ANSHU BAJAJ AND SURENDER K. PAHUJA
Department of Biotechnology & Molecular Biology, CCS HAU, Hisar-125 004 (Haryana), India
Department of Genetics & Plant Breeding, CCS HAU, Hisar-125 004 (Haryana), India
*(e-mail: pahujask66@gmail.com)
(Received: 20 September 2024; Accepted: 30 September 2024)
SUMMARY
Cyamopsis tetragonoloba (L.) Taub, commonly known as guar belongs to family Fabaceae. Guar seed has an important place in industry because of its galactomannan rich endosperm. Good quality genomic DNA was isolated from 48 independent cluster bean varieties and conditions were optimized for further amplification by PCR using random one hundred and thirty decamer operon primers. An evaluation was made for the application of RAPD as a genetic marker system in commercially important cluster bean varieties and a dendrogram was prepared. Various factors influencing the RAPD amplification were optimized and the data revealed that 50ng of template DNA, 1.5mM Mg2+ ion concentration, 4.0 U/reaction of Taq DNA polymerase and annealing temperature of 400C was found to be essential for reproducible RAPD banding pattern. Cluster bean cultivars revealed significant polymorphism with reference to RAPD markers showing authentic genotypic diversity among its races. Out of a total of one hundred thirty operon primers employed; ninety-seven random primers showed amplified products with 91.34% polymorphism, yielding 644 polymorphic and 61 monomorphic alleles. All the genotypes could be grouped in two major and three minor sub-clusters, when binary matrix was subjected to NTSYS-pc software analysis and clustered dendrogram was constructed. The results of the present study can be used for molecular breeding and improvement of cluster bean for various desired traits through hybridization in future.
Key words: Guar, Cyamopsis tetragonoloba, DNA extraction, molecular markers, RAPD, PCR