A comparison study of Agrobacterium-mediated transformation methods for root-specific promoter analysis in soybean
A comparison study of Agrobacterium-mediated transformation methods for root-specific promoter analysis in soybean
复制标题
大豆根特异性启动子分析农杆菌介导转化方法的比较研究
DOI：
10.1007/s00299-014-1669-5
复制
发表时间：
2014-11-01
影响因子：
6.2
通讯作者：
Liao, Hong
中科院分区：
文献类型：
--
作者：
Li, Caifeng;Zhang, Haiyan;Liao, Hong
高效的遗传转化系统是植物启动子分析的关键。农杆菌介导的转化是获得转基因毛状根或植株的最常用方法。本研究首先比较了发根农杆菌（Agrobacteriumrhizogenes，Agrobacteriumrhizogenes）介导的两种不同的大豆毛状根转化方法，即利用组成型CaMV35S和根偏好基因GmEXPB 2和GmPAP 21的启动子进行的转化，发现发根农杆菌的体外转化效率显著高于体内转化。我们比较了发根农杆菌介导的毛状根和根癌农杆菌介导的整株转化系统。结果表明，低磷诱导型启动子GmEXPB 2和GmPAP 21在低磷胁迫下不能诱导GUS报告基因在体内和体外转基因毛状根中的表达。相反，GmPAP 21启动子的GUS活性在低磷条件下显著高于高磷条件下。因此，在大豆根特异基因和低磷诱导基因的启动子分析中，无论是离体还是体内毛状根转化系统都不能代替整株转化。
An efficient genetic transformation system is crucial for promoter analysis in plants. Agrobacterium-mediated transformation is the most popular method to produce transgenic hairy roots or plants. In the present study, first, we compared the two different Agrobacterium rhizogenes-mediated hairy root transformation methods using either constitutive CaMV35S or the promoters of root-preferential genes, GmEXPB2 and GmPAP21, in soybean, and found the efficiency of in vitro hairy root transformation was significantly higher than that of in vivo transformation. We compared Agrobacterium rhizogenes-mediated hairy root and Agrobacterium tumefaciens-mediated whole plant transformation systems. The results showed that low-phosphorous (P) inducible GmEXPB2 and GmPAP21 promoters could not induce the increased expression of the GUS reporter gene under low P stress in both in vivo and in vitro transgenic hairy roots. Conversely, GUS activity of GmPAP21 promoter was significantly higher at low P than high P in whole plant transformation. Therefore, both in vitro and in vivo hairy root transformation systems could not replace whole plant transformation for promoter analysis of root-specific and low-P induced genes in soybean.