陆地棉原生质体培养与植株再生技术研究

 分离培养陆地棉品种ZDM-3（Gossypium hirsutum L cv ZDM-3）胚性细胞悬浮系的原生质体，通过体细胞胚胎发生成功获得再生植株。试验结果表明，植物生长调节剂组合和碳源对原生质体培养具有重要的影响，添加2，4-D + KT + 1.5%（W/V）葡萄糖+ 1.5%（W/V）麦芽糖的KM8P培养基对于陆地棉原生质体培养的效果较好。激素组合0.46 μmol·L^-1 KT + 0.45 μmol·L^-1  2，4-D诱导的愈伤组织较松软，分化潜力高；胚性愈伤组织的增殖使用0.93 μmol·L^-1 KT + 2.46 μmol·L^-1 IBA的激素组合；1.2 μmol·L^-1 IBA + 1.39 μmol·L^-1 KT有利于体细胞胚胎发生，而MSB-5 + 0.67 μmol·L^-1 KT+2.69 μmol·L^-1 NAA的培养基适合体细胞胚胎萌发和植株再生。此外，愈伤组织诱导宜用葡萄糖作为碳源，而胚性愈伤组织增殖、保存和胚状体的萌发过程宜使用麦芽糖作碳源。

Study on the Plant Regeneration from Protoplasts of Upland Cotton (Gossypium hirsutum L.) Via Somatic Embryogenesis

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^-1 2，4-D，0.93 μmol·L^-1 KT，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 MSB5+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^-1 2，4-D could 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^-1 KT. The medium of MSB 5 + 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.