Molecular characterization of Vigna radiata (L.) Wilczek genotypes based on nuclear ribosomal DNA and RAPD polymorphism

Abstract

Mungbean germplasm characterization, evaluation and improvement are fundamentally based on morpho-agronomic traits. The lack of break-through in mungbean production has been due to non-availability of genetic variability for high yield potential. Forty-four genotypes of mungbean [Vigna radiata (L.)Wilczek] were subjected to random amplified polymorphic DNA (RAPD) analysis to assess the genetic diversity and relationships among the genotypes. Multilocus genotyping by twelve RAPD primers generated 166 markers and detected an average of intraspecific variation amounting to 82% polymorphism in banding patterns. Dendrogram obtained from cluster analysis delineated all the 44 genotypes into six clusters. Higher values of Nei's gene diversity (h) and Shannon information index (i) and genetic distance analysis validate existence of wide genetic diversity among mungbean genotypes tested. Besides internal transcribed spacer (ITS) length variations, single nucleotide polymorphisms (SNPs) and insertions/deletions (INDELS) were detected at number of sites in nuclear rDNA region and the sequences of representatives of each sub-cluster and all distinct genotypes have been submitted to NCBI database and assigned Gen accession numbers HQ 148136-148147. Multiple sequence alignment revealed further lineages of distinct genotypes with main RAPD clusters. The measures of relative genetic distances among the genotypes of mungbean did not completely correlate the geographical places of their development. The homogeneous phenotypic markers proved insufficient in exhibiting genetic divergence among mungbean genotypes studied. RMG-62, RMG-976, and NDM-56 have been identified as potential source of parents for crop improvement. RAPD primers, OPA-9 and OPA-2 as polymorphic genetic markers and number of pods/plant and number of seeds/plant as dependable phenotypic markers have been identified for improving yield potentials. This genetic diversity will be of significance in developing intraspecific crosses in mungbean crop improvement programme.

Application of ISSR markers for verification of F1 hybrids in mungbean (Vigna radiata)

“…Several molecular markers have been used in mungbean, including amplified fragment length polymorphism (AFLP), restriction fragment length polymorphism (RFLP), random amplified polymorphic DNA (RAPD), simple sequence repeat (SSR), and inter-simple sequence repeats (ISSR) (Chaitieng et al, 2002;Afzal et al, 2004;Tangphatsornruang et al, 2009;Tantasawat et al, 2010b;Raturi et al, 2012). Afzal et al (2004) have used RAPD to evaluate the genetic diversity of 21 cultivated mungbeans from Thailand, Bangladesh, and India.…”

Genetic Confirmation of Mungbean (Vigna radiata) and Mashbean (Vigna mungo) Interspecific Recombinants using Molecular Markers

“…Many other studies reported that RAPD markers are efficient in amplifying DNA from dispersed polymorphic loci from the genome ( Baral and Bosland, 2002 ; Rasul et al, 2007 ; Wang et al, 2011 ). These markers can be used to assess genetic variability among and between Vigna species ( Kaga et al, 1996 ; Santalla et al, 1998 ; Lakhanpaul et al, 2000 ; Ba et al, 2004 ; Saini et al, 2004 ; Raturi et al, 2012 ). Wang et al (2008) reported thirty gene derived markers which were employed to reveal interspecific phylogenetic relationships and genetic diversity among 48 accessions of 12 Vigna species.…”

“…Molecular markers have been used for various purposes including determination of genetic relationships between individuals, construction of linkage maps, population genetics, phylogenetic studies, mapping of useful genes, and marker assisted selection/backcrosses. Several molecular marker systems, including randomly amplified polymorphic DNA (RAPD), universal rice primers (URP), simple sequence repeats (SSR), amplified fragment length polymorphism (AFLP), restriction fragment length polymorphism (RFLP), and inter-simple sequence repeats (ISSR) have been used in mungbean to characterize DNA variation patterns within and among closely related species ( Dikshit et al, 2007 ; Raturi et al, 2012 ). DNA markers allow rapid identification of cultivars, hybrids, somaclonal variants, and clones with high efficiency and less labor cost ( Reddy et al, 2002 ).…”

Mungbean yellow mosaic virus (MYMV): a threat to green gram (Vigna radiata) production in Asia

“…By determining the allele of a DNA marker, plants that possess particular genes or quantitative trait loci (QTLs) may be identified based on their genotype rather than their phenotype. Different markers are used to study genetic diversity (Chattopadhyay et al 2005;Roopa et al 2008;Zhao et al 2010;Datta et al 2012;Raturi et al 2012;Zhong et al 2012) and tag the MYMV resistance genes in mungbean (Selvi et al 2006;Chen et al 2012;Karthikeyan et al 2012;Dhole & Kanda 2013;Sudha et al 2013a). MAS, to a great extent, has improved the efficiency of resistance breeding in MYMV and has shown some significant success too.…”