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采用花粉管通道介导法将含有bar基因的pPTN140质粒DNA导入8个黑龙江省大豆主栽品种(系),导入1384朵花,共获得1164粒种子,经2次Basta除草剂筛选后共获得3个品种的除草剂抗性植株8株。对其进行PCR以及PCR-Southern杂交检测均为阳性结果,Southern杂交结果显示,8个除草剂抗性后代植株整合数目不同,其中2株整合单拷贝基因,另外6株分别整合3~8个拷贝的bar基因。对8个抗性植株的后代进行了除草剂抗性分析,D1-D3代均出现除草剂抗性植株,表明bar基因可以在转基因植株后代中遗传,抗性遗传分析结果表明不同株系间以及株系内除草剂抗性缺乏规律性,分离比率不符合孟德尔遗传规律。说明大豆花粉管通道法转化基因存在多拷贝共抑制现象。目前获得除草剂抗性稳定的D3代株系82个。试验说明利用花粉管通道技术进行大豆转化是可行的。
The pPTN140 plasmid DNA containing bar gene was introduced into 8 main soybean cultivars (lines) in Heilongjiang Province by pollen tube channel-mediated method, and 1,384 seeds were introduced to obtain 1164 seeds. After 2 times Basta herbicide screening, a total of 3 A variety of herbicide-resistant plants 8. The results of Southern hybridization showed that the integrated number of eight herbicide resistant offspring was different, of which two integrated single copy gene and the other six integrated 3 ~ 8 copies respectively The bar gene. Herbicide resistance analysis was conducted on the progeny of 8 resistant plants. Herbicide-resistant plants appeared in D1-D3 generations, indicating that the bar gene can be inherited in the progenies of transgenic plants. The genetic analysis of resistance showed that among different strains and Herbicide resistance in plants lack of regularity, separation rate does not meet the Mendel’s genetic law. Soybean pollen tube channel method shows that there are multiple copies of the gene co-suppression phenomenon. At present, there are 82 D3 plants with stable resistance to herbicides. Tests show that the use of pollen tube channel technology for soybean conversion is feasible.