Plant Science Today (2018) 5(2): 44-54 44

https://dx.doi.org/10.14719/pst.2018.5.2.382

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Plant Science Today

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Mini Review

Somaclonal variations for crop improvement: Selection

for disease resistant variants in vitro
Veena S Anil*, Spurti Bennur and Savitha Lobo

Department of Plant Biotechnology, Gandhi Krishi Vignana Kendra (GKVK), University of Agricultural Sciences (UAS), Bangalore
560065, India

Article history Abstract

Received: 07 February 2018
Somaclonal variations (SV) are genetic or epigenetic changes induced in plant cell and
Accepted: 04 March 2018
Published: 01 April 2018 tissue culture. Induction of somaclonal variation, is an alternate approach to
conventional breeding and transgenic approaches to introduce desirable genetic
variability in the gene pool. SVs that occur spontaneously in culture induce changes in a
range of plant characters. However, the probability of improving a key agronomic trait
© Anil et al (2018) such as disease resistance can be cumbersome when left to chance alone. The efficiency
of developing disease resistant SVs is better with the imposition of an appropriate in
vitro selection pressure. Selection agents that have been applied include pathogen
elicitors, pathogen culture filtrate and purified pathotoxins. This method of SV selection
Editor has been successful in enhancing disease resistance in several crops and it is an accepted
K K Sabu biotechnological approach with tremendous potential for crop improvement.

Keywords
Publisher
Somaclonal variations (SV); biotechnology; crop improvement; in vitro selection, disease
Horizon e-Publishing Group
resistance; culture filtrate; pathotoxins

Citation
Correspondence
Veena S Anil
Anil V S, Bennur S, Lobo S. Somaclonal variations for crop improvement: Selection
for disease resistant variants in vitro. Plant Science Today 2018;5(2):44-54.
[email protected] https://dx.doi.org/10.14719/pst.2018.5.2.382

Introduction the development of transgenic plants and the in

Biotic stresses are major constraints that contribute vitro selection of somaclonal variations, become
to yield losses, and to the failure of crops realizing viable alternatives to support the breeding
their full yield potential. Breeding is the programmes for crop improvement. Among these
conventional approach to introduce genetic changes alternatives, transgenic approach is yet to gain
for crop trait improvement including disease public acceptance, and has to adhere to stringent
resistance. Resistance may arise from the biosafety regulations of the country in which the
introduction of resistance (R) genes, which remains transgenic crop is developed, tested and cultivated.
effective over a period of time until overcome by an The somaclonal variant approach is based on
ever evolving pathogen, thus limiting the shelf life of changes resulting from internal mutations and thus
the effectiveness of the cultivars. In addition, does not face acceptability issues nor pose any
breeding approaches can fail due to lack of genetic known biosafety concerns. SVs can be considered an
variability in a crop species or its wild relatives. In alternate source of exploitable variation induced
this scenario, biotechnological approaches such as in cell and tissue culture. This mini review briefly

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Plant Science Today (2018) 5(2): 44-54 45

highlights the resistance mechanisms existing in the coexistence of different plant species in a
plants and details the approaches used in natural ecosystem. During domestication, crops
enhancing disease resistant traits in crop plants. have gained in yield potential, but have lost out on
The review focuses on somaclonal variations as a resistance traits of their wild ancestors. Crop
potential technology in crop trait improvement in disease contributes to an average loss of 26% of
general and more specifically for developing the global crop production annually (2). Crop
disease resistant variants. cultivation relies on a few in-bred disease
resistance genes and on excessive application of
pesticides to manage pathogens. Despite their
Disease Resistance
effectiveness pesticides have deleterious
Plants, are constantly exposed to various environmental consequences and the development
pathogens and pests but very few are successful in of genetically resistant cultivars becomes a
establishing an infection and causing disease. paramount goal. It is indeed a major challenge to
Occurrence of disease in nature is rare as plants attain food security, in the backdrop of population
have evolved multiple, sophisticated and explosion, climate change, soil salinity, drought
overlapping mechanisms of defense including and soil erosion. Developing long-lasting and
defensive structures, toxins, antimicrobials, broad-spectrum disease resistance in crops will
barriers such as callose, suberins, waxes, and contribute in the quest to attain yield stability and
more specific adaptive defense such as non-host food security.
defense, vertical-race-specific and horizontal
multigenic resistance.
Approaches to enhance disease resistance
i) Breeding approach: Plant breeders focus a
Types of disease resistance
significant part of their effort on selection and
i) Vertical resistance: The term was first coined development of disease resistant plant lines. Crop
by Vanderplank during 1963 (1) to describe single- varieties and wild ancestral species with inherent
gene resistance. In vertical resistance, plant disease resistance genes are generally the source
possesses single genes for resistance [example, of resistance in breeding programmes. A disease-
Resistance (R) genes], while the pathogen susceptible desirable crop cultivar is crossed with
possesses single genes for pathogenicity [example a variety with suitable resistance trait to obtain
Avirulence (Avr) genes], which interact and populations that segregate for the traits of the
recognize each other at the protein level. This is parents. Crossing includes cumbersome
known as the gene-for-gene relationship between phenotypic selection, and methods such as marker
a pathogen and host, and is the basis of vertical assisted selection, backcross breeding, pedigree
resistance. It is qualitative resistance or race and bulk methods.
specific resistance regulated by major genes which Breeding for disease resistance has been
is effective but can be easily overcome by new an ongoing process since the domestication of
races of the pathogen. In other words, the plants but it requires persistence, takes many
pathogen can mutate its Avr genes to escape years to develop and the resulting cultivar may
recognition by the host R gene, thus by-passing an revert to susceptibility in a few years. This is
effective resistance response in the host. because pathogens are under natural selection
for enhancing their pathogenicity. Thus with
ii) Horizontal resistance: This term was also time and the right mutations pathogens can
coined by Vanderplank (1) and represents a overcome the plant’s resistance. Moreover new
quantitative or durable resistance, controlled by pathogens maybe introduced to the area,
several genes. It is also sometimes referred to as changes in cultivation practice can trigger
generalized resistance. Horizontal resistance and incidence of new diseases, and sometime
horizontal pathogenicity are entirely independent breeding for other characters can disrupt the
of each other in genetic terms, that is, there is no disease resistance present in the parent
gene-for-gene relationship in this phenomenon. varieties. Many a times the crop species may
The polygenic resistance genes of horizontal have limited genetic variability making breeding
resistance provide the plants with defensive programs unviable. In some instances related
structures or toxins that slow down or stop the wild species may possess the required resistance
pathogenic infection. The resistance may not be as genes, however crossability barriers prevent the
precise as the race specific vertical resistance and use of such putative wild parents. To overcome
develops at a slower rate. However it is a durable some of these short comings and to hasten the
resistance and does not break down to new races process of developing new cultivars,
of the pathogen, as does vertical resistance. conventional breeding is now integrated to other
modern methods such as genetic engineering,
somatic hybridization, double haploid and multi-
Need for Disease Resistant Crops parent advanced generation inter-cross (MAGIC)
Agriculture involves large areas of monoculture of populations (3).
genetically identical crops, which is very unlike

