Abstract
Simple and efficient protoplast culture procedure of eggplant was developed. Protoplasts were isolated from hypocotyls by the treatment with 0.067% Macerozyme R-10 and 0.33% Meicelase for 16 h at 25°C, and cultured in the medium with l/2 MS salts (200 mg/l NH4NO3), 1/2 MS vitamins, 1 mg/l 2, 4-D, 1 mg/l NAA, 1 mg/l kinetin, 1% sucrose and 0.3 to 0.5M mannitol. Auxin concentrations were reduced by the addition of the same medium without auxins at weekly intervals and finally to 0.1 mg/l of 2, 4-D and NAA. Mannitol concentration was also reduced gradually. Calluses larger than 3 mm were transferred to the MS medium with 0.2 mg/l IAA and 1 or 3 mg/l zeatin, which gave the highest shoot regeneration frequency. Elongated shoots were cultured in the MS Iiquid medium with 1 mg/l NAA for two days and transferred to the MS agar medium without hormones to regenerate roots. This culture procedure was applied to the protoplasts of various eggplant cultivars. Varietal differences were observed in the efflciency of colony formation and shoot regeneration. Regenerated plants were obtained efficiently from the protoplasts of 'Koushien', 'Khatkhatia long' and Solanum insanum.
