Genet Resour Crop Evol (2021) 68:1269–1282

https://doi.org/10.1007/s10722-021-01111-w(0123456789().,-volV)
(0123456789().,-volV)

REVIEW

A model for integrated approach to germplasm conservation

of Asian lotus (Nelumbo nucifera Gaertn.)
Ravi Gowthami . Neelam Sharma . Ruchira Pandey . Anuradha Agrawal

Received: 18 July 2020 / Accepted: 2 January 2021 / Published online: 23 January 2021
© The Author(s), under exclusive licence to Springer Nature B.V. part of Springer Nature 2021

Abstract Asian lotus (Nelumbo nucifera Gaertn.) is habitat loss because of global environmental change,
one of the most aesthetic ornamental aquatic indige- vanishing of wetlands, over-exploitation from its
nous species, endowed with unique biological and natural habitat, and lack of concerted management
nutritional traits. The seeds are icon of eternity due to efforts. Possible conservation strategies are proposed
remarkable longevity, known to survive in nature up with a view to draw the attention of policy makers
to 1300 years. Leaves fend off dirt and water by a and stakeholders for prioritizing this species in
phenomenon of superhydrophobicity and flowers various biodiversity-related programs of the country,
generate heat by thermoregulation to enhance polli- besides promoting it as a potential food and
nation success. Hence, lotus is an extremely nutraceutical species.
important model system to investigate the process
of aging in plants and develop designer seeds with Keywords Conservation strategies ·
high vigour, viability and longevity. The whole plant Ex situ · In situ · Indian scenario ·
is nutritionally rich with great medicinal value. While Sacred lotus
scientific attention on various aspects ranging from
basic botany to genome sequencing in lotus is
increasing worldwide, India has somewhat lagged Introduction
behind in taking due scientific, agronomic and
conservation initiatives. At present, the lotus is facing A large number of aquatic plants and their products
have been used as sources of major or minor food,
medicine and other uses in traditional societies in
R. Gowthami · N. Sharma · R. Pandey · A. Agrawal (&) India. These not only provide starch, protein, miner-
Tissue Culture and Cryopreservation Unit, ICAR-National als and vitamins, but also support livelihood of a
Bureau of Plant Genetic Resources (NBPGR), Pusa
considerable section of society living around them
Campus, New Delhi 110012, India
e-mail: anuradha.agrawal@icar.gov.in (Jain et al. 2004). The climatic and physiographic
diversity of Indian subcontinent has endowed the
R. Gowthami
e-mail: gowthami.r@icar.gov.in region with many kinds of natural water bodies
(rivers, lakes, coastal lagoons, coral reefs) and
N. Sharma
e-mail: neelam.sharma1@icar.gov.in; wetlands (peat swamps, peat lands, mangroves,
sharma.neel.59@gmail.com riverine swamps, seasonal marshes) besides innumer-
R. Pandey able human-made and intensively managed wetlands
e-mail: ruchira.pandey@icar.gov.in (aquaculture ponds, farm ponds, irrigated fields,

123
1270 Genet Resour Crop Evol (2021) 68:1269–1282

sacred groves, salt pans, reservoirs, gravel pits, Willd.], commonly named as American lotus or water
sewage farms and canals) (Gopal 2013; Bassi et al. chinquapin, is distributed in eastern and southern
2014). Unfortunately, there is little recognition of North America, Central America and West Indies.
wetland landscapes for their current and potential However, within these two species and among their
value in food, nutritional and livelihood security, crosses, [2000 cultivars/horticultural hybrids are
except paddy cultivation. A few important macro- documented, particularly in Chinese and Japanese
phytic food plants of freshwater bodies include lotus books (Tian et al. 2014). Existence of [ 400 vernac-
(Nelumbo Adans spp.), water chestnut (Trapa L. ular names prevalent in ~29 languages just for
spp.), fox nut (Euryale ferox Salisb.), wild rice Nelumbo nucifera is indicative of its importance in
(Zizania aquatica L.), water spinach (Ipomoea aquat- many societies, especially in Asia (Lim 2016). The
ica Forssk.), swamp taro (Cyrtosperma chamissonis Asian or sacred lotus is so named because of its
Merr.), Chinese water chestnut (Eleocharis dulcis venerated use in religions such as Hinduism and
(Burm.f.) Trin. ex Hensch.), water cress (Nasturtium Buddhism, prevalent in India and South-East Asian
officinale R.Br.) and water mimosa (Neptunia oler- countries (Guo 2009). It is designated by several
aceae Lour.) (Mandal and Mukhopadhyay 2010). Sanskrit terms like abja, aravinda, kamal, kamala,
While they are locally known and utilized, most nalina, padma, pankaja, puṣkara, puṇḍarīka etc.
species remain neglected in terms of diversity Though carrying specific connotation, the names
assessment, cultivation packages, breeding and relate to the unique properties of the plant and its
value-addition. There is a global concern about symbolism to life and philosophy. The immaculate
under-utilized and potential crops, and many of these lotus emergent from deep waters is correlated with
aquatic species are known only to endemic or local the idea of purity and truth in humans, which arises
tribal communities. Overharvesting, habitat alteration from good conduct and knowledge, symbolic of an
and climate change are among major drivers of enlightened mind.
decline in commercially important wild plant Research on various facets of the lotus plant like
resources used for food and medicine, posing a threat botany, taxonomy, cultivation, genetic diversity
to the species as well as livelihoods of collectors analysis, breeding of new varieties, medicinal value
(from poorest social groups) in rural area, and rarely and pharmacology, physiology of flowering, pollu-
attracts the attention of policy, makers, agriculturists tion management, metabolomics, proteomics and
or biodiversity managers (Mandal and Bar 2013). even genome sequencing has been undertaken
Such aquatic macrophytes are candidates for more world-wide (Lin et al. 2019). Lotus is an ideal
intensive research and development (R&D). Here we reference plant for study due to its unique properties
present a case study of lotus. such as superhydrophobicity and self-cleaning nature
The lotus (species of Nelumbo Adans) are rooted of lotus leaves (Wang et al. 2009), extraordinary
and emergent aquatic angiosperms with large, attrac- longevity of seeds (Shen-Miller et al. 1995; Shen-
tive flowers resembling water lily (Nymphaea L. Miller 2002), thermoregulation of lotus flowers (Li
spp.), but not related to it (Fig. 1a–c). Lotus grows and Huang 2009) and pollution management (Mishra
naturally or under cultivation in shallow (0.5–3 m) et al. 2009) (Fig. 2). In this paper we elucidate a few
freshwater lakes, ponds, tanks etc., from sea level to unique properties of the Asian lotus and bring to
1500 m elevation in the tropics and subtropics (Lim focus the paucity of efforts made towards genetic
2016). Botanically belonging to the monogeneric resource collection, evaluation and conservation in
family Nelumbonaceae, only two similar species with the Indian context.
a disjunctive distribution are commonly recognized
(Williamson and Schneider 1993). The first is N. Model plant due to unique properties (Fig. 2)
nucifera Gaertn. (syn. Nymphaea nelumbo L., N.
speciosa Willd., Nelumbium speciosum Willd.) com- Nelumbo nucifera is a rooted aquatic herb with
monly known as Asian lotus, sacred lotus or bean of horizontal creeping subterranean thick, spongy and
India, is found throughout Asia and northern Aus- densely ramified aerenchymatous rhizomes. Leaves
tralia. The second species is N. lutea Willd. [syn. N. are large and round (25–110 cm in dia), wavy, peltate
pentapetala (Walter) Fernald and Nelumbium luteum (petiole is attached to the centre of the leaf), floating

