Abstract
Owing to the high demand, Swertia chirayita populations in the wild are being depleted beyond its regeneration capacity. S. chirayita is one of the most valuable medicinal plants of Nepal in trade. Present Molecular investigation was undertaken to understand the level of genetic diversity in five S. chirayita populations of Nepal using Polymerase Chain Reaction (PCR)-based Random amplified polymorphic DNA (RAPD) technique. Thirty four accessions of S. chirayita along with six outlier accessions were analyzed using 26 arbitrary primers. Of the total 285 amplified bands scored for S. chirayita, 263 bands (92.28%) were polymorphic. Two major clusters were revealed in the phenogram generated from cluster analysis using NTSYS-PC software (version 2.21i) for the geographic populations under study. Principal Coordinate Analysis further substantiated the results of the phenograms. Swertia chirayita populations from Sankhuwasabha and Terathum were found to be genetically closest (68%, similar) whilst Nagarjun and Terathum were found to be most distant (33%, similar).The high genetic polymorphism reflected in S. chirayita populations indicates the good survival potentiality and adaptability in changing environmental scenario. The results thus produced might be helpful to plant breeders for elite cultivar development. The RAPD-PCR technique is found to be the rapid and effective tool for genetic diversity assessment in S. chirayita populations and generated insights for the formulation of conservation strategy of this vulnerable species together with its phytochemical distinctiveness.
Highlights
Nepal is very rich in medicinal plants and their associated traditional knowledge
The optimized Random amplified polymorphic DNA (RAPD)-Polymerase Chain Reaction (PCR) reaction parameters consisted of DNA (25 ng), MgCl2 (3.0 mM), 10X Taq polymerase reaction buffer [2.5 μl; 100 mM Tris-HCl, 500 mM KCl, 0.8% (v/v) Nonidet P40], Taq DNA polymerase (1U, MBI FERMENTAS company), dNTPs (0.2 mM each) and primer (0.4 μM) in 25 μl PCR reaction volume
Program consisted of an initial denaturation step at 95 ̊C for 2 min followed by 45 cycles of 95 ̊C for 20 s, 37 ̊C for 60 s and 72 ̊C for 60 s and final extension of 72 ̊C for 10 min
Summary
Nepal is very rich in medicinal plants and their associated traditional knowledge. Over 1950 medicinal plants have so far been reported [1]. Medicinal plants of Nepal are increasingly depleting and threats to these high value resources are from land fragmentation, degradation, overexploitation as well as prevailing climate change. S. chirayita has been listed as one of the top priority medicinal plants of Nepal as well as of Asia [2]. Member genus of the family Gentianaceae is widely distributed globally and is represented by over 135 species distributed in temperate areas of Asia, Africa, Europe, North America and Madagascar [3,4]. Owing to diverse physiographic zones, climatic contrasts and alti-
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