Study on Somatic Embryogenesis and Plant Regeneration of Theobroma Cacao L.
Plants were regenerated through somatic embryogenesis using cotyledon as explants. The optimized medium and condition for each stage were tested as follows ⑴ Primary calli medium PCG was the improved DKW with 3.0 mg/L of 24-D 1.0 mg/L of KT and 0.01 mg/L of TDZ explants were cultured on PCG medium in the dark at 28±2℃ for 20 days. ⑵ Calli propagation medium SCG was the improved DKW with 3.0 mg/L of 24-D 1.0 mg/L of KT the calli were cultured on SCG medium for 30 days in the dark. ⑶ The calli were subcultured on ED medium with 30 g/L of sucrose 1 g/L of glucose in the dark for 60 to 150 days and embryos at different developmental stages globular beart torpedo and mature were present on individual explants at the same time.⑷ The mature embryos were transferred to fresh PEC medium every 30 days until shoots with one or two leaves developed. The contents of PEC medium were the improved DKW with 20 g/L of glucose 10 g/L of sucrose. Shoot-producing embryos with 2 leaves were transferred onto RD medium that was the improved DKW supplemented with 1.5 mg/L of IBA and 0.5 mg/L of IAA. The cultures were maintained under light for 16 h/d. After 30 to 90 days regenerated plants were obtained.