged, the sources of charge being carboxylic (-COOH) or phenolic (-
C6H5OH) groups. The extent of the negative charge is pH dependant (i.e. high at high
pH).
• Water holding capacity : 4-5 times that of silicate clays
• At high pH, CEC: 150-300 c mol/kg soil
• Low plasticity and cohesion, thus favourable effect on aggregate formation and stability.
• Black colour
• Cation exchange reactions with humus are qualitatively similar to those occurring with
silicate clays.
 Who are the Decomposers?

A.SOIL FAUNA
• earthworms, arthropods
• Fragmentation (commination) increases surface area.
• Distributes OM within soil profile
B.SOIL MICROORGANISM
• Heterotrophic bacteria, fungi
• Derive energy, carbon and nutrients from dead OM in the process they release CO2
through respiration.
RESPONSIBLE FOR BULK OF DECOMPOSITION!

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 Decomposition Process

THREE MAIN PROCESSES:

1)ASSIMILATION
• Conversion of substrates materials into protoplasmic materials
• E.g. OM carbon to microbial carbon
• E.g. protein to microbial protein
2)IMMOBILIZATION
• Conversion of inorganic form into organic.
• E.g. inorganic nitrogen from the soil converted into microbial protein.

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Cont…

3)MINERALIZATION
• Conversion of organic substance to inorganic form.
• E.g. protein from OM will be converted to inorganic nitrogen in the
soil.

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 Enzymes involved in decomposition

Organic Constituents Enzymes Microbes Involved in

Decomposition
Cellulose Bacillus,Cellulomonas
cellulase
Aspergillus,Alternaria
Hemicellulose pectinase Arthrobacter,
Pseudomonas, Bacillus
Lignin esterases, phenolases, Coriolus versicolour,
laccases, peroxidases Pleurotus ostreatus

Protein Peptidase, proteinase

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 Factors Affecting Rate Of Decomposition

Temperature • Microbial activity responds exponentially to

increased temperature until enzymes denature

• Microbial activity has optimum moisture

Moisture • Low moisture = dessication, slow diffusion

• High moisture = low O2 availability; no lignin
degradation

• Most microbes exhibit optimum activity near pH 7.

pH • Fungi most active in acid soil and bacteria in
moderate soil pH.

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 What is the C:N Ratio?

• The carbon-to-nitrogen (C:N) ratio is often used as an indication of

mineralization or immobilization whether will occur.
• The C:N ratio is the total concentration of C divided by the total concentration
of N.

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 The Changes Observed in C:N Ratio, Immobilization and
Mineralization of Organic Matter Decomposition are Depicted Below:

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 C:N ratio
Ideal soil 10-12:1
Cultivated soil 8-15:1
Humus 10:1
Micro-organism 4-9:1
Legumes 20-30:1
Manure 20-30:1
Straw 80-100:1
Sawdust 400-600:1

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 C:N Ratio Range

When surface of soil layer have a:

• C:N > 30:1 >> Immobilization highly likely to occur
• C:N < 20:1 >> Mineralization is likely to occur
• C:N between 20-30:1 >> both processes may occur but will generally in
balance

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 Substrate Quality:carbon

• Different carbon compounds are decomposed at different rates.

▪ Cellulose - faster
▪ Lignin - slower decomposition as compared to cellulose
• C:N of the OM determine the rate:
▪ High - slower, this is due to insufficient of N for microorganisms to assimilate carbon
▪ Low - faster, nitrogen is sufficient for rapid assimilation of carbon.

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 Carbon cycle
Steps of Carbon cycle:
1. CO2 used by plants for photosynthesis
2. Consumption of animals
3. Respiration by Plants, Animals and Microbes
4. Decay and Decomposition

