IN VITRO SHOOT REGENERATION AND MICROCORM DEVELOPMENT IN CROCUS VERNUS (L.) HILL

An efficient method has been developed for In vitro regeneration of shoot and microcorm from corm explants of Crocus vernus. Corms were cut into 0.5-1.0 cm long segments and cultured on the SH medium supplemented with 0.5, 1.0, 2.0, or 4.0 mg L-1 2-isopentyl adenine (2-iP), N-6-benzyladenine (BA), and N-6-furfuryladenine (kinetin, Kin) alone or combination with 0.5 or 1.0 mg L-1 alpha-naphthalene acetic acid (NAA) for shoot regeneration. Of the three cytokinins tested, BA was found to be the most effective cytokinin for shoot formation. The number of shoots induced per explant was more when BA was combined with 0.5 mg L-1 NAA than with 1.0 mg L-1 NAA. The greatest percentage of shoot induction (97.2) with the mean number of 11.8 shoots per explant was obtained when the SH medium was supplemented with 2.0 mg L-1 BA and 0.5 mg L-1 NAA. The frequency of microcorm induction was significantly affected by the concentrations of sucrose. The greatest number of 6.1 microcorms per explant was obtained when the SH medium was supplemented with 2.0 mg L-1 BA, 0.5 mg L-1 NAA and 6.0% sucrose. The microcorms formed In vitro developed daughter corms when they were cultured on this medium. Microcorms were separated from the culture and planted out in acclimatization boxes containing a commercial medium. About 85% of corms developed shoot and root after 30 days. This protocol could be utilized for genetic transformation and mass clonal propagation of C. vernus.