In vitro multiplication of pointed gourd (Trichosanthes dioica) through nodal explant culture, and testing the genetic fidelity of micropropagated plants using RAPD markers
Kumar, S.; Singh, H.; Pandey, V.; Singh, B. D.
Indian Journal of Biotechnology 15(4): 581-588
An efficient in vitro propagation system was developed for large-scale multiplication of a high yielding line (IIVRPG-102)of pointed gourd (Trichosanthes dioica Roxb.)utilizing nodal segments. A procedure for surface sterilization, involving a combination of treatments with cetrimide (1.0%) for 30 min, trimethoprim (0.05%) and dimethycarbendazim (0.2%) for 2 h, and finally with 0.1% HgCl2 for 3 min, was standardized yielding 87% contamination-free cultures. Plant growth regulator (PGR)-free MS medium was adequate for inducing bud break in up to 81.5% of the explants. Addition of PGRs like NAA, and thiadizuron had a small but significant effect on the frequency of responding explants. The highest number of shoots per explant (7.6) was obtained on MS medium supplemented with 1.0 mg L-1 BA. Root induction was achieved with NAA or IBA treatment. The best rooting responses were obtained on a medium containing 0.1 mg L-1(89.0%) or 0.2 mg L-1 IBA (87.0% rooted shoots). The micropropagated shoots with well-developed roots were acclimatized and thereafter successfully transplanted to a glasshouse with 90.6% survival rate. PCR analysis with 25 random decamer primers yielded monomorphic products in the micropropagated plants identical to those of the mother clone, confirming the genetic uniformity of the regenerated plants. This protocol could be applied to produce a large number of propagules of high-yielding selections to fulfill commercial demand.