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Table 1. Examples of in vitro selection for disease resistance in crop plants

No Crop Selective agent Resistance References

1 Carica papaya Sporangial Phytophthora palmivora (46)

suspension

2 Glycine max Culture filtrate Septoria glycines (47)

3 Gossypium hirsutum Culture filtrate Fusarium oxysporum, Alterania macrospora (48)

4 Hordium vulgare Fusaric acid Helminthosporium sativum (49)

5 Lycopersicon Culture filtrate Pyrenochaeta lycopersici (50)

esculentum

6 Medicago sativa Culture filtrate Fusarium oxysporum f. sp. medicaginis (51,52)

7 Oryza sativa Culture filtrate Helminthosporium oryzae (53)

8 Psidium guajava Cell free fitrate Pencillium vermosonii (54)

9 Psidium guajava Culture filtrate Fusarium oxysporum (55)

10 Triticum aestivum Deoxy-nivaenol Fusarium graminearum (56)

11 Tobacco Methionine Psuedomonas syringae (21)

suifoximine

12 Saccharum officinarum Phytotoxin Colletotrichum falcatum (57)

13 Potato Fungi filtrate Phytophthora (58)

culture

14 Ground nut Phytotoxin Carcosporidium peronatum (35)

selection

15 Pigeon Pea Culture filtrate Fusarium odum (36)

16 Potato Culture Filtrate Phytophthora infestans (22)

17 Mango Culture Filtrate Colletotrichum (59)

gloeosporioides

18 Sunflower Culture filtrate Alterneria helianthi (39)

19 Lemon Pathogen Toxin Phoma tracheiphila (60)

20 Garlic Culture filtrate Sclerotium cepivorum (61)

21 Ginger Spore suspension Fusarium oxysporum (40)

of pathogen

ii) Transgenic approach: Transgenic plants species, however as the inserted gene is a
possess genetically engineered recombinant DNA recombinant DNA they are also considered as
and are considered as genetically modified GMOs. Inserting a combination of genes (gene
organisms (GMO). This approach allows the stacking) in a plant is more beneficial and
introduction of a new trait that does not occur productive as the introduced trait may last longer.
naturally in the species due to the artificial Most genetically modified plants are generated by
insertion of a gene or genes to the genome. The using the Agrobacterium tumefaciens mediated
transgene may originate from a related plant or transformation method or by the biolistic method
from a completely different species (from within (Particle gun method) (4-5) which have both
and across Kingdoms) or may even be a proved effective.
completely synthetic gene. Cisgenic plants, on the Over the last two decades genetic
other hand, have inserted gene(s) from the same engineering and transgenic technology have been

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Fig 1. Schematic representation for a general methodology to select resistant Somaclonal variants

used in tandem to develop disease resistant crops. release of transgenic crops. These regulations are
Genetic engineering has the potential to solve essential; nevertheless they slow down the
some of the problems of conventional breeding by development of transgenic crops. Transgenic
inserting multiple genes that can confer long plants have not been accepted in several societies
lasting broad-spectrum resistance. Recent many a times due to unsubstantiated notions or
advances in molecular techniques have unravelled sentiments, such as reluctance to ingest DNA from
some of the intricacies of the multifaceted nature virus, bacteria or animal sources. Thus the
of plant resistance mechanisms, which has in turn technology has not made inroads to the larger crop
led to more sophisticated transgenic approaches to cultivation of several countries. However,
enhance resistance. A broader and in depth transgenic food crops have been used for decades
understanding of plant resistance along with in the Americas without any proven ill-effects, and
transcriptomics, proteomics, metabolomic and the advantages of the technology are many. It has
protein interaction studies have thrown up several great potential for developing superior traits for
candidate genes from plants, bacteria, viruses and yield, abiotic and biotic stress tolerance, and can
fungi that potentially can enhance resistance. contribute to the much needed world’s food
These candidate genes can be constitutively over stability.
expressed, induced to express under biotic threat,
tissue-specifically expressed, knocked out or
iii) Somaclonal Variant Approach: Tissue culture
silenced by RNAi to obtain the desired resistance
derived plants are referred to as somaclones and
trait. The technology has been successful in
tissue culture derived plants exhibiting
several crops and has potential to reduce losses
divergences are referred to as somaclonal variants
incurred by biotic stress.
(6). Callus cultures can be used to recover somatic
There have been instances of transgenics mutants because the in vitro culture milieu
failing to perform, often due to the way in which encourages the division of individual cell and
the gene is expressed. Constitutively over regeneration of whole plant. Somaclonal variants
expressed transgenes adversely affect plant size or can be somatically or genetically stable. The
seed production. Failure could also result due to genetically stable variations can be termed
disruption of an important endogenous gene by mutations. However, because of the possibility of
insertion of the transgene. In addition, there are reversible epigenetic variations this area broadly
concerns that 1) transgenic crops may cause uses the term ‘variations’ instead of ‘mutations’ (7).
allergies in people, 2) the marker antibiotic
Any change in the DNA sequence are
resistance genes, integrated in these crops can
heritable and important for crop improvement. On
induce resistance to antibiotics, leading to super
the other hand, epigenetic changes are temporary
bugs, 3) transgenic crops can cause damage to the
and reversible and not heritable (8). Genetically
environment, by affecting beneficial microbiota
stable SVs can result due to point mutations,
and insects, and 4) there is the fear that genetic
alterations in chromosome number and structure,
modifications maybe unintentionally transferred
recombinations, methylation of DNA sequences,
to other related species via the pollen. These
deletions and transpositions in nuclear,
concerns have led to the establishment of strict
mitochondrial or chloroplast genomes (9,10).
guidelines and regulation for the development and
These genomic changes may result in stable