123
Genet Resour Crop Evol (2021) 68:1269–1282 1271

Fig. 1 Habit of Asian lotus (Nelumbo nucifera). a Natural population of plants in a pond in Shillong, Meghalya, India; b A pink-
white lotus single whorled variety in full bloom; c Fruit receptacle with individual fruits at different maturity stages

or raised above the water (up to 200 cm), on terete, slightly longer than gynoecium. The individual pistils
glabrous or prickly, hollow, long petioles (1–3.5 m). are embedded in a yellow flattened carpellary
Flowers are solitary, hermaphrodite, large, showy, turbinate receptacle (5–10 cm in dia). Shaped like a
(10–42 cm in dia), sweet-scented and borne by spout of a watering can, the fleshy receptacle forms
lengthy scape (flower stalk) reaching well above the composite structures that hold the individual fruits
leaves, glabrous or parsely spinulate. Tepals (20 or (15–40), botanically termed etaerio of achenes (dry
more) are caducous, pink or white, oblong elliptic to fruit which does not open on its own and contains a
obovate and spirally arranged. Stamens are numerous single seed). The shower-head-like receptacle that
(100–200) with slender filaments and linear anthers, houses the fruits is yellow during flower opening,

123
1272 Genet Resour Crop Evol (2021) 68:1269–1282

Fig. 2 Unique properties of Asian lotus

green during fruit development, and dark brown at lutea to a subspecies (N. nucifera subsp. lutea). In
full maturity (Fig. 1c). Each fruit is oblong to ovoid poetry, art and philosophy literature, lotus plants are
with thick pericarp, hardened (bony), green when often interchangeably used as water lilies (Nymphaea
immature and brown to black at maturity. Seed has a spp., family Nymphaeales). However, the two are
reddish brown testa, two yellowish–white mealy clearly distinguishable by their leaves, flowers and
cotyledons and a stalked green embryo (Goel et al. fruits.
2001; Lim 2016). Amongst flowering plant species, Asian lotus
The American lotus (N. lutea) is distinguished seeds have exceptional longevity and period of
from Asian lotus (N. nucifera) in having yellow rather viability, with documented life span ranging from
than white or pink petals, and outermost petals 100 to 1300 years (Shen-Miller et al. 1995; Shen-
persisting longer at the base of the developing fruit. Miller 2002). Fruits are adapted to alternating wet-
The American plant also has wider fruits (length/ dry conditions, and to prolonged periods of quies-
width ratio\1.25) compared to Asian Lotus (length/ cence followed by germination and growth. Unique
width ratio[1.5) (Small and Catling 2005). Aside morho-physiology and underlying genetic disposition
from morphological differences, anther development render the plant with this feature. The dry lotus fruit
has also been recorded to be different between the pericarp, which surrounds the seed, is hard and
two species (Zhang et al. 2019a). Some taxonomists impervious to water, and this water- and air-tight
recognize only one species of Nelumbo, reducing N. architecture is a significant factor contributing to its

123
Genet Resour Crop Evol (2021) 68:1269–1282 1273

longevity (Shen-Miller et al. 1995). Histologically, mechanisms that controls thermal metabolism during
the pericarp is composed of an outermost epidermis, pollination is important from functional biology
two thick layers of palisade cells with a small acid- perspective.
resistant fibrous zone sandwiched between them, and The water repellency property of lotus, also known
an inner parenchymatous cell layer (Shen-Miller as the “lotus effect” has attracted the attention of
2002). A high level of enzyme superoxide dismutase plant scientists as well as material scientists over the
(SOD) activity in seed has been shown to extend seed years (Marmur 2004; Ensikat et al. 2011). The
longevity, and germination rate is influenced by adaxial leaf surfaces have convex papillae coated
levels of oxidized/reduced forms of glutathione and with nanoscopic agglomerated wax crystals tubules
ascorbic acid (Li et al. 2000). Asian lotus seeds can (10–100 nm). The morphology of the papillae greatly
also withstand extremely high temperatures due to reduces the contact area to water drops. The chemical
the presence of several thermostable proteins like composition of the epicuticular wax with high
annexin, and repair enzymes, such as L-isoaspartyl content of nonacosanediols leads to the growth of a
methyltransferase (Chu et al. 2012; Shen-Miller et al. dense layer of very small wax tubules with a
2013). These proteins help in the protection and permanently hydrophobic surface. This characteristic
repair of cellular damage through processes like of ultra-hydrophobicity ensures that the leaf upper
antioxidation, anaerobic glycolysis, microbial epidermis is not covered by water, thus maintaining
defense, membrane maintenance, stress repair and the normal function of its stomata (Ensikat et al.
tolerance. Three genes (NnMT2a, NnMT2b and 2011). Two wax biosynthesis-related genes (NnCER2
NnMT3) that encode metallothioneins, (cysteine-rich and NnCER2-LIKE) cloned from lotus and trans-
small proteins involved in ROS scavenging) have formed in Arabidopsis thaliana (L.) Heynh., resulted
been found to be highly expressed in germinating in an alteration of the cuticle wax structure in
lotus seeds and are significantly upregulated in inflorescence stems, and proved their function in the
response to high salinity and oxidative stresses (Zhou biosynthesis of the extra-long fatty acids (Yang et al.
et al. 2012). Thus, lotus is an extremely important 2018). More studies on the lotus leaf chemical
model system to investigate the process of aging and compositions and structure might be very helpful in
senescence in plants and to develop ‘designer seeds’ producing materials with super hydrophobicity and
with high vigour, viability and longevity. self-cleaning features (Lin et al. 2019).
Among other intriguing properties, the Asian lotus
is one among the few plants on the earth which has
the ability to generate heat and regulate its own Multifarious uses of Asian lotus
temperature like birds and mammals (Li and Huang
2009). During anthesis, flowers can adjust the tem- Lotus is a multipurpose plant, well known for its use
perature by heat production, retaining a as vegetable, medicinal and ornamental. All the parts
comparatively favorable temperature between 30 of plant (rhizome, nodes, roots, seed, young shoots
and 36 °C, when ambient temperature oscillates and leaves, petioles, petal, stamen, and pericarp) are
between 10 and 45 °C (Seymour et al. 1998). The consumed raw or cooked as vegetables throughout
flowers take in more oxygen and release more carbon Asia. Petals are used as garnish or brewed along with
dioxide as the night air cools, converting more stamens and leaves to make herbal-tea; flower-stalks
carbohydrates to energy, just like a shuddering are stir-fried or used in soups and in salads. The
animal to stay warm. There is evidence that ther- rhizomes are used in soups, stir-fried, chopped and
moregulation is ecologically important and might be fried (similar to potato chips) or turned into lotus-
a reward to insect pollinators that would benefit from starch. Seeds can be eaten raw, roasted or ground and
a high body temperature while remaining in the made into flour. Seeds consist mainly of carbohy-
flower (Seymour et al. 1998; Li and Huang 2009). It drates and proteins, with low fat content, and
has been demonstrated that the receptacle, stamens nutritional compounds such as polyphenols and
and petals produce heat independently, and the source amino acids (Zhang et al. 2015). The leaves are used
of heating is due to a cyanide-resistant alternative as plates to wrap and steam food such as rice. The
oxidase pathway (Grant et al. 2010). Study of dried seed-pods are used in flower arrangements and

