State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, Root Biology Center, South China Agricultural University, Guangzhou, 510642, China.
Plant Cell Rep. 2014 Nov;33(11):1921-32. doi: 10.1007/s00299-014-1669-5. Epub 2014 Aug 6.
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. 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.
在体外和体内毛状根转化系统都不能替代完整植物转化，用于分析大豆中根特异性和低磷诱导基因的启动子。一个有效的遗传转化系统对于植物启动子分析至关重要。农杆菌介导的转化是产生转基因毛状根或植物的最流行方法。在本研究中，首先，我们比较了两种不同的根癌农杆菌介导的毛状根转化方法，分别使用组成型 CaMV35S 启动子或根偏好基因 GmEXPB2 和 GmPAP21 的启动子，在大豆中，并发现体外毛状根转化的效率明显高于体内转化。我们比较了根癌农杆菌介导的毛状根转化和根癌农杆菌介导的整株植物转化系统。结果表明，低磷（P）诱导的 GmEXPB2 和 GmPAP21 启动子不能在体内和体外转基因毛状根中低磷胁迫下诱导 GUS 报告基因的表达增加。相反，在整株植物转化中，GmPAP21 启动子的 GUS 活性在低磷条件下显著高于高磷条件下。因此，在体外和体内毛状根转化系统都不能替代完整植物转化，用于分析大豆中根特异性和低磷诱导基因的启动子。
