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PCR和 Southern blotting检测表明 ,来自大肠杆菌的 mtl D基因已通过农杆菌介导整合进水稻基因组。mtl D基因在 T1代出现分离 ,T2 代出现纯系。在 0 .75% Na Cl胁迫下 ,7个转基因 T3 代株系都能检测到 mtl D酶活性 ,与对照相比细胞膜的相对电导率和大分子渗漏值明显降低。部分转基因株系能在 1 .0 % Na Cl浓度下正常生长 ,而对照在 0 .5% Na Cl浓度下已不能存活。通过有性杂交途径实现了 mtl D和 gut D两个基因的聚合 ,部分杂交后代株系能在 1 .2 5% Na Cl胁迫下正常生长结实。
PCR and Southern blotting showed that the mtl D gene from Escherichia coli has been integrated into rice genome by Agrobacterium tumefaciens. The mtl D gene was segregated in the T1 generation, and the pure line appeared in the T2 generation. At 0 .75% NaCl stress, mtl D enzyme activity was detected in all of the 7 transgenic T3 lines. Compared with the control, the relative conductivity and the macromolecular leakage of the membrane were significantly decreased. Some of the transgenic lines grew normally at 1.0% NaCl concentration, whereas the control did not survive at 0.5% NaCl concentration. The hybridization of mtl D and gut D genes was achieved by sexual crossing, and some hybrid progeny lines could grow normally under the stress of 1.25% NaCl.