棉 花 学 报     Cotton Science   2008，20(6)：403－407

Study on the Plant Regeneration from Protoplasts of Upland Cotton ( Gossypium hirsutum  L.) Via Somatic Embryogenesis
WANG Jing-er，SUN Yu-qiang，YAN Shu-feng，DAUD Muhammad-khan，SHEN Xiao-jia，ZHU Shui-jin^*
(College of Agriculture and Biotechnology，Zhejiang University，Hangzhou 310029，Zhejiang，China)

Abstract：Efficient plant regeneration from protoplast isolated from suspension culture cells via somatic embryogenesis was developed in upland cotton ( Gossypium hirsutum L.) cultivar ZDM-3. Microcalli were formed after being cultured in KM8P medium with 0.45 μmol·L^-12，4-D，0.93 μmol·L^-1KT，1.5%(W/V) glucose，1.5%(W/V) maltose and 9.0%(W/V) mannitol for about two months. Calli with 2 mm in diameter were transferred to solid proliferated medium. A great deal of somaticembryos and regenerated plants were occurred in the medium of MSB₅+2.46 μmol·L^-1 IBA+0.93 μmol·L^-1 KT+3%(W/V)glucose + 0.25%(W/V) phytagel. The regenerated plants were grafted to transplant ， survival percentage was above 95%. The results showed that A cotton protoplast culture system for ZDM-3 was highly improved by the three key steps namely，high proto-callus forming efficiency，high conversion rate of somatic embryo to plantlets and high survival probability of transplanting. The results showed further that there were great effects of the combinations of the plant growth regulators and sugar kinds on the upland cotton protoplast culture，the KM8P medium supplement with 2，4-D + KT + 1.5%（W/V）glucose +1.5%（W/V）maltose had the best result of the protoplast culture. The hormone combination of 0.46 μmol·L^-1 KT + 0.45 μmol·L^-12，4-Dcould produce the best quality callus with watery texture，which had the high capability of plant regeneration. The embryogenic callus proliferated was better in the medium with 0.93 μmol·L^-1 KT and 2.46 μmol·L^-1，while somatic embryo initiated when IBA concentration in the medium was decreased to 1.2 μmol·L^-1 IBA and KT was increased up to 1.39 μmol·L^-1KT. The medium of MSB₅+0.67 μmol·L^-1 KT+2.69 μmol·L^-1 NAA was suitable for the germination of somatic embryos and plant regeneration. In addition，the best sugar for callus inducing was glucose，while the maltose was even better for the proliferation of embryogenic callus and germination of somatic embryo.
Key words：Gossypium hirsutum L.；protoplast culture；somatic embryogenesis；plant regeneration
Abbreviation：2，4-D:2，4-Dichlorophenoxyacetic acid；NAA:α-Naphthaleneacetic acid；IBA: Indole-3-Butyric Acid ；KT: Kinetin；ZT: Zeatin      [Full Text, 2908KB]