VARIOUS METHODS FOR SEEDLINGS PRODUCTION OF SMILAX OLDHAMI MIQ. BY TISSUE CULTURE

Axillary bud was easily induced from nodal segment placed on a modified half strength Murashige and Skoog (MS) medium without hormones. The rooting from the shoot was promoted by 0.5 mg/l NAA in the medium. Multiple buds were formed directly on the cut end of petiole, when the explants were cultured on ½ MS medium supplemented with 1 mg/l BA. Somatic embryogenesis was observed on the surface of leaf segment, which was cultured on ½ MS medium with 1 mg/l BA for two weeks, successively subcultured on the medium with 2 mg/l 2,4-D, then transferred to the hormone free medium. The development of shoot bud and root from the somatic embryos was promoted by 1 mg/l kinetin and 0.02 mg/l NAA in the medium. It was observed with shoot internode explants that plantlets regenerated through embryogenesis via callus and the direct formation of adventitious shoot. For these organogenetic events, the combination of BA(0.1 mg/l) and NAA(0.1 mg/l) in the medium was favorable. Similar hormonal combination and concentration gave the best results for the formation of callus and embryogenic callus on the root segment. Shoots formed from petiole, shoot internode and root segment rooted easily on ½ MS medium without hormones.

After acclimatization, these regenerated plants have grown normally in the soils for more than two years.