Development of new microsatellite markers for DNA fingerprinting pattern of black gram (Vigna mungo L. Hepper) and green gram (Vigna radiate L. Wilckzek)

Abstract

SSR markers are considered to be the most ideal marker for genetic studies because they are multi-allelic, abundant, randomly and widely distributed throughout the genome, co-dominant that could differentiate plants with homozygous or heterozygous alleles, simple to assay, highly reliable, reproducible. Microsatellite markers are highly polymorphic and informative and could be successfully used for genome analysis in black gram & green gram. Microsatellite markers were used to evaluate genetic diversity in 17 indigenous cultivars of pulse crops (11 cultivars of green gram and 6 cultivars of black gram respectively). They are subjected to variability analysis with 26 microsatellite markers for identification efficient primers to conclude the nature of molecular diversity present among the pulses. The SSR primer G228 showed 63.63% of polymorphism followed by MB-SSR 238 (45.45%) and G006 (36.36%). The 12 microsatellite markers produced 15.90 % polymorphism with banding ranged up to 7 with an average of 2.3 polymorphic banding patterns per SSR primer. Similarly for black gram, three random microsatellite primers G006 (50%) and G166 and G204 (33.33%) revealed considerable DNA polymorphism. The 14 random SSR primers produced 8.33% of polymorphism with banding ranged up to three with an average of 1.28 polymorphic banding pattern per SSR primer. The Distinguish Power (D), Polymorphism Information Content (PIC) value and Marker Index (MI) values revealed some SSR primers like G006, G204 and G166 can alone amplified distinct banding pattern, where as a combination of (G228+G006), (G228+G304) for green gram and the combination (G006+G166) can be used for black gram for ascertaining genetic diversity at any stage of crop growth period for green gram or black gram. From the present study we can conclude that selective microsatellite markers are highly polymorphic, informative and easily reproducible, which can be successfully used either as single or with combination for molecular characterization of crop species belonging to Vigna species.