Abstract
Ethyl methanesulphonate (EMS) has been employed in a number of genotoxic studies in plants as a model alkylated agent that readily reacts with DNA-producing alkylated nucleotides. Therefore, the present study was aimed at assessing DNA polymorphism induced by different concentrations (control, 1, 2, 3, and 4%) of EMS through a Randomly Amplified Polymorphic DNA (RAPD) marker analysis. The improved agronomic traits such as germination, flowering, maturity, seed traits, and oil content were recorded in 1% EMS-treated plants, while the corresponding parameters reduced significantly (P > 0.05) in 4% EMS-treated plants as compared to the control. Among 25 random primers used, 19 primers produced polymorphic bands. The number of amplicons varied from 1 to 8 with an average of 3.68 bands, of which 2.12 were polymorphic. The highest polymorphic bands (6) and percentage of polymorphism (85.71) were produced by the primer OPAK-20. In a dendrogram constructed based on Jaccard’s coefficient similarity, the treated plants and control were grouped into three clusters: (a) control and 2 and 3% concentrations of EMS-treated plants merged together; (b) 1% concentration of EMS-treated plants clustered alone; (c) 4%concentration of EMS-treated plants also clustered alone. We conclude that the effect of EMS could change the pattern of germination, flowering, seed yield, and oil content of J. curcas. DNA polymorphism detected by RAPD marker analysis offered a useful biomarker assay for the evaluation of effects of chemical mutagens.
Similar content being viewed by others
References
Atienzar FA, Cordi B, Donkin ME, Evenden AJ, Jha AN, Depledge MH. 2000. Comparison of ultraviolet-induced genotoxicity detected by random amplified polymorphic DNA with chlorophyll fluorescence and growth in a marine macroalgae, Palmaria palmate. Aquat. Toxicol. 50: 1–12
Belaj A, Trujilo I, Rosa R, Rallo L, Gimenez MJ. 2001. Polymorphism and discrimination capacity of randomly amplified polymorphic markers in an olive germplasm bank. J. Am. Soc. Hort. Sci. 126: 64–71
Bhagwat A, Krishna TG, Bhatia CR. 1997. RAPD analysis of induced mutants of groundnut (Arachis hypogaea L.). J. Genet. 76: 201–208
Carlier JD, Reis A, Duval MF, Conppens G, Leitao JM. 2004. Genetic maps of RAPD, AFLP, and ISSR markers in Ananas bracteatus and Ananas comosus using the pseudo-testcross strategy. Plant Breed. 123: 186–192
Cenkci S, Cigerci IH, Yildiz M, Ozay C, Bozdag A, Terzi H. 2010. Lead contamination reduces chlorophyll biosynthesis and genomic template stability in Brassica rapa L. Environ. Exp. Bot. 67: 467–473
Chandra A, Tewari S. 2007. Isolation of genomic DNA from Stylo Species without liquid nitrogen suitable for RAPD and STS analyses. Cytologia 72: 287–297
Chaparro E, Werner DJ, O’Malley D, Sederoff RR. 1994. Targeted mapping and linkage analysis of morphological isozyme, and RAPD markers in peach. Theor. Appl. Genet. 83: 194–200
Charbaji T, Nabulsi I. 1999. Effect of low doses of gamma irradiation on in vitro growth of grapevine. Plant Cell Tiss. Org. Cult. 57: 129–132
Demeke T, Adams RP, Chibbar R. 1992. Potential taxonomic use of random amplified polymorphic DNA (RAPD) — a case study in study. Theor. Appl. Genet. 84: 990–994
Deshwall RPS, Singh R, Malik K, Randhawa GJ. 2005. Assessment of genetic diversity and genetic relationships among 29 populations of Azadirachta indica A. Juss. using RAPD markers. Genet. Resour. Crop Evol. 52: 285–292
Doyle JJ, Doyle JL. 1990. Isolation of DNA from fresh plant tissue. Focus 12: 12–15
Erdem G, Oldacay S. 2004. Employment of RAPD technique to assess the genetic stability of Helianthus annuus treated with different mutagenic agents. J. Appl. Sci. 4: 277–281
Ganesh-Ram S, Parthiban TK, Senthilkumar R, Thiruvengadam V, Paramathma M. 2008. Genetic diversity among Jatropha species as revealed by RAPD markers. Genet. Resour. Crop Evol. 55: 803–809
Gichner T. 2003. Differential genotoxicity of ethyl methanesulphonate, N-ethyl-N nitrosourea and maleic hydrazide in tobacco seedlings based on data of the comet assay and two recombination assays. Mutat. Res. 538: 171–179
Green AG, Marshall DR. 1984. Isolation of induced mutants in linseed (Linum usitatissimum) having reduced linolenic acid content. Euphytica 33: 321–328
Gupta S, Srivastava M, Mishra GP, Naik PK, Chauhan RS. 2008. Analogy of ISSR and RAPD markers for comparative analysis of genetic diversity among different Jatropha curcas genotypes. Afr. J. Biotechnol. 7: 4230–4243