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48 Plant Science Today (2018) 5(2): 44-54

Fig 2. Schematic representation for a tissue culture methodology to evaluate stability of somaclonal variants

alterations, which are transmitted sexually to the In India, somaclonal variation has been the
progeny. Such SV based mutations can be very biotechnological approach to produce commercial
advantageous for improving a cultivar (11). varieties. CIMAP (Central Institute for Medicinal
and Aromatic Plants), Lucknow has released ‘Bio
13’ a somaclonal variant of Citronella java, a
Somaclonal variations in crop improvement
medicinal plant. This SV, Bio-13, has 37% higher oil
Stable SVs have been generated in several plant and 39% higher citronellol as compared to control
species (12) and is a way to create variations and varieties (16). A somaclonal variant of the B.
broaden the germplasm pool. In fact, somaclonal juncea variety ‘Varuna’ has been released for
variation is a simple alternate technology aiding commercial cultivation as ‘Pusa Jai Kisan’
breeders for enhancing genetic variability (http://nrcpb.org/content/varieties-developed). The
relatively rapidly in crops that are difficult to new variety has bolder seeds and yield advantage
breed or have low genetic variability (11,13). over the parent variety Varuna. Gupta et al. (2002)
The SV breeding practice has led to several (17) developed a superior somaclonal variant of
cultivars that have been successfully released with Rose-scented Geranium at CIMAP, India. Arun and
improved traits including plant architecture, coworkers (2003) (18) generated SVs (R 2, R3 and R4
disease resistance, yield, appearance, and abiotic generations) from immature embryos of two
stress tolerance. Some examples include Yidan No. spring wheat varieties, HUW-206 and HUW-234.
6 maize (Zea mays L.) with improved grain quality, These SVs displayed improved characters such as
CIMAP/bio-13 aromatic grass (Cymbopogon resistance to spot blotch disease and enhanced
winterianus Jowitt) with enhanced oil yield, He Zu yield over the source varieties. A high sugared and
No. 8 a wheat (Triticum aestivum L.) variant with high yielding SV of sugarcane (CoC 671), Co 94012,
high yield, rice variant DAMA with resistance to released as Phule Savitri in Maharastra (19), has
Picularia spp. ‘Ono’ somaclonal variant of better sucrose content and resistance to red rot
sugarcane is resistant to eye-spot disease, Fiji and smut diseases.
disease and downy mildew, and generated from
the susceptible cultivar ‘Pindar’ (14). A sweet
Selection pressure to induce disease resistance
potato somaclonal variant cultivar ‘Scarlet’ is
in vitro
comparable to the parent cultivar in yield and
disease resistance, but shows a more desirable The conventional and more cumbersome method
darker and more stable skin colour (15). of obtaining SVs involves the field screening of
Somaclonal variants of St. Augustine grass showed resistance in a large population of plants, raised
stable desirable variations during vegetative through in vitro callus cultures. A more targeted
propagation (12). In vitro selection of desirable approach is the regeneration of disease resistant
traits that have been commercially exploited in plants by generating resistant callus cultures
horticulture varieties through somaclonal selected on fungal toxin or culture filtrates (20-22).
variations are enumerated in a review by Krishna Somaclonal variation has applications in plant
et al. in 2016 (11). breeding and genetic improvement and generation
of such novel variants can be enhanced by

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Table 2. Examples for molecular detection of Somaclonal variants in different crops

Common name Species Source of variation Detection method Reference
Tea Camellia sinensis (L.) Embryogenic culture, RFLP, RAPD, microsatellite (62)
O. Kuntze genotype markers
Lemon Citrus limon (L.) Callus culture, gamma- Chromosome count, RAPD (63)
Burm rays
Coffee Coffea arabica L. Embryogenic culture AFLP (64)
Lemmon grass Cymbopogonflexuosus Callus culture, 2,4-D, RAPD (65)
(Nees ex Steud.) number of subcultures
Will.Watson
Jamrosa Cymbopogon hybrid Callus culture, 2,4-D Morphology, RAPD (42)
Strawberry Fragaria L. 6-benzylaminopurine Morphology, RAPD (66)
Soybean Glycine max (L.) Merr. Embryogenic culture, 2,4- RAPD (67)
D
Cotton Gossypium hirsutum Callus culture, 2,4-D + Chromosome count, RAPD, (68)
L. kinetin, duration in microsatellite markers
culture
Wild barley Hordeum Callus culture Sequence-specific amplification (69)
brevisubulatum polymorphism (S-SAP), AFLP,
(Trin.) Link MSAP
Barley Hordeum vulgare L. Callus culture Inter-retrotransposon (70)
amplified polymorphism
(IRAP), microsatellite markers
Banana Musa acuminata L. Genotype, explant source, RAPD (71)
number of subcultures
Banana Musa acuminata L. Explant AFLP, MSAP (72)
Banana Musa acuminata L. Number of subcultures, RAPD, inter-retrotransposon (73)
activation of transposable amplified polymorphism
element (IRAP), susceptibility to
fusarium wilt disease
Banana Musa acuminata L.cv. Somatic embryo culture cDNA-RAPD (74)
Rasthali screened for Fusarium
wilt resistance
Rice Oryza sativa L. Callus culture, genotype, Morphology, RAPD (75)
duration in culture
Rice Oryza sativa L. Callus culture, DNA RAPD, microsatellite markers (76)
demethylation using 5-
azacytidine
Orchids Phalaenopsis Hsiang Fei Embryogenic culture cDNA-AFLP (77)
Pea Pisum sativum L. Callus culture, genotype RAPD, microsatellite markers (78)
Sugarcane Saccharum L. hybrid Callus culture Morphology, chromosome (79)
count, isozyme patterns
Rye Secale cereale L. Embryogenic culture Biochemical tests, AFLP (80)
Potato Solanum L. Genotype Microsatellite markers (81)
Potato Solanum tuberosum L. Callus culture, duration Microsatellite markers (82)
of culture
Potato Solanum tuberosum L. Embryogenic culture Chromosome count, AFLP (83)
Sorghum Sorghum bicolor L. Explant Microsatellite markers (84)
Cocoa Theobroma cacao L. Embryogenic culture Cleaved amplified polymorphic (85)
sequence (CAPS)

