Direct in vitro organogenesis from leaf and internode of Coccinia cordifolia (L.) Cogn.

The current study was carried out to elucidate a reproducible protocol to develop plants directly from leaf and internodes to facilitate the genetic transformation in Coccinia cordifolia (L.) Cogn., a medicinal plant of the cucurbitaceae family. In vitro grown leaf and internodes were used, which were collected for regenerated shoots from field-grown nodal segments that were sterilized by 0.1% HgCl2 treatment for 6 minutes. The nodal segment cultured on BAP, Kn and BAP, NAA combination where 1.5 mg/L BAP solitary supplement and augmented supplement with 0.1 mg/L NAA was most effective as 80% shoots regenerated with 4.0±0.37 and 2.7±0.45 shoots per culture, respectively. Collected leaf and internode responded 90% at 1.5 mg/L BAP + 0.1 mg/L NAA fortified full-strength MS medium. The highest number of shoots also regenerated in the same combination which were 8.1 ± 0.30 and 10.2±0.40, respectively and found internode as the best explant for direct organogenesis. For root induction, half strength of MS medium supplemented with 0.1 mg/L IBA was found most effective. The highest number of roots regenerated per shoot (6.8±0.10) and root length (2.8±0.20). The successful acclimatization of the in vitro (80%) grown plantlets proved the validity of the developed protocol of using biotechnological techniques for improving the plant

Propagation of Coccinia cordifolia (L.) Cogn. from shoot tip and nodal segment through micropropagation techniques

The present work was undertaken to develop a reproducible protocol for the micropropagation of an important medicinal plant of the Cucurbitaceae family, Coccinia cordifolia (L.) Cogn., by using shoot tips and nodal segments to overcome the impediment in seed settings and seed germination of conventional reproduction. To develop an efficient protocol, 0.1% HgCl2 treatment for 6 minutes was found effective for surface sterilization of field-grown explants to eliminate microbes and fungus and to get healthy tissues. Throughout the study, different concentrations of auxin, cytokine, and gibberellin were used either alone or in combination as supplemented in MS medium to find suitable conditions and suitable explant (nodal segment). BAP was the best cytokine source as 2.0 mg/l BAP produced 4.0±0.20 and 4.8±0.09 shoots per culture and gained 5.8±0.11 cm and 6.7±0.32 cm length with regeneration rate of 90 and 100 percent in shoot tips and node, respectively. The highest percentage (90%) of regeneration for axillary shoot proliferation was obtained in node explants at MS medium containing 2.0 mg/l BAP + 0.1 mg/l NAA. The highest number of shoots regenerated per culture was 3.0±0.41, with a length of 6.0±0.16 cm. GA3 was also found effective in producing longer shoots with the combination of BAP, but the average shoot number was reduced drastically. Although full-strength MS medium was found to be ideal for shoot regeneration and used in shoots proliferated experiments, half and full-strength of MS medium with auxins supplements of 0.5, 1.0, and 2.0 mg/l either of IBA, NAA, and IAA) were used for roots growth and half strength nutrients supplemented with 0.5 mg/l IBA was found most compelling. The highest number of roots regenerated per shoot was 3.1±0.30, and the average root length was 1.8±0.30 cm. Rooted plantlets were finally transplanted into small plastic pots containing sun sterilized sand, soil and humus (1: 2: 1) to adapt the plantlet in ex vitro environment, and acclimatized plantlets showed 95% survival rate in outdoor condition which proved the effectiveness of using biotechnology to improve plant’s growth rate and mass production