利用农杆菌介导法转化泗棉3号的研究

泗棉3号农艺性状优良，是20世纪90年代长江流域棉区的主栽品种，利用基因工程导入外源基因进行直接改良，可以迅速获得新的品种或育种材料。以陆地棉泗棉3号的下胚轴为外植体，建立了高效的转化体系，得到了大量的转基因植株。泗棉3号的出愈率和分化率显著高于模式品种Coker 312，同时它出现分化中心的主要形态不同于Coker 312，其胚性愈伤组织主要来源于两类初生愈伤组织。对泗棉3号的胚性愈伤组织进行GUS检测，以及对再生植株叶片进行PCR检测后，阳性植株嫁接于温室。在温室用2 063.98 μmol L^-1的卡那霉素点涂叶片检测nptⅡ基因的表达和取叶片进行gus基因的组织化学检测，同时通过Southern blot分析检测，证明目的基因成功地整合到泗棉3号基因组中。

Agrobacterium-Mediated Transformation of Upland Cotton Simian 3

Simian 3 is one of major cultivars at the cotton-growing district in Yangtze River Valley in the 1990s. New breeding materials or varieties can be rapidly developed through direct transformation of exogenous genes. In this study, we generated transgenic plants from hypocotyl segments of Simian 3 by Agrobacterium-mediated transformation. The rates of callus induction and differentiation were higher in Simian 3 than those in Coker 312. And the major callus morphological characters of Simian 3 from which differentiation center could be formed differed from Coker 312. Histochemical GUS (β-glucuronidase) assay and PCR for nptⅡ (neomycin phosphotransferase) gene were carried out with the transgenic embryogenic callus and leaves of transgenic plantlets. The plantlets proved by histochemical GUS assay and PCR for nptⅡ were grafted and then cultivated in greenhouse. The application of kanamycin at the level of 2 063.98 μmol L^-1 and Southern blot also proved that the target gene was successfully integrated into the genome of Simian 3.