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providing a suitable in vitro selection pressure (Helianthus annuus L.) to Alternaria helinathi was
(23). Exposure to phytopathotoxin, pathogen-wall improved by exposure of callus cultures to
material or secreted elicitors, has been a proven Alternaria culture filtrates (39). Bhardwaj et al. in
selection pressure to obtain disease resistant 2012 (40) obtained resistant mutants of ginger
somatic variants of several crop plants (11,24). In (Zingiber officinale Rosc.) against wilt pathogen
such a selection method, the cultures are selected (Fusarium oxysporum f. sp. zingiberi Trujillo) by in
by gradually increasing concentrations of culture vitro selection approach. More, recently, Krishna
filtrate or phytotoxin or by a constant challenge et al. (2016) (11) and Dehgahi and Joniyas, in 2016
with either. Following such selection (Fig 1), the (24) have reviewed the success of somaclonal
calli can be screened further on medium variants and the use of pathogen toxin, wall
containing a higher concentration of the toxin or elicitors and culture filtrate as a selection pressure
culture filtrate. The stability of the putative to generate disease resistance in different crops.
cultures can be tested by passing the cultures
through several subcultures in recovery medium,
Molecular detection of somaclonal variants
devoid of the selection pressure. This is followed
by screening of the recovered calli on Efficient detection of alterations is essential to
phytopathotoxic medium (Fig. 2). The ability to identify somaclonal variants that might possess
survive and grow on such screening medium will useful agronomic traits. SVs have been detected
give an indication of the stability of the adaptation and analyzed using different methods, including
observed in the putative SVs. morphological, physiological, resistance
evaluation, cytological, biochemical and molecular
Somaclonal breeding programmes have
methods. At the molecular level, variations arise
been effective in developing disease-resistant
from changes in chromosome number or
crops (25,26,22,27). Phytopathotoxins have been
structure, or from subtle changes in the DNA itself
used as a proven selection pressure in callus
(41).
cultures. The scientist Carlson in 1973 (21) was the
first to report in vitro selection of callus for Molecular analysis of variations at the DNA
breeding purpose. Maize plants resistant to level is sensitive and will enable the detection of
Helminthosporium maydis were generated by changes that are not obvious at the morphological
Gengenbach et al. (1977) (28), using culture filtrate level. Moreover, molecular techniques enable
selection pressure on callus cultures. Later, detection of variants in the callus or juvenile
Behnke (1979) (29) regenerated Late Blight stages as opposed to morphological and
resistant potato plants from culture filtrate- physiological methods wherein regenerated or
exposed callus (22,29). Tomato (Lycopersicon adult plant response are measured. Most
esculentum) plants with improved resistance to importantly molecular detection methods will help
Fusarium (Fusarium oxysporum) wilt were identify genetically stable variations among a
obtained by exposing calli to fusaric acid (30). group of SVs that might have arisen due to either
Potato plants resistant to Early Blight (caused by epigenetic changes or due to subtle but stable
Alternaria solani) and Late Blight (caused by divergence at the DNA level. Molecular methods
Phytophthora infestans) were regenerated from are useful tools to analyze the degree of
protoplasts of potato (S. tuberosum L.) varieties divergence in SVs from the source calli or plant
‘Rssset Burbank’ (31) and ‘Bintje’ (32). This material.
approach has also been used in rice to select for Random Amplified Polymorphic DNA (RAPD)
resistance against brown spot disease (33). Cerato markers are the most commonly employed
and co-workers have shown that plants markers used to detect genetic variations in
regenerated from potato cells selected in vitro with somaclonal variants (42,43). In addition ISSR
culture filtrate of Phytophthora infestans exhibited markers (44), AFLP, RFLP and microsatellite DNA
improvement in resistance as compared with the markers have been used for molecular analysis of
source plants (34). variations induced in tissue culture (Table 2) (45).
In India, the selection pressure approach has For effective detection and evaluation of
been used in groundnut against Carcosporidium somaclonal variants, it is preferable to use a multi
peronatum (35), and in chickpea cell lines against thronged approach as shown in Table 2.
Fusarium oxysporum sp. cicero (36). Thakur et al. Somaclonal variants selected specifically for
(2002) (37) carried out in vitro selection and disease resistance are detected and evaluated
regeneration of carnation (Dianthus cayophyllus using a combination of biochemical methods,
L.) plants resistant to culture filtrate of Fusarium tissue culture screening, disease resistance assays,
oxysporum f. sp. dianthi. Rao et al. (2006) (36) cell viability tests and molecular marker methods.
developed Pigeon Pea cell lines and regenerated The selected putative SV calli are first screened on
plantlets from callus tolerant to culture filtrate of media containing toxic levels of the
Fusarium odum Buttler. Saxena et al. (2008) (38) phytopathotoxin or CF and selected based on the
generated leaf blight-resistant Pelargonium response of the SV compared to the source calli
graveolens (rose scented Geranium) plants by (Fig 2). Further the calli cells may be analyzed by
selection of callus with culture filtrate of vital stains such as Trypan blue to evaluate the
Alternaria alternate. The resistance of sunflower percentage viability in the calli in the screening

