Congruence of inter simple sequence repeats (ISSR) and random amplification of polymorphic deoxyribonucleic acid (RAPD) markers in genetic characterization of Artemisia annua in the Trans-Himalayan region

Abstract

Artemisia annua is an important medicinal plant valued all over the world. Genetic characterization of 20 genotypes of A. annua collected from two valleys viz. Nubra (9,600 ft) and Leh (11,500 ft) of the Trans-Himalayan (Ladakh, India) region were analyzed using 37 polymerase chain reaction (PCR) markers (20 random amplification of polymorphic deoxyribonucleic acid. (RAPDs) and 17 inter simple sequence repeats) (ISSRs). RAPD analysis yielded 124 polymorphic fragments (96.9%), with an average of 6.2 polymorphic fragments per primer. ISSR analysis produced 85 bands, of which 78 were polymorphic (86.1%), with an average of 4.58 polymorphic fragments per primer. The primers based on (CT) n produced maximum number of bands (nine) while, (AT) n and many other motifs gave no amplification. The genetic diversity was high among the genotypes (Nei’s genetic diversity = 0.336 and Shannon’s information index = 0.495) as measured by combination of both RAPD and ISSR markers. The mean coefficient of gene differentiation (Gst) was 0.145, indicating 85.5% of the genetic diversity resided within the genotypes. RAPD markers were found more efficient with respect to polymorphism detection, as they detected 96.9% in comparison to 86.1% for ISSR markers. It was found that the genetic diversity among genotypes from Nubra valley was narrow than that of Leh valley, suggesting the importance and feasibility of introducing elite genotypes from different origins for Artemisia germplasm conservation and breeding programs. Key words: Artemisia annua, Ladakh, genetic diversity, random amplification of polymorphic deoxyribonucleic acid (RAPD), inter simple sequence repeats (ISSR), analysis of molecular variance (AMOVA).