利用双T-DNA载体系统培育无选择标记转基因大豆

GENERATING MARKER-FREE TRANSGENIC SOYBEAN PLANTS BY AGROBACTERIUM-MEDIATED TRANSFORMATION WITH DOUBLE T-DNA BINARY VECTOR

利用PCR和Southern杂交检测了161株转△6-脂肪酸脱氢酶基因(D6D)的T1代植株,初步筛选出只含有功能基因(D6D)而不含有筛选标记基因(bar)的转基因大豆植株9株,为获得富含γ-亚麻酸但不含选择标记基因的转基因大豆奠定了基础,并为转基因大豆的安全性种植提供了保障。同时对无标记基因的转基因植株进行了叶片涂抹除草剂验证,探讨了叶片涂抹除草剂结合目的基因PCR检测筛选无选择标记转基因植株的可行性。

Transgenic crops have developed in commercialized scale in some countries, but their biosafety have attracted public concerns in the world at the same time. Therefore, it is becoming more and more important to eliminate the selective genes because of their negative functions, such as antibiotics resistance and herbicide resistance. 161 T1 transgenic soybean of pudou 8008 containing △^6- fatty acid desaturase gene and bar selection gene, which were obtained by biolistic particle co-transformation, were analyzed by PCR and southern blot in this study, and 9 plants only with the target gene were identiffed finally. Moreover, leaf painting of herbicide was applied to confirm the marker free transgenic plants. It could be more efficient to combine leaf painting and PCR, Southern blot to select bar free transgenic soybean.