ABSTRACT
Aims: The present studies were initiated to develop a cost effective protocol for micropropagation, as a mean for conservation of medicinal plant- Tylophora indica (Burm f.) Merill. The plant is threatened and needs immediate conservation, therefore, the study was undertaken with following objective: In vitro multiplication of Tylophora indica using nodal axillary bud proliferation and through organogenesis of callus. Study Design: For all experiments ten replicates were used per treatment and all the experiments were repeated three times. Data have been presented as Mean ± Standard deviation. Place and Duration of Study: Department of Plant Cell and Molecular Biology, Indian Institute of Advanced Research (IIAR), Gandhinagar, Gujarat, India. Methodology: For in vitro plant regeneration, micropropagation and organogenesis techniques were used. For micropropagation, surface sterilized nodal explants were inoculated on different shoot inducing media and further multiplication was obtained. Root containing shoots were transferred to pot containing pre autoclaved mixture of soil and soilrite. For organogenesis, surface sterilized leaf explants were inoculated on different types of callus inducing media. Results: All the nodal explants were sprouted at a very high frequency, i.e. 98% and sprouted buds elongated up to 8cm on three different media. Dissected explants grew further and average height of shoots reached 9.5cm±0.80cm within 30 days. Interestingly, root formation was observed on the same media; so that the best media was 0.4mg L-1 BA and 0.1mg L-1 Kn for both initiation as well as for multiplication. For organogenesis, the fragile callus was observed on media containing 2mg L-1 2,4-D and 0.1mg L-1 Kn. Green pigmented calli were transferred to MS media, where it regenerated in to shoots and roots, simultaneously Conclusion: The protocol of micropropagation through axillary bud proliferation described here is very simple, repetitive and cost effective, which can be easily utilized for commercial cultivation. On shoot multiplication media, root formation observed, thereby making the process is one step; which is very easy to follow.