IN VITRO REGENERATION OF THEVETIA PERUVIANA PERS.K.SCHUM, FAMILY APOCYNACEAE

Abstract

The objective of this study was to develop and optimize  in vitro protocol for regeneration of soma clonal variants of  T.peruviana  aimed at obtaining glycoside free or low glycoside plantlets with high oil and proteins content through tissue culture technique in order to develop new varieties which has novel traits compared to the existing landraces. T.peruviana seedlings germinated from mature fruits of orange flowering variety of T. peruviana  trees were used. Young but enlarged leaves next to the shoot tip were excised as source of explant. Callus initiation media consisted of MS salts, vitamins plus 8 g / l agar agar, 30 g  / l sucrose, 2.0 mg / l dichlorophenoxyacetic acid (2,4-D) supplemented with 0.1 mg / l of kinetin at pH of 5.75 at  22oC to 24oC and 12 hour darkness during the night and 12hrs lighting from fluorescent tubes during the day. Shoot regeneration medium consisted of MS media salts, vitamins, supplemented with 3 mg / l of 2 ip, 8 g / l agar agar, 30 g / l of sucrose, at pH of 5.75 were incubated at 250C-270C, with continuous lighting from florescence tubes in the growth chamber. Rooting was initiated by incubating single shoots into each media bottle containing 30 ml of MS media salts, vitamins, 1 g of glycine, 0.2 g of biotin, 3 mg/l of Indole-3- butyric acid (IBA), 8 g of agar agar and 30 g/l sucrose. It was noted that plantlets of T.peruviana regenerated after 12-24 months.