Organogenesis and plant regeneration in Taxus wallichiana (Zucc.)

We describe an efficient process for regeneration of Taxus wallichiana plants via shoot organogenesis from callus cultures derived from zygotic embryos. Zygotic embryos cultured on half strength Lloyd and McCown's basal medium supplemented with SH vitamin (1/2 WPMSH), 0.5 mg l-1 6-benzyladenine (BA) in combination with 1.0-2.0 mg l-1 2,4-dichlorophenoxyacetic acid (2,4-D) or α-Napthaleneacetic acid (NAA) produced two morphologically distinct types of calli-compact, green callus (CG) and compact, yellow (CY) callus after 4 weeks of culture. Optimum frequency (63%) of adventitious shoot bud induction was achieved in CG callus (3.0±0.67 shoot buds per gram of CG callus) when cultured on 1/2 WPMSH basal medium supplemented with 2.5 mg l-1 BA after 4 weeks. The inclusion of 1% activated charcoal (AC) to 1/2 WPMSH basal medium (shoot elongation medium) led to maximum shoot elongation (2.15 cms). Microshoots rooted in high frequency (40%) in MS basal medium in which the concentration of nitrates was reduced to one-fifth the normal concentration after 4 months of culture.