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plates or even after direct exposure to the References

pathogen. The calli can further be evaluated for 1. Vanderplank JE. Plant diseases: Epidemics and
the levels of defense enzyme activity such as control. Academic Press, New York and London,
superoxide dismutase, catalase, peroxidase as a 349pp.1963.
response to exposure to the toxin or the pathogen. 2. Oerke EC. Crop losses to pests. Agricultural Sci.
The shortlisted putative somaclonal variants 2006;144:31-43.
showing significant resistance response to the https://doi.org/10.1017/S0021859605005708
pathogen or its elicitor, are further analyzed using 3. Bandillo N, Raghavan C, Muyco PA, Sevilla MAL,
molecular marker methods to ascertain that LobinaI T, Ermita CJD, Tung CW, Mccouch S,
indeed the resistance adaptation observed is due Thomson M, Mauleon R, Singh RK, Gregorio G,
to genetic changes. Redoña E, Leung H. Multi-parent advanced
generation inter-cross (magic) populations in rice:
progress and potential for genetics research and
Drawbacks of the SV technology breeding. Rice. 2013; 6: 11.
Selection of disease resistance in cell cultures is a https://doi.org/10.1186/1939-8433-6-11
rather simple process. However, the regeneration 4. Estrella H, Simpson J, Trujillo M. Transgenic plants:
of plantlets from such somaclonal variant calli is an historical perspective. Methods Mol Biol. 2005;
usually difficult. The process of selection requires 286: 3-32. https://doi.org/10.1385/1-59259-827-7:003
multiple subcultures of callus in medium 5. Jhansi RS, Usha R. Transgenic plants: types,
containing the selection pressure and the benefits, public concerns and future. Pharmacy
phytohormone 2,4-D. Multiple subcultures Research. 2013;6:879-883.
resulting in prolonged exposure to auxins such as https://doi.org/10.1016/j.jopr.2013.08.008
2,4-D and the stress of the selection itself can result 6. Larkin PJ, Scowcroft SC. Somaclonal variation- a
in the cells losing their regeneration potential. novel source of variability from cell culture for
Another drawback of this method is the generation plant improvement. Theor. Appl. Genet. 1981; 60:
of pleotropic effects in regenerated plantlets. This 197–214. https://doi.org/10.1007/BF02342540
can only be overcome by obtaining mulitiple 7. Qin-Mei W, Li W. An evolutionary view of plant
somaclonal variants, so as to select a few amongst tissue culture: somaclonal variation and selection.
them having only the desirable traits. More Plant Cell Rep. 2012; 31: 1535-1547.
focused and specific methods of tissue culture and 8. Meins F. Heritable variation in plant cell ann. Rev.
selection that may overcome such road blocks Plant Physiol. 1983; 34:327-46.
need to be developed. 9. Lee M, Phillips RL. The chromosomal basis of
somaclonal variations. An Rev Plant Physiol Plant
Mol Biol. 1988; 39: 413-37.
Conclusion 10. Phillips RL, Kaepppler SM, Peschke VM. Do we
Several resistant somaclonal variant lines have understand somaclonal variation?.Progress in
been generated using the phytopathotoxin in vitro plant cellular and molecular biology. Kluwer Acad.
selection method. This approach is acceptable, Publ, Dordrecht. 1990; 131-141.
simple and has high potential in obtaining 11. Krishna H, Alizadeh M, Singh D, Singh U, Chauhan
desirable variability in crops. Moreover it has the N, Eftekhari M, Sadh RK. Somaclonal variations
potential of generating novel pathways of and their applications in horticultural crops
resistance that can be further exploited in crop improvement. 3 Biotech. 2016; 6: 54. https://doi.org/
improvement. An understanding of the underlying 10.1007/s13205-016-0389-7
molecular pathways to resistance in somaclonal 12. Li R, Bruneau AH, Qu R. Tissue culture-induced
variants is lacking. These pathways need to be morphological somaclonal variation in
explored, and candidate genes identified for st.augustinegrass [Stenotaphrum secundatum
further exploitation in crop improvement. Only by (walt.) kuntze]. Plant Breed. 2010; 129: 96-99.
understanding the many interactions that occur https://doi.org/10.1111/j.1439-0523.2009.01647.
between host plants and pathogens can we utilize 13. Rastogi J, Siddhant, Bubber P, Sharma BL.
cell-culture approach of inducing variations to its Somaclonal variation: a new dimension for
fullest potential. sugarcane improvement. GERF Bulletin of
Biosciences. 2015; 6: 5-10.
14. Egan BT. Chlorotic streak: diseases of sugarcane:
Acknowledgements major diseases edited by Ricaud C, Egan BT,
We acknowledge the financial support under the Gillaspie AG, Hughes CG. Elsevier Science
'Innovative Young Biotechnologist Award' (IYBA) Publication. embryogenesis. Plant Breed. 1989; 121:
grant, Department of Biotechnology (DBT), 269-271.
Government of India to VSA. 15. Moyer JW, Collins WW. Scarlet sweet potato. Hort
Sci. 1983; 18: 111-112.
16. Sarkar AN. Integrated horticulture development in
Authors’ contribution eastern himalayas. M. D Publications. 1994.
VSA compiled and wrote the mini review. SB and 17. Gupta R, Banerjee S, Mallavarapu GR, Sharma S,
SL compiled the Table 1 and the references Khanuja SPS, Shasany AK, Kumar S. Development
section. of a superior somaclone of rose-scented geranium

