Reimann-Philipp U, Beachy R N
Department of Biology, Washington University, St. Louis, MO 63130.
Mol Plant Microbe Interact. 1993 May-Jun;6(3):323-30. doi: 10.1094/mpmi-6-323.
Coat protein-mediated resistance (CP-MR) was studied in transgenic Nicotiana tabacum 'Xanthi nn' and 'Xanthi NN' that express chimeric tobacco mosaic virus (TMV) coat protein (CP) gene constructs using two different tissue-specific promoters. The Phaseolus vulgaris pal2 promoter leads to gene expression in the upper leaf epidermis and the xylem, while the rolC promoter from Agrobacterium rhizogenes leads to gene expression in pholem and leaf hair tip cells. Tissue-specific gene expression was verified using the gusA(uidA) reporter gene, while accumulation of TMV CP was verified by Western blot analysis. Transgenic Xanthi nn plants harboring the pal2-CP gene construct were partially resistant to TMV infection. On Xanthi NN plants that expressed the pal2-CP gene construct, fewer necrotic lesions were formed after TMV inoculation compared to nontransformed control plants. The level of resistance, however, was substantially less than in plant lines that expressed TMV CP from the cauliflower mosaic virus 35S promoter. By contrast, expression of the rolC-CP construct did not confer resistance in either Xanthi nn or Xanthi NN. The results provide further evidence that CP-MR to systemic TMV infection in tobacco is probably due to inhibition of infection rather than to effects on long-distance spread through the phloem.
利用两个不同的组织特异性启动子,在表达嵌合烟草花叶病毒(TMV)外壳蛋白(CP)基因构建体的转基因烟草“Xanthi nn”和“Xanthi NN”中研究了外壳蛋白介导的抗性(CP-MR)。菜豆pal2启动子导致基因在上部叶片表皮和木质部中表达,而发根农杆菌的rolC启动子导致基因在韧皮部和叶尖毛细胞中表达。使用gusA(uidA)报告基因验证组织特异性基因表达,同时通过蛋白质免疫印迹分析验证TMV CP的积累。携带pal2-CP基因构建体的转基因Xanthi nn植株对TMV感染具有部分抗性。在表达pal2-CP基因构建体的Xanthi NN植株上,与未转化的对照植株相比,TMV接种后形成的坏死病斑较少。然而,抗性水平明显低于从花椰菜花叶病毒35S启动子表达TMV CP的株系。相比之下,rolC-CP构建体的表达在Xanthi nn或Xanthi NN中均未赋予抗性。结果提供了进一步的证据,表明烟草中对TMV系统感染的CP-MR可能是由于感染受到抑制,而不是对通过韧皮部长距离传播的影响。