雄性不育基因对棉花的遗传转化

Cotton Genetic Transformation of Barnase Male Sterility Gene

利用TA29、A6、A9三启动子功能区与barnase基因融合构建的不育基因以农杆菌介导法对棉花下胚轴进行了遗传转化,通过胚状体途径获得了转基因再生植株。利用150 mg.L-1高浓度卡那霉素(Km)对转化初期筛选出的再生苗进行再次筛选,提高了转化株的选出率。通过PCR检测和Southern dot blot分析从转基因胚状体再生植株中获得了带有barnase不育基因的120株转基因植株。进行转基因植株生物学性状检测和观察表明,所获得的转基因植株对溴苯腈表现出了明显的抗性,并从不育基因转化植株中筛选出了具有明显不育特征的雄性不育株。

Cotton male sterility mutations naturally have been proved to be useful for producing hybrids. Crosses between cotton lines often result in progeny with higher hybrid vigor. However, the hybrid production has been limited because there is no ideal fertility restoration variety. The establishment of genetic engineering strategy of male sterility and fertility restoration varietyies has facilitated the development of breeding system for hybrid cotton. To obtain genetic modified male sterile cotton, the chimaeric male sterile gene which was constructed via placing the ribonuclease barnase under the control of anther-specific promoter was used to transform hypocotyledonary axis of cotton via Agribectrum tumefaciens. Embryoids were generated from transformed calli and then developed to seedling. 150 mg · L^-1 Kanamycin was used for selecting positive transgenic seedling from the primary one from embryoids and many negative transgenic seedlings were screened out. 120 lines of transgenic male sterility were identified via PCR and Southern dot blotting. 69 lines of the identified transgenic plants were conferred a resistance to bromoxynil and male-sterile traits, however, only 6 lines showed significantly male-sterile traits.