Assessment of genetic diversity, population structure and sex identification in dioecious crop, Trichosanthes dioica employing ISSR, SCoT and SRAP markers

Abstract

Twenty inter-simple sequence repeat (ISSR) and twenty two start codon targeted (SCoT) primers were employed to analyze genetic diversity and population structure among 52 Trichosanthes dioica Roxb. accessions collected from nine different eco-geographical regions of India. ISSR markers proved to be more informative in genetic diversity assessment and produced higher mean number of polymorphic bands (15.25 with 95.96% polymorphism) and polymorphic information content (PIC) value (0.47) compared to SCoT markers (12.55 polymorphic bands with 92.20% polymorphism and PIC: 0.45). Total genetic diversity (Ht) and genetic diversity within populations (Hs) in T. dioica accessions was found to be very high (0.45 and 0.43, respectively). AMOVA analysis also revealed higher genetic variation within populations (81%) than among them (19%). Among different T. dioica populations, very low genetic differentiation (Gst: 0.05) and high gene flow (Nm: 9.32) were observed. T. dioica populations of Bihar state were found to be highly diverse and Kolkata and Cuttack populations were least diverse. T. dioica male plants were more variable than females. UPGMA, Neighbor-Joining and population structure analyses divided T. dioica populations into three main clusters. First cluster comprised of Meerut population, second cluster included of Cuttack and Kolkata populations and populations of Bihar, Delhi and Kanpur occurred in third cluster. Genetic diversity was found to be strongly positively correlated with the latitude and strongly negatively correlated with annual mean rainfall of different T. dioica cultivated regions. For sex identification, one SRAP primer combination, 'Em-6/Me-4' amplified two molecular markers of around 230 and 290 bp specific to male T. dioica plants of Bihar, Kanpur, North Delhi and Meerut populations and were completely absent from female plants.