In vitro propagation of the elite species plant Pluchea lanceolata: Assessment of genetic stability by random amplified polymorphic DNA (RAPD) analysis

Abstract

An effective in vitro regeneration protocol was developed from nodal segment of Pluchea lanceolata (DC.) Oliver. & Hiern, a medicinally important plant used in ayurvedic system of medicine for curing diseases similar to rheumatoid arthritis. Nodal segments were cultured in MS medium supplemented with auxin and cytokinin and their combinations. The objective was to produce genetically identical plants, via multiple shoot induction from nodal segment. The culture medium consisted of Murashige and Skoog medium supplemented with one of 3 cytokinins [6-benzyladenine (BAP), Kinetin (Kn) and thidiazuron (TDZ)] at each of six different concentrations for shoot multiplication. The highest multiplication rate (24.57 shoots per explant) was obtained in the medium enriched with NAA. Shoots were successfully rooted in the half strength MS medium containing 0.1 µM NAA. In vitro produced plants were transferred to sterilized garden soil: compost (1:1) and then transferred to green house for hardening. Genetic stability of mother plant and the regenerants produced in vitro was assessed by random amplified polymorphic DNA (RAPD). In randomly selected plant material (mother plant) and its regenerants, 87 scorable bands were generated by four different primers, showing monomorphism with the mother plant. Thus, molecular analysis reveals that the micropropagation system described is a reliable method for propagation of P. lanceolata.

Keywords: Conservation, genetic fidelity, micropropagation, RAPD, TDZ.

African Journal of Biotechnology, Vol 13(25) 2500-2505