ISSN: 2348-1900 Horizon e-Publishing Group

52 Plant Science Today (2018) 5(2): 44-54

and a protocol for inducing variants. Hort science. 31. Shepard JF, Bidney D, Shahin E. Potato protoplasts
2002; 37: 632–636. in crop improvement. Science. 1980; 208: 17-24.
https://doi.org/10.1126/science.208.4439.17
18. Arun BA, Joshi K, Chand R, Singh BD. Wheat
somaclonal variants showing earliness, improved 32. Burg HCJ, Ramulu KS, Bredemeijer GMM, Roest S,
spot blotch resistance and higher yield. Euphytica. Dhijkuis P, Hoogen JJV, Houwing A. Patterns of
2003; 132: 235-241. phenotypic and tuber protein variation in plants
https://doi:10.1023/A:1025097224408 derived from protoplast of potato (Solanum
tuberosum l. cv bintje). Plant Sci. 1989; 64: 113-124.
19. Jalaja NC, Sreenivasan TV, Pawar SM, Bhoi PG, https://doi.org/10.1016/0168-9452(89)90157
Garker RM. co 94012- a new sugarcane variety
through somaclonal variation. Sugar Tech. 2006; 33. Ling DH, Vidhyasekharan P, Borromeo ES, Zapata
132-136. https://doi.org/10.1007/BF02943647 FJ, Mew TW. In vitro screening of rice germplasm
for resistance to brown spot disease using
20. Heinz DJ. Sugarcane improvement through Phytotoxin. Theor. Appl. Genet. 1985; 71: 133-135.
induced mutations using vegetative propagules https://doi.org/10.1007/BF00278266
and cell culture techniques. In: induced mutations
in vegetatively propagated plants. ProcPannel 34. Cerato C, Manici LM, Borgatti S, Alicchio R, Ghedini
Vienna, Sept 1972. IAEA pp. 53–59. R, Ghinelli A. Resistance to late blight
https://doi.org/10.17660/ActaHortic.2007.735.19 [Phytophthora infestans (mont.) de bary] of potato
plants regenerated from in vitro selected calli.
21. Carlson PS. Methionine sulfoximine-resistant Potato Research. 1993; 36: 341-351.
mutants of tobacco. Science. 1973; 180: 1366–1368. https://doi.org/10.1007/BF02361801
https://doi.org/10.1126/science.180.4093.1366
35. Venkatachalam P, Rao KS, Kishore PBK,Jayabalan
22. Behnke M. General resistance to late blight of N. Regeneration of late leaf spot resistant
Solanum tuberosum plants regenerated from callus groundnut plants from Carcosporidium
resistant to culture filtrates of Phytophthora personatum culture filtrate treated callus. Curr. Sci.
infestans. Theor. Appl. Genet. 1980; 56: 151-152. 1998; 74: 61-65.
https://doi.org/10.1007/BF00286676 http://www.jstor.org/stable/24100665
23. Lestari EG. In vitro selection and somaclonal 36. Rao S, Basavaraj K, Kaviraj CP. In vitro selection of
variation for biotic and abiotic stress tolerance. pigeon pea cell lines and regeneration of plantlets
Biodiversitas. 2006; 7: 297–301. from tolerant callus to culture filtrate of Fusarium
https://doi.org/10.13057/biodiv/d070320 odum Buttler. Plant Cell Biotech. Mol Bio. 2006;
7(1&2): 69-72.
24. Dehgahi R, Subramaniam S, Zakaria L, Joniyas A.
Review of research on in vitro selection of 37. Thakur M, Sharma D, Sharma S. In vitro selection
Dendrobium sonia-28, against Fusarium and regeneration of carnation (Dianthus
proliferatum. Int. J. Sci. Res. in Agricultural cayophyllus l.) plants resistant to culture filtrate of
Sciences. 2016; 3: 050-061. Fusarium oxysporum f. sp. dianthi. Plant Cell Rep.
http://dx.doi.org/10.12983/ijsras 2002; 20: 825–828. https://doi.org/10.1007/s00299-
001-0412-1
25. Kintzios S, Koliopoulos A, Karyoti E, Drossopoulos J,
Holevas CD, Grigoriu A, Panagopoulos. In vitro 38. Saxena G, Verma PC, Rahman L, Banerjee S, Shukla
reaction of sunflower (Helianthus annus l.) to the RS, Kumar S. Selection of leaf blight-resistant
toxin (s) produced by Alternaria alternata, the Pelargonium graveolens plants regenerated from
causal agent of brown leaf spot. Phytopathol. 1996; callus resistant to a culture filtrate of Alternaria
144: 465–470. https://doi.org/10.1111/j.1439- alternate. Crop Protection. 2008; 27: 558–565.
0434.1996.tb00325 https://doi.org/10.1016/j.cropro.2007.08.013
39. Rao S, Ramgopal S. Effect of Alternaria helianthi
26. Thanutong P, Furusawa I, Yamamoto M. Resistant culture filtrate on callus and regeneration of
tobacco plants from protoplast derived calluses for plantlets from tolerant callus in sunflower
their resistance to pseudomonas and alternaria (Helianthus annuus l.). Indian Journal of
toxins. Theor. Appl. Genet. 1983; 66: 209–215. Biotechnology. 2010;9: 187: 191.
https://doi.org/10.1007/BF00251145 http://nopr.niscair.res.in/handle/123456789/7798
27. Penna S, Vitthal SB, Yadav PV. In vitro mutagenesis 40. Bhardwaj SV, Thakur T, Sharma R, Sharma P. In
and selection in plant cultures and their prospects vitro selection of resistant mutants of ginger
for crop improvement. Bioremediation, (Zingiber officinalerosc.) against wilt pathogen
Biodiversity and Bioavailability. 2012; 6: 6-14. (Fusarium oxysporum f. sp. Zingiber itrujillo). Plant
28. Gengenbach BG, Green CE, Donovan CM. Dis Res. 2012; 27: 194–199.
Inheritance of selected pathotoxin resistance in 41. Gostimsky SA, Kokaeva ZG, Konovalov FA.
maize plants regenerated from cell cultures. Proc Studying plant genome variation using molecular
Natl. Acad. Sci. 1977; 74: 5113-5117. markers. Russ J Genet. 2005; 41:3 78–88.
29. Behnke M. Selection of potato callus for resistance https://doi.org/10.1007/s11177-005-0101-1
to culture filtrates of Phytophthora infestans and 42. Nayak S, Debata BK, Srivastava VK, Sangwan NS.
regeneration of resistant plants. Theor. Appl. Evaluation of agronomically useful somaclonal
Genet. 1979; 55: 69-71. variants in jamrosa (a hybrid cymbopogon) and
https://doi.org/10.1007/BF00285192 detection of genetic changes through RAPD. Plant
30. Shahin EA, Spivey R. A single dominant gene for Sci. 2003; 164: 1029–1035.
fusarium wilt resistance in protoplast-derived https://doi.org/10.1016/S0168-9452(03)00090-6
tomato plants. Theor. Appl. Genet. 1986; 73: 164- 43. Kumar N, Modi AR, Singh AS, Gajera BB, Patel AR,
169. https://doi.org/10.1007/BF00289270 Patel MP, Subhash N. Assessment of genetic fidelity

Horizon e-Publishing Group ISSN: 2348-1900

Plant Science Today (2018) 5(2): 44-54 53

of micropropagated date palm Phoenix dactylifera entomologica Hungarica 2003;38:21-28.

