Efficient plant regeneration via indirect organogenesis and genetic stability assessment by molecular markers in an endangered medicinal plant Operculina turpethum (L.) Silva Manso

Abstract

Operculina turpethum (L.) Silva Manso is reported to be effective against several diseases including tumor, jaundice, gastrointestinal disease etc. The plant is facing extinction due to overexploitation and inefficient conservation strategies. In this work, an effective approach for micropropagation of O. turpethum by indirect organogenesis from stem explant is devised. Induction of callus culture was carried out on Murashige and Skoog medium enriched with benzyl adenine (3mg/L). Subsequently, the callus cultured on MS medium with 1mg/L BA + 1mg/L kinetin generated a maximum of 3.20 ± 0.7 number of shoots/0.5gm callus within 35 days of incubation. On MS medium with Indole-3-acetic acid (IAA)(0.2mg/L), 98.4% of the shoots were successfully rooted. The in vitro regenerated plantlets were acclimatised and shifted to outdoor condition with a success rate of 95%. The genetic stability of micro propagated plantlets was assessed using PCR-based molecular markers, random amplified polymorphic DNA (RAPD) and inter simple sequence repeat (ISSR) which yielded a total of 27 and 31 scorable bands respectively. The monomorphic banding pattern of in vitro propagated plants confirms their genetic homogeneity. This work is report on in vitro plant regeneration through indirect organogenesis and evaluation of genetic fidelity of micropropagated plantlets of O. turpethum.