123
1274 Genet Resour Crop Evol (2021) 68:1269–1282

the whole plant is invaluable to the cosmetics dysentery and also used to improve appetite, memory
industry. and immunity (Chen et al. 2019). The lotus stem is
All parts of lotus have been used for medicinal used for the treatment of vomiting, diarrhea, leprosy,
purposes in various systems of medicine including strangury, skin diseases, and nervous exhaustion
folk medicines, Ayurveda, Chinese traditional medi- (Sridhar and Bhat 2007). Roots of the plant have
cine, and oriental medicine (Fig. 3). The whole plant been used to cure piles, dysentery and also for skin
is used as astringent, emollient, diuretic and sudorific infections and ringworms (Chopra et al. 1956). The
and cardiotonic (Sridhar and Bhat 2007). Lotus lotus flower is used for treatment of fever, diarrhea,
leaves are used to treat summer heat syndrome in premature ejaculation, hyperdipsia and bloody dis-
China and Japan, and to treat obesity in China (Ono charges (Mukherjee et al. 2009). Lotus seeds are used
et al. 2006). Rhizome is used for the treatment of to treat tissue inflammation, including insomnia,
diarrhea, chronic dyspepsia, chronic liver cirrhosis, palpitations, cancer, fever, poor digestion, skin

Fig. 3 Medicinal properties of various parts of plants of Asian lotus

123
Genet Resour Crop Evol (2021) 68:1269–1282 1275

diseases, leprosy, diarrhea, enteritis (Chopra et al. Genetic diversity and genomics
1956; Nadkarni 1982). A Chinese drug called Lian Zi
Xin prepared by using lotus seed embryos is helpful Aquatic plant species are characterized by the ability
in insomnia, nervous disorders, cardiovascular dis- to reproduce both sexually and asexually, which can
eases and high fevers (Chen et al. 2007). Seed powder influence the genetic diversity of populations growing
and honey is used to treat cough (Mukherjee et al. in different types of environments, as is also the case
2009). Seeds after peeling, coring, grinding, boiled of lotus (Nomura et al. 2015). Based on their distinct
into a syrup or paste, is used as an ingredient of attributes and commercial use and for the conve-
traditional pastries and desserts in China, Japan, nience of production/breeding, lotus cultivars are
Korea and India (Moro et al. 2013). Lotus honey, classified into three major groups named as flower,
considered like ‘Amrita’ in Ayurveda to cure many seed and rhizome lotus (Guo et al. 2007). Seed and
ailments, is also used to treat eye infections (Mukher- rhizome lotus are produced for their edible tissues,
jee et al. 2009, Mandal and Brar 2013). whereas, flower lotus is popular for its flower color
In India, most of the research related to lotus is and shape (Li et al. 2018). It is generally accepted
focused on its pharmacological studies (Sridhar and that Nelumbonaceae consists of two disjunct species,
Bhat 2007). Pharmacological activities of different N. nucifera and N. lutea with same chromosome
parts of lotus are ascribed for human health viz., number (2n=16) and there is no interspecific repro-
analgesic, antibacterial, anticancer, antidiabetic, ductive barrier. Geographical isolation and easy
antidiarrheal, antidermatophytic, antifertility, antifun- hybridization between species has caused many
gal, antiinflammatory, antimicrobial, antiobesity, complications in terms of species circumscription,
antioxidant, antiparkinsonian, antiplatelet, antipyre- genetic variation assessment and status of extant
tic, antithrombotic, aphrodisiac, antisteroidogenic, germplasm. Extensive studies on genetic diversity
antistress, diuretic, hepatoprotective, hypoglycemic, assessment of lotus germplasm have been carried out
immunomodulatory, psychopharmacological activi- by several researchers from China, Japan and Thai-
ties and nutritional value (Indrayan et al. 2005; land using morphological, cytological, biochemical
Paudel and Panth 2015). and molecular markers (see Lin et al. 2019 and
In addition to its nutraceutical, medicinal and references therein). Results revealed that rich genetic
pharmacological importance, N. nucifera also has variation present in lotus genepool has been greatly
phytoremediation properties for heavy metal pollu- influenced by adaptation of the plants to different
tion in aquatic bodies. It has been successfully environments due to geographical separation as well
demonstrated to remove up to 96% of Cu, upon as artificial selection (Chen et al. 2008; Hu et al.
exposure to Cd and Cu pollution in water bodies 2012; Yang et al. 2012). What is clear from the
(Mishra et al. 2009). In a recent study, the plant has studies so far is that a combination of markers and
shown very promising potential for treating run-off holistic studies are required to assess the genepool of
fertilizer in the natural environment, as it could lotus, which displays abundant phenotypes in nature
significantly reduce the biological oxygen demand as well as under cultivation.
(BOD) and chemical oxygen demand (COD) of The draft genomes of two lotus genotypes ‘China
contaminated surface water (Rasid et al. 2019). Liu Antique’ and ‘Chinese Tai-zi’ were sequenced,
et al. (2012) isolated a gene [phytochelatin synthase1 assembled and released in 2013 (Ming et al. 2013;
(NnPCS1)] coding for an enzyme involved in the Wang et al. 2013). It revealed that out of all the plants
synthesis of phytochelatins, which in turn plays sequenced so far, sacred lotus bears the closest
important role in heavy metal detoxification. Ara- resemblance to the ancestor of all eudicots (Wang
bidopsis transgenic plants expressing NnPCS1 gene et al. 2013). Chloroplast genome sequencing by
accumulate more Cd than the wild type. Hence, lotus maximum likelihood (ML) trees of 79 combined
is recommended as one of the aquatic plants for chloroplast protein-coding genes of 133 taxa from 56
growing in the ponds, to help in discharging the orders revealed that Nelumbo is a member of basal
industrial effluents and purify water in a natural eudicots, sister to Platanus. The inverted copies (IR)
manner. structural variations i.e., IR expansions/contractions
within the basal eudicots occurred independently or

