EFFICIENT MICROPROPAGATION AND EVALUATION OF GENETIC FIDELITY OF IN VITRO RAISED PLANTS OF COMMIPHORA WIGHTII ARN. (BHANDARI) – A MEDICINALLY IMPORTANT RED-LISTED SPECIES OF ARID REGIONS

A refined and an efficient protocol for in vitro clonal propagation of Commiphora wightii, a red-listed desert plant of medicinal importance, has been developed from nodal segment of mature plant. Nodal segments from new branches having 6-7 nodes were excised after discarding the initial 5-6 cm terminal portion and were surface sterilized with 2.5% NaOCl (sodium hypochlorite), (v/v). MS medium [MURASHIGE & SKOOG, 1962] with different concentrations of BAP (6-benzylaminopurine) was used alone and in combinations with IAA (indole-3-acetic acid), NAA (α-naphthalene acetic acid), Kn (kinetin) and other additives for shoot induction. Best bud break response (84.5%) was obtained on MS medium supplemented with 8.88 µM BAP, 0.57 µM IAA and additives (50 mg l-1 ascorbic acid, 25 mg l-1 citric acid and 25 mg l-1 adenine sulphate) within 2 weeks of inoculation. The micro-shoots were subcultured and maintained for further elongation on the same medium for 4 weeks. Best shoot multiplication was obtained on same medium as used for shoot initiation. Best rooting was obtained when the shoots were initially given a 24 h pulse treatment in liquid MS medium supplemented with 4.92 µM IBA (indole-3-butyric acid) and 5.71 µM IAA under dark condition, followed by transfer to semi-solid half-strength hormone-free MS medium supplemented with 2% (w/v) sucrose and 0.5% (w/v) activated charcoal. High (86.7%) percent rooting was achieved after 4-5 weeks with 3-4 multiple adventitious roots of 5-6 cm length. These in vitro raised well rooted plantlets were acclimatized in a two step manner. During in vitro hardening step, the survival was 61.5% and during ex vitro hardening step it was 100%. Hardened plants (10-12 cm in height) were transferred to polythene-bags filled with mixture of soil and FYM in the ratio of 2:1 (v/v) and were kept in 75% agro-net shade for one month, where they gained height up to 60 cm. Five month old hardened plants were planted in open field condition for evaluation of these tissue cultured raised plants. There is cent percent survival of these field grown plants over period of two years with no visible morphological abnormalities. Genetic fidelity test was carried out for these in vitro raised plants by using RAPD primers (OPA and OPN). Uniform banding pattern was observed in all plants without any polymorphism.