Angiogenesis and Antioxidant Activity of in vitro and in vivo Tuber of Ceropegia pusilla Wight and Arn.

Background: Angiogenesis refers to the formation of new blood vessels. This biological process lays an important role in embryonic development. Ceropegia pusilla (Asclepiadaceae) used in folk medicine system to treat various health problems. Present work deals with the analysis of antioxidant and antiangiogenic properties of in vivo and in vitro tuber extracts of C. pusilla by Chicken Egg Chorioallantoic Membrane Assay (CAM) in vivo. Methods: C. pusilla tubers of both in vivo and in vitro were extracted with ethanol and methanol. In vivo anti angiogenesis effect was studied by Chicken Egg Chorioallantoic Membrane (CAM) assay. The antioxidant properties were analysed using 1, -diphenyl-2- picryl- hydrazyl free radical scavenging assay and reducing power activity. Result: Ethanolic extract of both in vivo and in vitro tuber showed higher angiogenic activities 58.3±13.6 and 43.2±12.2 respectively than the methanolic extract. The average number of vessels in in vivo and in vitro ethanolic extracts treated CAM was 6.48 and 5.43. In vivo and in vitro extracts exhibits comparable DPPH radical scavenging activity and better reducing power activity. Conclusion: C. pusilla tuber extracts of both in vivo and in vitro inhibited angiogenesis by blocking normal vascularisation in chick embryo and also having better DPPH and reducing power activity.

Microtuberization of Ceropegia pusilla Wight and Arn. an Endangered Medicinal Plant.

An efficient micropropagation (direct & indirect) and microtuberization protocol for an endemic and ethanomedicinal plant Ceropegia pusilla is described. The highest number of shoot initiation (3.50 0.34) and higher average shoot number in shoot subculture (15.40  1.65) were recorded on MS medium supplemented with BAP+NAA (2.5+0.5 mg/l). The best performance of callus induction and morphogenesis was found on MS medium supplemented with BAP and NAA from stem. The callus initiation was recorded on MS medium supplemented with BAP+NAA (0.5 + 1.0 mg/l) and the higher percentage of callus forming shoots from callus subculture is determinate to be 90-95  7.93. MS medium supplemented with various concentrations of cytokinins and auxins supported the induction of microtubers, in vitro flowering and roots. The microtubers and root formation occurred on the basal region of the shoots in MS medium containing BAP (1.5mg/l) and NAA (0.5mg/l). The higher average number of root length was (2.8  0.68cm) also observed in the same concentration. The plantlets regenerated in vitro with welldeveloped shoots, microtubers and roots were successfully established in pots containing hardening media and grown in a shade house with 81 3.16% survival rate.

Regeneration of Shoots from Callus of Ceropegia pusilla Wight and ARN.

This study was undertaken to evaluate the most suitable concentration of plant growth regulators and suitable explants for callus induction and subsequent organogenesis of an endangered medicinal plant Ceropegia pusilla. The best performance of callus induction and morphogenesis was found on MS medium supplemented with 6- benzylaminopurine (BA) and naphthalene acetic acid (NAA), from node and internode the maximum account of callus initiation was recorded on MS medium supplemented with BA+NAA (2.22μM + 5.37 μM) and the maximum % of callus induced on subculture is 5.47  0.68. Rooting was best achieved on MS medium augmented with IBA (2.46μM). The maximum number of roots (4.72  0.66 cm) and root length (4.8  0.49 cm) were recorded). Plantlets regenerated in vitro with well-developed shoot and roots were successfully acclimatized in pots containing a mixture of decomposed coir waste, perlite and compost 1: 1:1 ratio and grown in a shade house with 81  3.16 percent survival rates.