123
1276 Genet Resour Crop Evol (2021) 68:1269–1282

were caused by independent insertions and deletions pinkish-white and pink. Pinkish-white flowers are
(Wu et al. 2014). A phylogenetic tree built using the more in demand compared to the other two varieties
sequences of 41 mitochondrial protein-coding genes (Mandal and Bar 2013). White flowers are used for
also revealed N. nucifera to be a sister to all other the preparation of garlands. The pink variety is not
eudicots and its mitochondrial genome having usually cultivated by farmers due to low yield. In all
retained ancient gene content and gene clusters, has the three varieties, there are two types of flowers,
high levels of RNA editing and low levels of namely double flowers (100–150 petals per flower)
chloroplast-derived fragment insertions (Gui et al. and single flowers (25–50 petals per flower) (Mandal
2016). Subsequently comprehensive transcriptomic and Bar 2013).
analysis revealed 36,385 protein-coding genes in
‘Chinese Tai-zi’ (Gui et al. 2018) and 32,121 protein-
coding genes in ‘China Antique’ (Zhang et al. Research and development in India
2019b). Analysis of the lotus genome has fast-tracked
identification of genes controlling important traits In view of the significance of lotus plant, an ad-hoc
such rhizome yield and quality, seed size and project on exploration, collection and cataloguing of
nutritional profile, flower morphology, and flowering sacred lotus and studies on its reproductive biology
time for crop improvement (Ming et al. 2013). and seed technology, was carried out under the All
Recently, three functional markers (FMHXK-E1, India Coordinated Research Project (Medicinal and
FMGBSS-I8 and FMAGPL-I1) have been identified Aromatic Plants, MAP) at College of Agriculture,
for lotus rhizome with suitable starch content for Vellanikkara. A total of 49 accessions from different
molecular assisted selection in the early stage to districts of Kerala and Kanyakumari district of Tamil
shorten the breeding life and improve breeding Nadu (Krishikosh 2019). Subsequently, under the All
efficiency (Cheng et al. 2020). Bioinformatics sup- India Network Research Project on MAP, 42 acces-
port in the form of a database named as LOTUS-DB sions of lotus from different areas of Kerala and
(http://lotus-db.wbgcas.cn) has also been generated Tamil Nadu were assembled by the ICAR-National
for comprehensive information on annotation, gene Research Centre for MAP, Boriavi. These exhibited
function and expression (Wang et al. 2015). These wide variability in flower colour, petal number and
developments are expected to accelerate the utiliza carpels (AICRP 2008). In recent years, ICAR-Indian
tion of genetic resources, QTL analysis, map-based Institute of Vegetable Research (IIVR), Varanasi,
gene cloning, and molecular breeding with MAS in also initiated collection and conservation of lotus
lotus, besides facilitating proteomic and transcrip (ICAR-IIVR 2018). In all the above cases, germ-
tomics studies on lotus. plasm was conserved ex situ in cement tanks or
plastic tubs and diversity recorded at morphological
level only.
Cultivation of lotus in India The CSIR-National Botanical Research Institute
(NBRI), Lucknow, initiated a project on collection,
So far, the lotus flowers, rhizomes, leaves available in introduction, acclimatization, documentation and
the Indian markets are collected predominantly from multiplication of racial variants of N. nucifera under
natural habitats, resulting in low regeneration of the a Botanic Garden Flagship Program (Goel et al.
species. The income from lotus is high, as it has 2001). Reportedly a national collection of 60 species/
multiple uses, including having a sacred value in races of Nelumbo were augmented. These included 35
religious ceremonies. However, the species’ exis- indigenous species/races collected from 11 States/
tence is under threat due to the conversion of its Union territories of India and 25 exotic races
growing areas into paddy fields, construction of ring- introduced from Australia, Brazil, Germany, Thai-
bands for fishing, siltation from the surrounding land, U.K. and U.S.A. However, currently only 15
uplands due to overland flow, development projects species/cvs/races of N. nucifera (lotus) and Nym-
and urbanization (Jain et al. 2004). phaea spp. (water lilies) are listed as being available
Usually, three varieties of lotus flowers are found (NBRI 2020). NBRI developed a package of prac-
in India, based on petal colour, such as white, tices for growing lotus successfully in cement

