High frequency plant regeneration from protoplasts in cotton via somatic embryogenesis
Wang, J.,Zhejiang Univ., Hangzhou (China). Dept. of Agronomy | Sun, Y.,Zhejiang Univ., Hangzhou (China). Dept. of Agronomy | Yan, S.,Zhejiang Univ., Hangzhou (China). Dept. of Agronomy | Daud, M.K.,Zhejiang Univ., Hangzhou (China). Dept. of Agronomy | Zhu, S.,Zhejiang Univ., Hangzhou (China). Dept. of Agronomy
A highly reproducible system for efficient plant regeneration from protoplast via somatic embryogenesis was developed in Gossypium hirsutum cultivar ZDM-3. Embryogenic callus, somatic embryos and suspension culture cells were used as explants. Callus-forming frequency (82.86%) was obtained in protoplast cultures from suspension culture cells in KM8P medium with 0.45 microM 2,4-dichlorophenoxyacetic acid (2,4-D), 0.93 microM kinetin (KIN), 1.5% glucose and 1.5% maltose. Protocolonies formed in two months with plating efficiency of 14%. However, the callus-forming efficiencies from other two explants were low. The calli from protoplast culture were transferred to somatic embryo induction medium and 12.7% of normal plantlets were obtained on medium containing 3% maltose or 1% of each sucrose + maltose + glucose, 2.46 microM indole-3-butyric acid (IBA) and 0.93 microM KIN. Over 100 plantlets were obtained from protoplasts derived from three explants. The regenerated plants were transferred to the soil and the highest survival rate (95%) was observed in transplanting via the new method.
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