Abstract
Nothapodytes nimmoniana Graham; Syn. Mappia foetida (Icacinaceae) an endangered tree species of Western Ghats of India is an excellent source of quinoline alkaloids, camptothecin (CPT), used clinically as such or after derivation as anticancer agents for the treatment of solid tumors. Development of in vitro multiplication and conservation techniques are highly desirable, as there is no reported cultivation or conservation protocols of this endangered species. MS medium with different plant growth regulators (PGRs) were tried to optimize the protocol for in vitro multiplication of this species. Among the various explants tried, only the isolated seed embryos collected from Idukki district of Kerala, showed a positive response on MS with 0.91 μM thidiazuron (TDZ), 3% sucrose, 0.8% agar and pH 5.8 in Standard Culture Conditions (SCC). From regenerating callus, multiple shoots were regenerated and development of somatic embryos was observed on the same medium after 3 week of incubation. After one month of incubation in TDZ medium somatic embryos developed were small, inseparable and on an average of 70 per tube and 5 mm to 15 mm long. Shoot elongation of 2-19 mm was observed on MS basal medium after one month of incubation. One mg/L IBA was ideal for root initiation of the vitro plants. Some the cultures in (SCC) were shifted to low temperature (+10°C) for in vitro conservation. Under reduced temperature and light, in vitro plants grew well (without any subculturing) even after the six months of incubation. Cultures in MS basal medium showed shoot elongation of 5-10 mm compared to MS with TDZ (3-50 mm). The interesting results of growth response of the cultures in low temperature imply that it is possible to conserve the species under low temperature for a longer period without regular subculturing. Protocols for the establishment of in vitro gene bank for these species will be discussed.
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