123
Genet Resour Crop Evol (2021) 68:1269–1282 1277

concrete tanks by having a clay soil stratum up to important to preserve in situ many populations to
45 cm thick at the bottom of the tank and maintaining conserve genetic diversity and allow evolutionary
the water level from 11.5 m. Under a recent Niche- processes. However, ex situ conservation is also
Creating Flagship Program, CSIR-NBRI proposes to much required because of the low genetic diversity
identify and select germplasm with high nutraceutical within populations. Seeds and rhizomes can be used
and medicinal properties for commercial exploitation for ex situ conservation.
in India, undertake analysis of genetic diversity of So far, all the attempts of ex situ conservation have
Asian lotus, develop agro-technologies/models for been made by establishment of field genebanks in
propagation and cultivation of identified elite germ- cement tanks or plastic tubs. But maintenance of lotus
plasm in different agro-climatic conditions of India in such conditions is labour-intensive, prone to high
through multi-location trials and development of a risk from pests, diseases, scarcity of water and other
few nutraceutical products using different plant parts environmental vagaries. Though the lotus seeds are
of lotus, targeting the global market (NBRI 2020). recognized to have exceptional longevity and viabil-
ity among extant flowering plant species, they have
not been conserved in seed genebanks. Lotus seeds
Germplasm conservation of lotus in India: a trans- are known to have long period of dormancy (Shen-
situ approach Miller et al. 1995; Presannakumari et al. 2000; Shen-
Miller 2002). Inspite of extensive studies conducted
Conventional germplasm conservation programs nor- on seed longevity in lotus, there is paucity on data
mally view in situ and ex situ methods as two related to seed storage behavior (including germina-
complimentary approaches, although in practice tion as per genebank standards) for the purpose of
either one is adopted, depending on the biology of germplasm conservation. Such studies would deter-
the species. In the face of unprecedented climatic mine the optimal storage regime, and seeds can
change and disasters, social conflict, and political possibly be conserved either at ambient (~25 °C), low
uncertainty, the concept of ‘trans situ’ conservation to (~ − 4 to − 20 °C) or ultra-low (− 180 to − 196 °C)
safeguard genetic diversity and accessibility for crop temperatures, in seed genbanks and cryobanks.
improvement is being widely promulgated (Riordan However, there have not been any systematic studies
and Nabhan 2019). Trans situ conservation dynam- on the seed biology, seed conservation, or seed
ically integrates initiatives to combine multiple in situ cryopreservation, leading to a base collection for
and ex situ measures, from conservation to research posterity. Another option for conservation of the
to education, spanning local to global scales. Ex situ rhizome lotus would be use of plant tissue culture
collections (representing genetic and environmental techniques using vegetative explants (Agrawal et al.
diversity) serve as an important insurance policy for 2019). Tissue culture protocols in several aquatic
increasingly threatened in situ populations. These angiosperms have been developed in India (Mohan
“backups” require maintenance, and themselves are Ram and Agrawal 1997). Micropropagation in lotus
vulnerable to genetic erosion through genetic drift has been reported using mature and young embryos,
and viability selection. Ex situ samples and in situ rhizome buds, shoot apical meristems, immature
populations are never exact analogs of one another plumule by direct multiplication or through callus
and diverge genetically through time (Riordan and (Arunyanart and Chaitrayagun 2005; Shou et al.
Nabhan 2019). 2008; Mahmad et al. 2014; Liu et al. 2016). Effect of
Based on the results of genetic diversity in lotus various plant growth regulators, explant size, season
germplasm in East and South-East Asia it is evident of explant collection, temperature and photoperiod on
that populations of wild lotus, cultivars, hybrids and in vitro lotus shoot, callus and somatic embryos have
species constitute unique genetic combinations and been estimated. Being an aquatic plant, a liquid–solid
need attention from conservation point of view. double-layer medium that simulates the natural
In situ conservation is usually the preferred strategy habitat of lotus has been shown to be beneficial
for most wild plant species because it allows natural (Liu et al. 2019), similar to micropropagation in
populations to continue to be exposed to evolutionary water chestnut (Agrawal and Mohan Ram 1995).
processes (Bellon et al. 2017). Accordingly, it is Detailed studies are required to optimize in vitro

123
1278 Genet Resour Crop Evol (2021) 68:1269–1282

Fig. 4 Schematic representation of the proposed model of trans-situ conservation for Asian lotus

123
Genet Resour Crop Evol (2021) 68:1269–1282 1279

conservation protocols in lotus, to aid its germplasm using ecological niche modeling and metapopulation
conservation. modelling needs to be undertaken to understand the
Needless to reiterate that trans-situ conservation present status of distribution and spatial analyses in
strategy would entail a priori information of genetic patterns of species richness. Thereafter, systematic
diversity within and among the populations. Studies studies on genetic diversity, conservation biology
undertaken in South-East and East Asia have clearly (including seed storage behavior, in vitro conserva-
shown that among the wild populations of N. tion and cryopreservation) leading to scientific
nucifera, there is high genetic diversity, which collection and a trans-situ complimentary conserva-
indicates that outcrossing is predominant. Between tion program within India (Fig. 4) require funding
the clonally propagated collections, diversity is also priority and thrust. Simultaneously, there is need for
high. Hence, collecting and conservation of only a development of package of practices or standard
few populations may not adequately protect the agro-techniques for cultivation in submerged and
genetic variation within the species. Therefore, amphibious conditions (with minimum water require-
several populations throughout the entire range of ment) as the natural populations face high extraction
the species in the country should be considered for pressures. Rotational harvesting based on scientific
conservation. In the context of in situ conservation studies on regeneration capacity and yield aspects can
and restoration genetics this finding is of critical lead to development of proper harvesting regimes.
importance. In situ conservation of populations which Breeding efforts to develop specific cultivars for
have high genetic diversity and high levels of particular use (flower, rhizome or seeds), short
outcrossing would be particularly advisable. duration varieties, identification or development of
Developing and applying suitable ex situ conser- cut flower type with increased shelf life also need
vation protocol (at least two or more) using whole attention from professional breeders and
plants, seeds, embryos/embryonic axes, in vitro horticulturists.
explants, pollen and DNA would depend on protocols Management of Nelumbo nucifera species in the
developed, infrastructure and expertise. Collaboration wild habitats requires greater awareness. Conserva-
between institutes having germplasm and expertise tion education among users and collectors also need
would be very beneficial. The complementarity, to be given due attention. Bioprospecting of lotus
redundancy and synergy gained through a trans-situ seeds as an alternate protein supplement and potential
model would facilitate in the conservation and pharmaceutical source is promising from nutritional
availability of genetic diversity for plant breeding and health security perspective. Although its
and crop research. Figure 4 provides a schematic nutraceutical value is established, further precise
representation of the proposed model of trans-situ exploration of value-added compounds might be
conservation for Asian lotus. beneficial in health promotion. As lotus seeds have
potential nutraceutical advantage, blending its flour
with other nutritionally rich legumes (e.g. soybean) or
Suggested future focus for Research millets (e.g. finger millet) will be of immense value to
and Development (R&D) in Asian lotus develop low-cost proteinaceous and health food
supplements to combat malnutrition as well as
Wetlands per se, and plant species that they harbor in specific ailments. Till date several uses of lotus plant
particular, perform valuable functions but are often have been elucidated in terms of both food and
ignored. The Asian lotus is one such example. medicine, and with increased fundamental studies
Despite its immense potential, the plant has failed using ‘omics’ technologies, formulation of new
to receive attention of agriculturists, food and nutri- products and usages can be expected to emerge in
tion industry and continues to be extracted from the future.
natural populations. Attention of scientists, breeders,
entrepreneurs and policy makers is drawn on aspects
of Asian lotus R&D that require concerted efforts. In Acknowledgements Sincere thanks are accorded to Director,
ICAR-NBPGR, New Delhi, for providing necessary support.
the first instance, population inventory, habitat We gratefully acknowledge photographs courtesy to Late Prof.
distribution mapping, species distribution mapping