A – Autotrophic Microorganisms
B – Respiration, Animals
C – Photosynthesis
D – Respiration, plant
E – Respiration, Microbial
End products of decomposition
• Aerobic conditions : CO2
• Anaerobic conditions : CH4
C:N ratio
The ratio of mass of carbon to the mass of nitrogen in a substances
➢Mainly controls decomposition rate in soil
➢The wide C:N ratio leads to nutrient immobilization
➢In narrow C:N ratio, plant residues decompose quickly and release
nitrates readily, mineralization
Nitrogen cycle
 Nitrogen cycle
Soil nitrogen: 95-98 % organic form
Mineralization:
Conversion of organic form to its mineral form
Ammonification: organic nitrogenous compounds are enzymatically hydrolyzed to
NH4+
Immobilization:
Conversion of inorganic to organic form
Amination: NH4+ is incorporated into amino acids
C/N ratio: < 20: 1 shows mineralization
> 30: 1 shows immobilization
Nitrification:
Conversion of ammoniacal nitrogen to nitrates
2 stage process,
i. NH4+ to NO2- (nitrite) by Nitrosomonas Optimum temperature: 30- 35°C
ii. NO2- to NO3- (nitrate) by Nitrobacter Optimum pH: 6.5-7.5
Denitrification (Dissimilatory nitrate reduction):
• Reduction of NO3- to N2 or N2O
• Pseudomonas, Bacillus and Paracoccus
Assimilatory nitrate reduction
• NO3- to NH4+, and this incorporated into proteins through amino acids-
• Enzymes involved are: Nitrate reductase, nitrite reductase, nitric oxide reductase
and nitrous oxide reductase
• Rice soils are more prone
• Organisms: Facultative anaerobes (Bacillus, Flavobacterium, Pseudomonas)
Ammonia volatilization : Loss of NH3 under alkaline condition since it is unstable
• Nitrogen fixation
Physical
Chemical
Biological
Nitrogen fixing microbes
Biological Nitrogen fixation (BNF): reducing N2 to NH3 –( enzyme –nitrogenase)
Contribution: 140 million tonnes N/ year
3 main groups
1) Symbiotic- Rhizobia (legume), Frankia (non legumes), Anabaena
2) Associative- Azospirillum, Acetobacter, Herbaspirillum
3) Free living- Azotobacter, Derxia, Cyanobacteria, Rhodospirillum, Beijerinckia
1) Legume-Rhizobium symbiosis
Tiny out growth called legume root nodule
Sesbania rostrata- nodules on the stem
Alfalfa (Medicago sativa)- 100-300 kg N/ha/year
Clover (Trifolium spp.)- 100-150 kg N/ha/year
 Common bio strains of Rhizobium
Rhizobium app. Legume inoculated

R. meliloti Medicago,Trigonella , Melilotus

R. trifoli Trifolium (Berseem)

R. Leguminosarum Pea, Lentil

R phaseoli Phaseolus

R japonicum Soybean, Ground nut, sunhemp

2) Non-legume- Frankia symbiosis
Actinomycetes- Frankia (filamentous gram positive)
Important actinorhizal plants: Alnus, Myrica, Casuarina etc.

3) Azolla-Anabaena symbiosis
Anabaena azollae - azollae with algal symbiont
Fix- 30-40 kg N/ha
Suitable temperature- 25-30°C

4) Non-specific associative N2 fixers

Azospirillum, Acetobacter, Herbasprillum- rhizosphere
Azospirillum lipoferum: Associated with roots of C4 plants like maize
Azospirillum brasilense: Associated with C3 plants like rice and wheat
Beijerinckia- leaf surface of rice, wheat, etc.

5) Free-living nitrogen fixers

Fix- 10-15 kg N/ha
Function as- Plant growth promoting rhizobacteria (PGPR)
Phosphorus cycle
 Phosphorus cycle
Mineralization: Soil phosphorus: 30-50 % organic form
Favoured by thermophilic temperature
Neutral to alkaline pH
Enzymes: Phosphatase and phytase
Organic forms: Phytin (major source), nucleic acid (Smallest source), phospholipids
C:P ratio < 200:1
Immobilization:
Inorganic forms: Iron phosphates, Calcium phosphates, Aluminium phosphates, etc.
C:P ratio >300:1
P- solubilization:
Organisms: Bacillus megatherium, Pseudomonas sp.
Produce organic acids like citric acid, glutamic acid etc., reduce pH and increase P
availability
P- mobilization:
Organism: AM fungi
 Sulphur cycle
Mineralization:
Decomposition of organic S to inorganic compounds

Assimilation/ Immobilization:
Sulphur is incorporated as S containing amino acids

Oxidation:
Reduced inorganic S compound (H2S, FeS2) is readily oxidized in soils

Organisms involved are:

• Thiobacillus denitrificans (Obligate chemolithotroph)
• T. Thioxidans
• T. Ferroxidans
• Arthrobacter
• Bacillus
• Psedomonas sp.

