An efficient and rapid regeneration protocol was developed from nodal explants and callus cultures of Adenia hondala on Murashige and Skoog's (MS) medium supplemented with BAP and Kn. The highest number of shoots (8.25 ± 0.2) from nodal explants were obtained at an optimum combination of 0.5 mg 1−1 BAP + 0.5mg 1−1 Kn + 10% CW. The highest shoot elongation (8.25 ± 0.6 cms) was recorded at a concentration of 0.5 mg 1−1 BAP + 0.5mg 1−1 2 ip. Compact green callus was initiated from young internodal explants of MS medium supplemented with 3 mg 1−1 2, 4 – D + 1 mg 1−1 BAP. Organogenesis was achieved by transferring callus on MS medium supplemented with 1 mg L−1 BAP + 0.5 mg L−1 Kn. Highest number of shoots 14.25 ± 1.5 shoots/culture were initiated as clusters on the callus. In vitro regenerated shoots were then excised from shoot clumps and transferred to rooting medium containing half strength MS medium.
Somatic embryogenesis was obtained directly from leaf explants at a concentration of 2 mg 1−1 NAA + 0.5 mg 1−1 BAP + 200 mg 1−1 CH. 34 ± 0.2 embryos/culture were produced. Indirect somatic embryogenesis from friable calli was induced in 28 days at the optimum concentration of 1 mg 1−1 NAA + 0.5 mg 1−1 BAP + 0.5 mg 1−1 TDZ. The highest number of embryos 62.4 ± 2.5 per culture was produced at this concentration. After 15 days of culture globular embryoids became heart shaped and then torpedo shaped. The elongated embryoids showed distinct bipolar growth forming a shoot pole and root pole. The rooted plantlets were hardened on liquid half MS medium and subsequently in polycups containing sterile soilrite. Plantlets thus developed were successfully established and transferred to a green house. The plantlets showed survival rate of 90% in soil. Biochemical estimation of total protein, starch and soluble sugars when carried out in different stages of in vitro cultures indicated metabolic mobilization during organogenesis.