Ikbal KSB, Dhillon RS. 2010. Evaluation of genetic diversity in Jatropha curcas L. using RAPD markers. Indian J. Biotechnol. 9: 50–57
Ito A, Hayama H, Kashimura Y, Yoshioka H. 2001. Effect of maleic hydrazide on endogenous cytokinin contents in lateral buds, and its possible role in flower bud formation on the Japanese pear shoot. Sci. Hortic. 87: 199–205
Khan IA. 1984. Mutations induced by gamma-irradiation, ethyl methane sulfonate and hydrazine hydrate in mungbean (Phaseolus aureus Roxb.). Bot. Bull. Acad. Sinica 25: 103–110
Khan S, Goyal S. 2009. Improvement of mungbean varieties through induced mutations. Afr. J. Plant Sci. 3: 174–180
King AJ, He W, Cuevas JA, Freudenberger M, Ramiaramanana D, Graham IA. 2009. Potential of Jatropha curcas as a source of renewable oil and animal feed. J. Exp. Bot. 60: 2897–2905
Liu W, Li P, Qi X, Zhou Q, Sun T, Yang Y. 2005. DNA changes in barley (Hordeum vulgare) seedlings induced by cadmium pollution using RAPD analysis. Chemosphere 61: 158–167
Liu W, Yang YS, Zhou Q, Xie L, Li P, Sun T. 2007. Impact assessment of cadmium contamination on rice (Oryza sativa L.) seedlings at molecular and population levels using multiple biomarkers. Chemosphere 67: 1155–1163
Luo S, He P, Zheng X, Zhou P. 2002. Inheritance of RAPD markers in an interspecific F1 hybrid of grape between Vitis quinquangularis and V. vinifera. Sci. Hortic. 93: 19–28
Lyons-Alcantara M, Mooney R, Lyng F, Cottell D, Mothersill C. 1998. The effect of cadmium exposure on the cytology and function of primary cultures from rainbow trout. Cell Biochem. Funct. 16: 1–13
Micke A, Maluszynski M, Donini B. 1987. Induced mutations for crop improvement — a review. Trop. Agric. 64: 259–278
Mishra RC, Mohapatra BD, Panda BS. 2001. High yielding mutants of black gram variety PH-25. Mutat. Breed. Newslett. 45: 39
Naik PM, Murthy HN. 2009. The effects of gamma and ethyl methanesulphonate treatments on agronomical traits of Niger (Guizotia abyssinica Cass.). Afr. J. Biotech. 8: 4459–4464
Nichterlein K, Marquard R, Friedt W. 1988. Breeding for modified acid composition by induced mutations in linseed (Linum usitatissimum L.). Plant Breed. 101: 190–199
Ong TM, Song B, Qian HW, Wu ZL, Whong WZ. 1998. Detection of genomic instability in lung cancer tissues by random amplified polymorphic DNA analysis. Carcinogenesis 19: 233–235
Pandey RK, Datta SK. 1995. Gamma ray induced cotyledonary variabilities in Jatropha curcas L. J. Nucl. Agric. Biol. 6: 24–62
Punia MS. 2007. Current status of research and development on Jatropha (Jatropha curcas) for sustainable biofuel production in India. In: USDA Global Conference on Agricultural Biofuels: Research and Economics, Minneopolis, Minnesota, pp 20–22
Rowland GG. 1990. An EMS-induced low-linolenic-acid mutant in McGregor flax (Linum usitatissimum L.). Can. J. Plant Sci. 71: 393–396
Sestras R. 2004. Horticultural plant breeding (in Romanian), Ed, Academic Press, Cluj-Napoca pp 334
Sharma R, Mahla HR, Mohpatra T, Bhargva SC, Sharma MM. 2003. Isolating plant genomic DNA without liquid nitrogen. Plant Mol. Biol. Rep. 21: 43–50
Sneath PHA, Sokal RR. 1973. Numerical Taxonomy: The principles and practice of numerical classification, San Francisco, Freeman, pp 573
Uma S, Sudha S, Saraswathi MS, Nickavasagam M, Selvarajam R, Durai P, Sathiamoorthy S, Siva AS. 2004. Analysis of genetic diversity and phylogenetic relationships among indigenous and exotic Silk (AAB) group of bananas using RAPD markers. J. Hortic. Sci. Biotech.79: 523–527
Venkatachalam P, Geetha N, Jayabalan N. 1999. Twelve new groundnut (Arachis hypogaea L.) mutated germplasm registered in ICRISAT gene bank. Mutat. Breed. Newslett. 44: 15–17
Werner T, Motyka V, Strnad M, Schmulling T. 2001. Regulation of plant growth by cytokinin. J. Plant Biol. 98: 10487–10492
Zhiyi R, Haowen Y. 2004. A method for genotoxicity detection using random amplified polymorphism DNA with Danio rerio. Ecotoxicol. Environ. Safety 58: 96–103
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Dhakshanamoorthy, D., Selvaraj, R. & Chidambaram, A. Induced mutagenesis in Jatropha curcas L. using ethyl methanesulphonate (EMS) and assessment of DNA polymorphism through RAPD markers. J. Crop Sci. Biotechnol. 16, 201–207 (2013). https://doi.org/10.1007/s12892-012-0079-x
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s12892-012-0079-x