An efficient protocol for clonal regeneration and excised root culture with enhanced alkaloid content in Thalictrum foliolosum DC.— an endemic and important medicinal plant of temperate Himalayan region

Abstract

Thalictrum foliolosum DC. is an endemic herbaceous plant known for its various medicinal properties. Due to the presence of valuable alkaloids they are uprooted leading to a threat to their numbers and existence. In this study an efficient protocol has been developed for plant regeneration using hypocotyl explant. Hypocotyl explants formed multiple shoots via direct shoot organogenesis on Murashige and Skoog’s (MS) medium containing 0.25–2.0 mg l−1 BAP with NAA (0.1 mg l−1). Maximum shoot formation in hypocotyl regenerated shoots was achieved in BAP 1 mg l−1and NAA 0.1 mg l-1 with 0.3% activated charcoal and ascorbic acid 20 mg l−1. The regenerated shoots formed roots on MS medium (half strength) containing IBA (0.2 mg l−1). The rooted plantlets were established in the glasshouse after acclimatization and gave 72% survival. Genetic stability assessment of in vitro regenerated plants was done using simple sequence repeat (SSR) markers and no difference was observed when compared to the mother plants. Also, flow cytometric analysis confirmed that their ploidy level was similar to the mother plant. Additionally, establishment of excised root culture was also done and quantification by HPLC revealed that excised cultured roots were accumulated higher benzylisoquinoline alkaloids (BIAs).