L.) plants by RAPD and ISSR markers assay. 58. Behnke M. Selection of dihaploid potato callus for
Physiol. Mol Biol. 2010; 6: 207‐213. resistance to culture filtrate of Fusarium
https://doi.org/10.1007/s12298-010-0023-9 oxysporum. Pflanzenzuchtg. 1980; 85: 254-258.
44. Saravanan S, Sarvesan R, Vinod MS. Identification 59. Jayasankar S, Litz RE. Characterization of
of DNA elements in somaclonal variants of embryogenic mango cultures selected for
Rauvolfia serpentine (L.) arising from indirect resistance to Colletotrichum gloeosporioides culture
organogenesis as evaluated by ISSR analysis. filtrate and phytotoxin. Theor. Appl. Genet. 1998;
Indian J. of Sci. and Tech. 2011; 4: 1241-1245. 96: 823-831. https://doi.org/10.1007/s001220050808
45. Bairu MW, Aremu AO, Staden JV. Somaclonal 60. Deng ZN, Gentile A, Domina F, Nicolosi E, Tribulato
variation in plants: causes and detection methods. E. Selecting lemon protoplasts for insensitivity to
Plant Growth Regul. 2011; 63: 147–173. Phoma tracheiphila toxin and regenerating tolerant
https://doi.org/10.1007/s10725-010-9554-x plants. J. Am. Soc.Hortic Sci. 1995; 120: 902–905.
46. Sharma NK, Skidmore DI. In vitro expression of 61. Zhang LQ, Cheng ZH, Khan MA, Zhou YL. In vitro
partial resistance to Phytophthora palmivora by selection of resistant mutant garlic lines by using
shoot cultures of papaya. Plant Cell Tiss. Org. 1988; crude pathogen culture filtrate of Sclerotium
14: 187-196. https://doi.org/10.1007/BF00043409 cepivorum australas. Plant Pathol. 2012; 41: 211–
47. Song HS, Lim SM, Widholm JM. Selection and 217. https://doi.org/10.1007/s13313-011-0109-z
regeneration of soybeans resistant to the 62. Devarumath RM, Nandy S, Rani V, Marimuthu S,
pathotoxic culture filtrates of Septoria glycines. Muraleedharan N, Raina SN. RAPD, ISSR and RFLP
Phytopathol. 1977; 84: 948-951. fingerprints as useful markers to evaluate genetic
48. Ganesan M, Jayabalan N. Isolation of disease intergrity of diploid and triploid elite tea clones
tolerant cotton (Gossypium hirsutuml. cv. svpr 2) representing Camellia sinensis (China type) and C.
plants by screening somatic embryos with fungal assamica ssp. assamica (Assam type). Plant Cell
culture filtrate. Plant Cell Tiss. Org. 2006; 87: 273- Rep. 2002; 21: 166–173.
284. http://www.jstor.org/stable/41998136 63. Orbovic V, Syvertsen JP, Bright D, Clief VDL,
Graham JH. Growth of citrus seedlings and their
49. Chawla HS, Wenzel G. In vitro selection of barley susceptibility to Phytophthora root rot are affected
and wheat for resistance against
by PO3 and PO4 sources of phosphorus. Plant Nutr.
Helminthosporium sativum. Theor. Appl. Genet.
2008; 31: 774–787.
1987; 74: 841-845. https://doi.org/10.1007/BF00247566
64. Sanchez TLF, Quiroz FF, Loyola VV, Infante D.
50. Fujime F, Fujime G. Use of culture filtrates of
Culture-induced variation in plants of Coffea
Pyrenochaeta lycopersici in tests for selecting
arabica cv. Caturrarojo, regenerated by direct and
tolerant varieties of tomato. J. Plant Pathol. 2003;
indirect somatic embryogenesis. Mol Biotechnol.
85: 131-133. http://www.jstor.org/stable/41998136
2003; 23: 107–115.
51. Hartman CL, Mccoy TJ, Knous TR. Selection of 65. Bhattacharya S, Dey T, Bandopadhyay T, Ghosh P.
alfalfa (Medicago sativa) cell lines and regeneration Genetic polymorphism analysis of somatic embryo-
of plants resistant to the toxin(s) produced by derived plantlets of Cymbopogon flexuosus through
Fusarium oxysporum f. sp. medicaginis. Plant RAPD assay. Plant Biotechnol Rep. 2008; 2: 245–252.
Sci.Lett. 1984; 34: 183-219. 66. Biswas MK, Dutt M, Roy UK, Islam R, Hossain M.
https://doi.org/10.1016/0304-4211(84)90141 Development and evaluation of in vitro somaclonal
52. McCoy TJ. Tissue culture selection for disease variation in strawberry for improved horticultural
resistant plants. Iowa State J. Res. 1988; 62: 503-521. traits. Sci. Hort. 2009; 122: 409–416.
53. Vidysekaran P, Ling DH, Borromeo ES, Zapata FJ, 67. Gesteira AS, Otoni WC, Barros EG, Moreira MA.
Mew TW. Selection of brown spot- resistant rice RAPD – based detection of genomic instability in
plant from Helminthosporium oryzae toxin- soybean plants derived from somatic. Plant Breed.
resistant cultures. Annals of Applied Biol. 1990; 2002; 121: 269-271. https://doi.org/ 10.1046/j.1439-
117: 515-523. https://doi.org/ 10.1111/j.1744- 0523.2002.00708.x
7348.1990.tb04818 68. Jin S, Mushke R, Zhu H, Tu L, Lin Z, Zhang Y, Zhang
54. Vos JE, Berjak MHS, Watt MP, Toerien AJ. In vitro X. Detection of somaclonal variation of cotton
selection and commercial release of guava wilt (Gossypium hirsutum) using cytogenetics, flow
resistant rootstocks. Acta. Horticulturae. 1998; 513: cytometry and molecular markers. Plant Cell Rep.
69-79. 2008; 27: 1303–1316.
https://doi.org/10.17660/ActaHortic.1998.513.7
69. Li X, Yu X, Wang N, Feng Q, Dong Z, Liu L, Shen J,
55. Bajpai A, Chandra R, Mishra M, Tiwari RK. Liu B. Genetic and Epigenetic instabilities induced
Regenerating Psidium spp. for screening wilt by tissue culture in wild barely (Hordeum
resistance rootstock under in vitro conditions. Acta brevisubulatum (Trin) Link). Plant Cell, Tissue and
Horticulturae. 2007; 735: 145-154. Organ culture. 2007; 90: 153-168.
56. Maier FJ, Oettler G. Selection for the fusarium toxin 70. Campbell B, LeMare S, Piperidis G, Godwin I. IRAP,
deoxynivalenol in callus culture of triticale. In a retrotransposon-based marker system for the
Third European Fusarium Seminar, IHARR, Poland. detection of somaclonal variation in barley. Mol
1992; 43-49. Breed. 2010; 27: 103-206.
57. Mohanraj D, Padmanaban P, Karunakaran M. https://doi.org/10.1007/s11032-010-9422-4
Effect of pytotoxin of Colletrichum falcatum went. 71. Sheidai M, Aminpoor H, Noormohammadi Z, Farahani
(Physalophora tucumanensis) on sugarcane in F. Genetic variation induced by tissue culture in
tissue culture. Acta. phytopathologicaet.

