Induced mutagenesis in Jatropha curcas L. using ethyl methanesulphonate (EMS) and assessment of DNA polymorphism through RAPD markers

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.