农杆菌介导磷高效转录因子OsPTF1转化大豆研究

Transformation of OsPTF1 into Soybean by Agrobacterium-mediated Method

本研究将水稻耐低磷基因OsPTF1基因构建植物高效表达载体pTF101.1-35S-OsPTF1-NOS,通过农杆菌介导法将OsPTF1基因转入黄淮海地区优良品种商豆1099子叶节中,成功获得转基因植株。大豆属于难转化作物,在本研究中,对大豆转基因中大豆萌发指标和侵染方式进行优化选择。结果表明:大豆转基因萌发时间以48 h为宜;大豆子叶节受体比大豆胚尖受体更容易转化。经过PCR分子初步检测,有3株转化植株整合有OsPTF1外源基因,表明OsPTF1基因已初步整合到大豆基因组中。

In this study, a binary vector pTFlOl.l-35S-OsPTF1-NOS was constructed from the rice OsPTF1 gene, a transcription factor involved in tolerance to phosphate-starvation in rice. A elite commercial bean cultivar in Huang-Huai-Hai region, Shangdou 1099, was successfully transformed with the vector via Agrobacterium-mediated transformation method. The transformation conditions and parameters was assessed with the consideration of germinating time and infection pattern. The results showed that the optimal germinating time is 48 hours, and that cotyledonary node is the better recipient of transformation than embryonic tips. The gene of OsPTF1 was amplified by PCR from 3 transgenic lines, indicating that the gene had integrated into the bean genome.