ISSN: 2348-1900 Horizon e-Publishing Group

54 Plant Science Today (2018) 5(2): 44-54

Banana (Musa acuminate L.) cultivar Cavandish Dwarf 79. Heinz DJ, Mee GWP. Morphologic, cytogenetic and
Masoud. Geneconserve. 2009; 9: 1-10. enzymatic variation in Saccharum species hybrid
72. James AC, Pereza ES, Herrera VA, Martinez O. clones derived from callus tissue. Amer J. Bot. 1971;
Application of the amplified fragment length 58: 257-262
polymorphism (AFLP) and the methylation- 80. De la PR, Gonza´lez A, Ruiz M, Polanco C.
sensitive amplification polymorphism (MSAP) Somaclonal variation in rye (Secalecereale L.)
techniques for the detection of DNA polymorphism analyzed using polymorphic and sequenced AFLP
and changes in DNA methylation in markers. In vitro Cell DevBiol Plant. 2008; 44: 419–
micropropagated bananas. In: Jain SM, Swennen R 426.
(eds) Banana improvement: cellular, molecular 81. Aversano R, Savarese S, Nova JMD, Frusciante L,
biology, and induced mutations. Science Punzo M, Carputo D. Genetic stability at nuclear
Publishers. 2004; 287–306. and plastid DNA level in regenerated plants of
73. Asif MJ, Othman RY. Characterization of Fusarium Solanum species and hybrids. Euphytica. 2009;
wilt-resistant and fusarium wilt-susceptible 165: 353–361. https://doi.org/ 10.1007/s10681-008-
somaclones of banana cultivar rastali (Musa AAB) 9797-z
by random amplified polymorphic DNA and 82. Albani MC, Wilkinson MJ. Inter simple sequence
retrotransposon markers. Plant Mol Biol. Rep. repeat polymerase chain reaction for the detection
2005; 23: 241–249. of somaclonal variation. Plant Breed. 1998; 117:
74. Ghag SB, Shekhawat UKS, Ganapathi TR. 573–575.
Characterization of fusarium wilt resistant 83. Sharma S, Bryan G, Winfield M, Millam S. Stability
somaclonal variants of banana cv. rasthali by of potato (Solanum tuberosum L.) plants
cDNA-RAPD. MolBiol Rep. 2014; 41(12): 7929–7935. regenerated via somatic embryos, axillary bud
https://doi.org/10.1007/s11033-014-3687-3 proliferated shoots, microtubers and true potato
75. Sultana R, Tahira F, Tayyab H, Khurram B, Shiekh seeds: a comparative phenotypic, cytogenetic and
R. RAPD characterization of somaclonal variation molecular assessment. Planta. 2007; 226: 1449–
in indica basmati rice. Pak J Bot. 2005; 37: 249–262. 1458.
76. Ngezahayo F, Dong Y, Liu B. Somaclonal variation 84. Zhang M, Wang H, Dong Z, Qi B, Xu K, Liu B. Tissue
at the nucleotide sequence level in rice (Oryza culture induced variation at simple sequence
sativa L.) as revealed by RAPD and ISSR markers, repeats in sorghum (Sorghum bicolor L.) is
and by pairwise sequence analysis.Theor.Appl genotype-dependent and associated with down-
Genet. 2007; 48: 329–336. regulated expression of a mismatch repair gene,
77. Hsu TW, CTsai WC, Wang DP, Lin S, Hsiao YY, Chen MLH3. Plant Cell Rep. 2010; 29: 51–59.
WH, Chen HH. Differential gene expression 85. Rodrı´guez LC, Bravo H, Wetten A, Wilkinson M.
analysis by cDNAAFLP between flower buds of Detection of somaclonal variation during cocoa
Phalaenopsis Hsiang Fei cv. H. F. and its somatic embryogenesis characterised using
somaclonal variant. Plant Sci. 2008; 175: 415–422. cleaved amplified polymorphic sequence and the
78. Kuznetsova OI, Ash OA, Hartina GA, Gostimskij SA. new freeware Artbio. Mol Breeding. 2010; 25: 501–
RAPD and ISSR analyses of regenerated pea Pisum 516.
sativum L. plants. Russ J. Genet. 2005; 41: 60–65.

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