123
1280 Genet Resour Crop Evol (2021) 68:1269–1282

H.Y. Mohan Ram, University of Delhi; Miss Magudeeshwari Crit Rev Food Sci 59:sup1. https://doi.org/10.1080/
P., PhD Scholar, CAU, Meghalaya; Miss Lepakashi N.M., 10408398.2018.1553846
Agriculture Officer, Karnataka; and Mr. Subhash S., Scientist, Cheng T, Zheng X, Xie K, Liu J, Zheng X, Jin S, Diao Y, Hu
ICAR-CPRI, Shimla. Z, Wang J (2020) Development and identification of three
functional markers associated with starch content in lotus
Authors’ contributions AA and GR conceived the idea, (Nelumbo nucifera). Sci Rep 10:1–7. https://doi.org/10.
collated information and wrote the draft. NS and RP provided 1038/s41598-020-60736-6
constructive feedback and editorial inputs. All authors have Chopra RN, Nayar SL, Chopra IC (1956) Glossary of Indian
reviewed and approved the manuscript. medicinal plants. Council of Scientific Industrial
Research, New Delhi
Compliance with ethical standards Chu P, Chen H, Zhou Y, Li Y, Ding Y, Jiang L, Tsang EW, Wu
K, Huang S (2012) Proteomic and functional analyses of
Conflict of interest The authors declare that they have no Nelumbo nucifera annexins involved in seed thermotol-
conflict of interest. erance and germination vigor. Planta 235:1271–1288.
https://doi.org/10.1007/s00425-011-1573-y
Ensikat HJ, Ditsche-Kuru P, Neinhuis C, Barthlott W (2011)
Superhydrophobicity in perfection: the outstanding prop-
References erties of the lotus leaf. Beilstein J Nanotechnol 2:152–
161. https://doi.org/10.3762/bjnano.2.19
Agrawal A, Mohan Ram HY (1995) In vitro germination and Goel A, Sharma SC, Sharga AN (2001) The conservation of the
micropropagation of water chestnut (Trapa sp.). Aquatic diversity of Nelumbo (lotus) at the National Botanical
Bot 51:135–146. https://doi.org/10.1016/0304-3770(95) Research Institute, Lucknow, India. Bot Gard Conserv
00471-B News 3:52–54
Agrawal A, Singh S, Malhotra EV, Meena DPS, Tyagi RK Gopal B (2013) Future of wetlands in tropical and subtropical
(2019) In vitro conservation and cryopreservation of Asia, especially in the face of climate change. Aquatic Sci
clonally propagated horticultural species. In: Rajasekha- 75:39–61. https://doi.org/10.1007/s00027-011-0247-y
ran PE, Ramanatha Rao V (eds) Conservation and Grant N, Miller R, Watling J, Robinson S (2010) Distribution
utilization of horticultural genetic resources. Springer, of thermogenic activity in floral tissues of Nelumbo
Singapore, pp 529–578. https://doi.org/10.1007/978-981- nucifera. Funct Plant Biol 37:1085–1095. https://doi.org/
13-3669-0 10.1071/FP10024
AICRP (2008) Indian Council of Agricultural Research, Gui S, Peng J, Wang X, Wu Z, Cao R, Salse J, Zhang H, Zhu Z,
Directorate of Information and Publications of Agricul- Xia Q, Quan Z, Shu L (2018) Improving Nelumbo nuci-
ture. Research Achievements of AICRPs on Horticulture, fera genome assemblies using high-resolution genetic
2002–2007. Krishi Anusandhan Bhavan, New Delhi, p 51 maps and BioNano genome mapping reveals ancient
Arunyanart S, Chaitrayagun M (2005) Induction of somatic chromosome rearrangements. Plant J 94:721–734.
embryogenesis in lotus (Nelumbo nucifera Geartn.). Sci https://doi.org/10.1111/tpj.13894
Hortic 105:411–420 Gui S, Wu Z, Zhang H, Zheng Y, Zhu Z, Liang D, Ding Y
Bassi N, Kumar MD, Sharma A, Pardha-Saradhi P (2014) (2016) The mitochondrial genome map of Nelumbo
Status of wetlands in India: a review of extent, ecosystem nucifera reveals ancient evolutionary features. Sci Rep
benefits, threats and management strategies. J Hydrol Reg 6:30158. https://doi.org/10.1038/srep30158
Stud 2:1–19. https://doi.org/10.1016/j.ejrh.2014.07.001 Guo HB (2009) Cultivation of lotus (Nelumbo nucifera Gaertn.
Bellon MR, Dulloo E, Sardos J, Thormann I, Burdon JJ (2017) ssp. nucifera) and its utilization in China. Genet Resour
In situ conservation—harnessing natural and human- Crop Evol 56:323–330. https://doi.org/10.1007/s10722-
derived evolutionary forces to ensure future crop adapta- 008-9366-2
tion. Evolut Appl 10:965–977. https://doi.org/10.1111/ Guo HB, Li SM, Peng J, Ke WD (2007) Genetic diversity of
eva.12521 Nelumbo accessions revealed by RAPD. Genet Resour
Chen Y, Fan GR, Wu HL, Wu YT, Mitchell A (2007) Sepa- Crop Evol 54:741–748. https://doi.org/10.1007/s10722-
ration, identification and rapid determination of liensine, 006-0025-1
isoliensinine and neferine from embryo of the seed of Hu J, Pan L, Liu H, Wang S, Wu Z, Ke W, Ding Y (2012)
Nelumbo nucifera Gaertn. by liquid chromatography Comparative analysis of genetic diversity in sacred lotus
coupled to diode array detector and tandem mass spec- (Nelumbo nucifera Gaertn.) using AFLP and SSR mark-
trometry. J Pharm Biomed 43:99–104. https://doi.org/10. ers. Mol Biol Rep 39:3637–3647. https://doi.org/10.1007/
1016/j.jpba.2006.06.016 s11033-011-1138-y
Chen Y, Zhou R, Lin X, Wu K, Qian X, Huang S (2008) ISSR ICAR-IIVR (2018) Annual Report 2017-18. ICAR-Indian
analysis of genetic diversity in sacred lotus cultivars. Institute of Vegetable Research Institute, Varanasi, India,
Aquatic Bot 89:311–316. https://doi.org/10.1016/j.aqua pp 18–19
bot.2008.03.006 Indrayan AK, Sharma S, Durgapal D, Kumar N, Kumar M
Chen G, Zhu M, Guo M (2019) Research advances in tradi- (2005) Determination of nutritive value and analysis of
tional and modern use of Nelumbo nucifera: mineral elements for some medicinally valued plants from
phytochemicals, health promoting activities and beyond. Uttaranchal. Curr Sci 89:1252–1255