Reduction:
Responsible for H2S toxicity in rice (akiochi)
Organisms: Desulfovibrio desulfuricans, Desulfovibrio gigas, Desulfovibrio vulgaris, Desulfomataculum
 Other mineral elements
Iron
Fe2+ to Fe3+ under acidic condition: T. ferroxidans, Leptothrix, Gallionella
Siderophores producing microbes: Pseudomonas
Anaerobic condition: Fe 2+ electron acceptor- Bacillus, Fusarium and Alternaria
VAM also increases uptake

Manganese
Mn2+ to Mn4+: Leptothrix, Arthrobacter
 Soil enzymes
The word enzyme was coined by W. Kuhne
En means (in) and Zyme means(yeast)
Enzymes are organic catalysts produced by living organisms. Hence they are called biocatalysts.
An enzyme may be a simple protein or a complex protein.

Classification
Constitutive: it produce all the time without any substrates e.g: Urease, Dehydrogenase
Induced: formed only in the presence of a susceptible substrate e.g: Cellulase
Types of enzymes in soil
1) Oxidoreductase : oxidize a substrates by removing a pair of elements and accompanying H atom
e.g: Dehydrogenase
2) Transferases: involves in group transfer reactions
e.g: alpha-keto gluterate- aminotransferase
3) Hydrolases: involves hydrolysis reaction
e.g: Urease
Role of enzymes in nutrient cycling
C-cycle- Amylase, cellulase, lipase, glucosidases and invertase
N-cycle- Proteases, amidases, urease and deaminases
P-cycle- Phosphatases
S-cycle- Arylsulphatases
 Biofertilizers

Biologically active products containing live microorganisms on a suitable

carrier used to inoculate seed or soil
Types:
Rhizobium: inoculating legumes
PGPR: inoculating all crops
Commercially produced:
1) Rhizobium
2) Azospirillum- mainly for cereal crops
3) Azotobacter- non-symbiotic N-fixers, used for wide range of crops
4) BGA- rice
5) PSB- Pseudomonas, Bacillus, Pencillium, Aspergillus
6) VAM
• The organism which most abundant in the soil among the following
A. Bacteria
B. Fungi
C. Algae
D. Actinomycetes

Ans: option A
• Association of fungi and algae
A. Lichen
B. Parasite
C. Mycorrhiza
D. Mycoparasite

Ans: option A
• Best temperature for psychrophiles
A. < 100C
B. 20-35 0C
C. >450C
D. None

Ans: option A
• Fungi prefer condition to grow
A. Near neutral to slightly alkaline
B. Slightly alkaline
C. Acidic
D. none

Ans: option C
• Free living bacteria fix atmospheric N
A. Azotobacter
B. Acetobacter
C. Frankia
D. All

Ans: option A
• Nitrogen fixing power of legumes is considerably enhanced by application of
A. Phosphates
B. Sulphates
C. Both a & b
D. none

Ans: option A
• Heterotrophic, anaerobic and free living bacteria is
A. Pseudomonas
B. Bacillus
C. Rhizobium
D. Clostridium

Ans: option D
• Conversion of amino acid and amines into ammonia is called
A. Aminization
B. Ammonification
C. Nitrification
D. none

Ans: option B
• C:N ratio for mineralization process
A. > 30: 1
B. < 20: 1
C. > 30: 1
D. All

Ans: option B
• Optimum temperature for nitrification process
A. 30- 40 0C
B. 30- 35 0C
C. 20- 30 0C
D. 30- 50 0C

Ans: option B
• Denitrification done by which of the following bacteria
A. Pseudomonas
B. Bacillus
C. Paracoccus
D. All

Ans: option D
• The C:N ratio is the most commonly used in soils because N is the most limiting
elements.
• A microbe with a C:N ratio of 8:1 would require OM with a C:N ratio of 24:1
• The C:N ratio in lower in microorganisms = 8
• Since microbes incorporate only about 1/3 of the C metabolized into biomass, the
substrate material must have C:N ratio = 24 to satisfy the N requirement of
microbes.
• If the C:N ratio = 24, available soil N is consumed by microbes and plant available
N decrease

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