农杆菌介导的bar基因和Bt基因的遗传转化研究  

Studies on Genetic Transformation of Agrobacterium-mediated Bar Gene and Bt Gene

作　　者：孟灵真[1] 陈全家[1] 杨婷[1] 阿依夏木.古丽 王希东[1] 曲延英[1] 

机构地区：[1]新疆农业大学农学院/新疆农业大学农业生物技术重点实验室,乌鲁木齐830052

出　　处：《新疆农业科学》2013年第12期2189-2196,共8页Xinjiang Agricultural Sciences

基　　金：国家转基因重大专项项目(2011ZX0800500500-008)

摘　　要：【目的】探讨不同基因对不同棉花品种的遗传转化差异。【方法】以新海14号、新海16号、新海24号和新海30号四个不同基因型海岛棉胚性愈伤组织为受体材料,通过农杆菌介导法分别将pBI121-bar/Bt和pBI121-Bt转化至四个品种中,对遗传转化体系中的PPT浓度和Cef浓度进行筛选,观察比较不同浓度下不同基因型胚性愈伤的生长状况,并对单价表达载体和双价表达载体转化不同基因型胚性愈伤的生长状态进行比较。【结果】转化体系的筛选确定双价表达载体的PPT筛选浓度为3.0 mg/L,Cef的筛选浓度为800 mg/L;转化单价表达载体LBA4404/pBI121-Bt的Cef的筛选浓度为600 mg/L。转化单价表达载体后,新海14号的出愈率最高,新海30号的出愈率最低;转化双价表达载体后,新海14号的出愈率最高,新海24号的出愈率最低。【结论】不同基因型对PPT的敏感性有差异;转化不同基因后,胚性愈伤对抗生素的敏感度有所不同;转化相同基因后,不同品种的胚性愈伤生长存在差异。【Objective】To study the difference of genetic transformation of difference genes that bring to different cotton strains.【Method】Embryogenic callus of four different genotypes of cotton XH 14,XH 16,XH 24 and XH 30 were taken as the receptor materials, and through agrobacterium- mediated method, LBA4404 / pBI121- bar / Bt and LBA4404 / pBI121- Bt were transformed into the four species. PPT and Cef concentration in genetic transformation system were screened to observe their growth status of different genotypes and concentration of embryogenic callus,and compare the growth status with the single and bivalent expression vector.【Result】The results showed that the optimal concentrations of PPT was 3. 0 mg / l and Cef was 800 mg / l after transforming bivalent expression vector of LBA4404 / pBI121- bar / Bt. But the optimal concentrations of Cef was 600 mg / l after transforming single expression vector of LBA4404 / pBI121- Bt. The callus rate of XH 14 was the highest and the lowest one was XH 30 after transforming single expression vector. Identically,the callus rate of XH 14 was the highest and the lowest one wasXH 24 after transforming bivalent expression vector.【Conclusion】It indicated that the sensitivity of different genotypes to PPT was different,when transforming different genes,the sensitivity of embryogenic callus to different antibiotics was different,but when transforming the same genes,the growth of the embryogenic callus of the different strains was different.

关 键 词：PPT Cef基因型 胚性愈伤组织 遗传转化 

分 类 号：S562[农业科学—作物学] S503.53