123
Genet Resour Crop Evol (2021) 68:1269–1282 1281

Jain A, Singh RS, Singh HB (2004) Economic evaluation of Mohan Ram HY, Agrawal A (1997) Tissue culture of aquatic
lotus Nelumbo nucifera Gaertn. cultivation in Sanapat plants and its application. In: Ravishankar GA, Venka-
lake, Manipur Valley. Natural Prod Radiance 3:418–421 traman LV (eds) Biotechnological applications of plant
KrishiKosh (2019) An Institutional Repository of Indian tissue and cell culture. Oxford & IBH Publishing Co.,
National Agricultural Research System. https://krishikosh. New Delhi, pp 3–13
egranth.ac.in/ . Accessed 3 Dec 2019 Moro CF, Yonekura M, Kouzuma Y, Agrawal GK, Rakwal R
Li J, Shi T, Huang L, He D, Nyong ATM, Yang P (2018) (2013) Lotus–A source of food and medicine: Current
Systematic transcriptomic analysis provides insights into status and future perspectives in context of the seed pro-
lotus (Nelumbo nucifera) seed development. Plant Growth teomics. Int J Life Sci 7:1–5
Regul 86:339–350 Mukherjee PK, Mukherjee D, Maji AK, Rai S, Heinrich M
Li JK, Huang SQ (2009) Flower thermoregulation facilitates (2009) The sacred lotus (Nelumbo nucifera) - phyto-
fertilization in Asian sacred lotus. Ann Bot 103:1159– chemical and therapeutic profile. J Pharm Pharmacol
1163. https://doi.org/10.1093/aob/mcp051 61:407–422. doi:https://doi.org/10.1211/jpp.61.04.0001
Li YD, Wei YN, Xu BM (2000) Study on the ABA content and Nadkarni AK (1982) The Indian Materia Medica. vol, 1.
SOD activity in ancient lotus and modern lotus seeds. Popular Prakashan Pvt, Bombay
Chinese Bull Bot 17:439–442 NBRI (2020) Lotus-Niche creating programme. https://nbri.res.
Lim TK (2016) Nelumbo nucifera. In: Lim TK (ed) Edible in/lotus/ . Accessed 24 Apr 2020
Medicinal and Non-Medicinal Plants, Volume-11, Modi- Nomura K, Harada T, Saotome N, Kubota M, Katori M (2015)
fied Stems, Roots, Bulbs. Springer, Switzerland, pp 55– Clonal structure of two wild lotus (Nelumbo nucifera
109. https://doi.org/10.1007/978-3-319-26062-4 Gaertn.) populations inferred from Amplified Fragment
Lin Z, Zhang C, Cao D, Damaris RN, Yang P (2019) The latest Length Polymorphism fingerprints. Plant 3:8–14
studies on lotus (Nelumbo nucifera)—an emerging horti- Ono Y, Hattori E, Fukaya Y, Imai S, Ohizumi Y (2006)
cultural model plant. Int J Mol Sci 20(15):3680. Antiobesity effect of Nelumbo nucifera leaves extract in
https://doi.org/10.3390/ijms20153680 mice and rats. J Ethnopharmacol 106:238–244
Liu Q, Zhang D, Liu F, Qin M, Tian D (2019) Micropropa- Paudel KR, Panth N (2015) Phytochemical profile and bio-
gation of Nelumbo nucifera ‘Weishan Hong’ through logical activity of Nelumbo nucifera. Evid Based
germfree mature embryos. In Vitro Cell Dev Biol Plant Complement Altern Med 2015:789124. https://doi.org/10.
55:305–312. https://doi.org/10.1007/s11627-019-09984-4 1155/2015/789124
Liu Y, Chaturvedi P, Fu J, Cai Q, Weckwerth W, Yang P Presannakumari KT, Suma VA, Girija T (2000) Investigation
(2016) Induction and quantitative proteomic analysis of on seed dormancy in sacred lotus. In: Proceedings of
cell dedifferentiation during callus formation of lotus National Seminar in plant biology, Central Plantation
(Nelumbo nucifera Gaertn. spp. baijianlian). J Proteom Crops Research Institute, Kasargod, India, p 77
131:61–70. https://doi.org/10.1016/j.jprot.2015.10.010 Rasid NA, Naim MN, Man HC, Bakar NA, Mokhtar MN
Liu Z, Gu C, Chen F, Yang D, Wu K, Chen S, Jiang J, Zhang Z (2019) Evaluation of surface water treated with lotus plant
(2012) Heterologous expression of a Nelumbo nucifera Nelumbo nucifera. J Environ Chem Engineer 7(3):103048.
phytochelatin synthase gene enhances cadmium tolerance https://doi.org/10.1016/J.JECE.2019.103048
in Arabidopsis thaliana. Appl Biochem Biotech 166:722– Riordan EC, Nabhan GP (2019) Trans situ conservation of crop
734. https://doi.org/10.1007/s12010-011-9461-2 wild relatives. Crop Sci 59:2387–2403. https://doi.org/10.
Mahmad N, Mat Taha R, Othman R, Saleh A, Hasbullah NA, 2135/cropsci2019.06.0356
Elias H (2014) Effects of NAA and BAP, double-layered Seymour R, Schultze-Motel P, Lamprecht I (1998) Heat pro-
media, and light distance on in vitro regeneration of duction by sacred lotus flowers depends on ambient
Nelumbo nucifera Gaertn. (lotus), an aquatic edible plant. temperature, not light cycle. J Exp Bot 49:1213–1217.
Sci World J 12:74518. https://doi.org/10.1155/2014/ https://doi.org/10.1093/jxb/49.324.1213
745148 Shen-Miller J (2002) Sacred lotus, the long-living fruits of
Mandal RN, Mukhopadhyay PK (2010) Little-known treasures China Antique. Seed Sci Res 12:131–143. https://doi.org/
of wetlands need conservation. Sci Repoter 47:27–29 10.1079/SSR2002112
Mandal RN, Bar R (2013) The sacred lotus - an incredible Shen-Miller J, Lindner P, Xie Y, Villa S, Wooding K, Clarke
wealth of wetlands. Resonance 18:732–737. https://doi. SG, Loo RR, Loo JA (2013) Thermal-stable proteins of
org/10.1007/s12045-013-0094-3 fruit of long-living Sacred Lotus Nelumbo nucifera Gaertn
Marmur A (2004) The Lotus effect: Superhydrophobicity and var. China Antique. Trop Plant Biol. https://doi.org/10.
metastability. Langmuir 20:3517–3519. https://doi.org/10. 1007/s12042-013-9124-2
1021/la036369u Shen-Miller J, Mudgett MB, Schopf JW, Clarke S, Berger R
Ming R, Vanburen R, Liu Y, Yang M, Han Y, Li LT, Zhang Q, (1995) Exceptional seed longevity and robust growth:
Kim MJ, Schatz MC, Campbell M (2013) Genome of the ancient sacred lotus from China. Am J Bot 82:1367–1380.
long-living sacred lotus (Nelumbo nucifera Gaertn.). https://doi.org/10.1002/j.1537-2197.1995.tb12673.x
Genome Biol 14:R41. https://doi.org/10.1186/gb-2013- Shou SY, Miao LX, Zai WS, Huang XZ, Guo DP (2008)
14-5-r41 Factors influencing shoot multiplication of lotus (Nelumbo
Mishra V, Pathak V, Tripathi B (2009) Accumulation of cad- nucifera). Biol Plant 52:529–532. https://doi.org/10.1007/
mium and copper from aqueous solutions using Indian s10535-008-0103-7
lotus (Nelumbo nucifera). AMBIO J Human Environ Small E, Catling PM (2005) Blossoming treasures of biodi-
38:110–112 versity: 15. Asian Lotus (Nelumbo nucifera) and

