Abstract

Genetic diversity of 48 sea buckthorn genotypes collected from 16 locations in four diverse geographical regions of Himachal Pradesh and Jammu and Kashmir states of north-western Himalayan region of India, was analyzed using random amplified polymorphic DNA (RAPD), inter-simple sequence repeat (ISSR), simple sequence repeat (SSR) and MADS-box gene markers. While 21 decamer primers generated 155 loci, of which 148 (95.48%) were polymorphic with average of 7.3 loci per primer, ten ISSR primers generated 86 loci (8.6 loci per primer), out of which 83 (96.51%) were polymorphic. Six SSRs generated 18 loci (3 loci per primer), of which 17 (94.45%) were polymorphic, whereas 3 SSR primers resulted into monomorphic bands. Two MADS-box primers on the other hand generated 13 loci (6.5 loci per primers) with 100% polymorphism. RAPD, ISSR, SSR and MADS box gene-specific primers generated amplicons with polymorphism information content (PIC) values of 0.60 to 0.88, 0.60 to 0.91, 0.23 to 0.73 and 0.56 to 0.80, respectively. Cluster analysis of genotypes based on Jaccard’s similarity coefficient using DARwin generated a dendrogram which revealed 11 groups that followed clustering pattern more or less according their geographical origin as the Mantel’s test also indicated a good fit with r-value of 0.84. Internal transcribed spacer (ITS) sequence analysis revealed the distribution of Hippophae salicifolia in Sangla region of Kinnaur. Analysis of molecular variance (AMOVA) indicated sufficient genetic differentiation within populations and a low level of variation among populations. Clustering by DARWIN was supported by principal coordinate analysis (PCoA) and STRUCTURE. Some species specific diagnostic markers were also identified.

Key words: Hippophae, genetic diversity, simple sequence repeat (SSR), inter-simple sequence repeat (ISSR), Internal transcribed spacer (ITS), sequence analysis, analysis of molecular variance (AMOVA).