Analysis of microsatellite polymorphism in relation to geographical distribution and adaptation of seabuckthron in India

Abstract

Seabuckthorn (Hippophae rhamnoides L.) has long been recognized as a potential newlinesource of many bioactive compounds of immense medicinal and nutritional value. newlineConsidering the recent interest towards conservation and improvement of seabuckthorn as a future crop, efforts are being initiated towards assessment of genetic diversity newlineexisting in natural populations inhabiting diverse geographical areas. Compared to newlineother crop species, unfortunately, there is a scarcity of molecular tools particularly newlinemicrosatellite markers in seabuckthorn. Therefore, the present study was undertaken newlinewith the main objective of development of microsatellite markers following different newlineapproaches, and to evaluate these markers along with different morphological newlinecharacters in assessing genetic diversity in 93 collections of seabuckthorn representing newlinedifferent ecological sites in two geographical areas, Leh (Jammu and Kashmir) and newlineLahaul (Himachal Pradesh) in Indian Himalayas. Microsatellite markers were isolated newlineusing two approaches. The first approach involved screening of a microsatellite newlineenriched genomic library constructed following our novel and efficient methodology. newlineMicrosatellite positive sequences were fished out from a pool of restriction enzyme newlinedigested-linker ligated DNA fragments following hybridization with a biotin labeled newlineoligonucleotide probe and subsequent capture by streptavidin magnetic beads. A newlinesecondary PCR screening of recombinant clones, using linker and microsatellite probe newlineas primers, prior to sequencing of clones not only confirmed the presence of the newlinemicrosatellite repeat in the insert but also helped in inferring the location of the newlinemicrosatellite in the insert. newline newlineThe findings presented here are likely to find immense use in future breeding and molecular biology research projects in seabuckthorn aiming at its overall development as a crop.