123
1282 Genet Resour Crop Evol (2021) 68:1269–1282

American Lotus (N. lutea), spiritual symbols, valuable lotus constructed using novel SSR markers derived from
foods, and successful invaders. Biodivers 6:25–30. the genome of sequenced cultivar. BMC Genom 13:653.
https://doi.org/10.1080/14888386.2005.9712756 https://doi.org/10.1186/1471-2164-13-653
Sridhar KR, Bhat R (2007) Lotus—a potential nutraceutical Yang X, Wang Z, Feng T, Li J, Huang L, Yang B, Zhao H,
source. J Agric Technol 3:143–155 Jenks MA, Yang P, Lü S (2018) Evolutionarily conserved
Tian DK, Mo HB, Zhang WW, Huang X, Li C, Xu YY (2014) function of the sacred lotus (Nelumbo nucifera Gaertn.)
Progress on international lotus registration and construc- CER2-LIKE family in very-long-chain fatty acid elonga-
tion of international Nelumbo database. Acta Hortic tion. Planta 248:715–727. https://doi.org/10.1007/s00425-
1035:79–85. https://doi.org/10.17660/ActaHortic.2014. 018-2934-6
1035.9 Zhang D, Chen Q, Liu Q, Liu F, Cui L, Shao W, Wu S, Xu J,
Wang J, Chen H, Sui T, Li A, Chen D (2009) Investigation on Tian D (2019a) Histological and cytological characteri-
hydrophobicity of lotus leaf: Experiment and theory. Plant zation of anther and appendage development in Asian
Sci 176:687–695. https://doi.org/10.1016/j.plantsci.2009. lotus (Nelumbo nucifera Gaertn.). Int J Mol Sci 20:1015.
02.013 https://doi.org/10.3390/ijms20051015
Wang K, Deng J, Damaris RN, Yang M, Xu L, Yang P (2015) Zhang Y, Lu X, Zeng S, Huang X, Guo Z, Zheng Y, Tian Y,
LOTUS-DB: an integrative and interactive database for Zheng B (2015) Nutritional composition, physiological
Nelumbo nucifera study. Database 2015:bav023. functions and processing of lotus (Nelumbo nucifera
https://doi.org/10.1093/database/bav023 Gaertn.) seeds: a review. Phytochem Rev 14:321–334.
Wang Y, Fan G, Liu Y, Sun F, Shi C, Liu X, Peng J, Chen W, https://doi.org/10.1007/s11101-015-9401-9
Huang X, Cheng S, Liu Y (2013) The sacred lotus genome Zhang Y, Nyong AT, Shi T, Yang P (2019b) The complexity of
provides insights into the evolution of flowering plants. alternative splicing and landscape of tissue-specific
Plant J 76:557–567. https://doi.org/10.1111/tpj.12313 expression in lotus (Nelumbo nucifera) unveiled by Illu-
Williamson PS, Schneider EL (1993) Nelumbonaceae. In: mina- and single-molecule real-time-based RNA-
Kubitzki K, Rohwer JG, Bittrich V (eds) Flowering plants sequencing. DNA Res 26:301–311. https://doi.org/10.
dicotyledons. The families and genera of vascular plants. 1093/dnares/dsz010
vol 2, Springer, Berlin, pp 470–473. https://doi.org/10. Zhou Y, Chu P, Chen H, Li Y, Liu J, Ding Y, Tsang EW, Jiang
1007/978-3-662-02899-5_55 L, Wu K, Huang S (2012) Overexpression of Nelumbo
Wu Z, Gui S, Quan Z, Pan L, Wang S, Ke W, Liang D, Ding Y nucifera metallothioneins 2a and 3 enhances seed germi-
(2014) A precise chloroplast genome of Nelumbo nucifera nation vigor in Arabidopsis. Planta 235:523–537.
(Nelumbonaceae) evaluated with Sanger, Illumina MiSeq, https://doi.org/10.1007/s00425-011-1527-4
and PacBio RS II sequencing platforms: Insight into the
plastid evolution of basal eudicots. BMC Plant Biol
Publisher’s Note Springer Nature remains neutral with
14:289. https://doi.org/10.1186/s12870-014-0289-0
regard to jurisdictional claims in published maps and
Yang M, Han Y, VanBuren R, Ming R, Xu L, Han Y, Liu Y
institutional affiliations.
(2012) Genetic linkage maps for Asian and American

123