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Inter- and Intra-Specific Genetic Diversity Analysis in Thirty Genotypes of Withania Somnifera (Ashwagandha) using Protein Profile Pattern

Sunita Khatak, Om Prakash Yadav and Ramender Kaith

Thirty genotypes of Withania somnifera (L) Dunal also known as Ashwagandha were collected from three different states of Indi a and diversity analysis was carried out using electrophoregrams using gradient gels (9-15%) from four to five week old leaves. A total of 11 bands were generated having Rm (Relative mobility) value ranging from 0.05 to 0.65 and of M. wt. in range of < 6.5 kDa to 45 kDa. The electrophoregram obtained using SDS-PAGE were used to estimate genetic similarity matrix, which ranged from 0.18 to 1.0 indicating high variability. The average similarity index value was 0.569. The dendrogram constructed from SDS-PAGE revealed one major cluster comprising all genotypes at a similarity coefficient of 0.55, which further bifurcates into two distinct clusters. Distinct clustering of genotypes of particular location was not observed although protein profile as general was considered as biochemical fingerprints of particular genotypes. Unique bands were observed using protein profile analysis distinguishing particular genotypes from rest. A unique band of Rm value 0.56 and 0.65 of molecular weight less than 6.5 kDa were found to be present in all the genotypes from Mandsaur district (M.P.) and Haryana along with two genotypes WS-223 and WS-224 from Udaipur (Rajasthan), while rest of the genotypes lack this specific band. Genotype WS-124 was clearly distinguished showing presence of a single band with Rm value (0.21) of molecular weight 22.3 kDa. The genotypes collected from different locations did not form well defined distinct clusters and were interspersed with each other.