Plant Regeneration from Cotyledon Explants of Bittergourd as influenced by Adenine Sulphate and D-Biotin

The first experiment was done to select the most suitable explant to establish a culture for shoot regeneration. Four different explants (cotyledon, cotyledon node, leaf and hypocotyl) excised from in vitro grown seedlings were cultured on MS medium containing 1.0 mg l-1 BAP and 0.2 mg l-1 NAA. Cotyledon was the most responsive explant which exhibited quick callus initiation. The second experiment was conducted to determine the effect of cotyledon explants on different MS media containing 1.0 mg l-1 BAP, 0.2 mg l-1 NAA and also various concentrations (0, 20, 40 and 60 mg l-1) of adenine sulphate (AS). The best medium for callus formation was MS medium with 1.0 mg l-1 BAP, 0.2 mg l-1 NAA and 60 mg l-1 AS. Callus derived from the particular medium also showed the best positive response for shoot regeneration after subculturing on MS medium with 2.0 mg l-1 BAP and 0.2 mg l-1 NAA. In the third experiment, D-biotin added into the composition of the culture medium had a beneficial effect on the callogenesis in association with BAP and NAA. Regenerated shoots were multiplied on MS medium with 3.0 mg l-1 BAP. Eventually, in vitro rooting was achieved on MS medium included with 1.0 mg l-1 IBA.