PtWRKY14 基因转化烟草及对 TMV 的抗性影响研究

PtWRKY14 是从毛白杨中获得的WRKY 基因，为研究PtWRKY14 基因对植物抗病性的影响，构建了毛 白杨PtWRKY14 基因的植物表达载体pBI121-W14，通过农杆菌介导的叶盘转化法转化烟草，PCR 与Southern 杂交 进行转基因烟草鉴定。对获得的转基因烟草植株进行烟草花叶病毒(Tobacco mosaic virus，TMV）的接种试验，结果表 明PtWRKY14 基因的转化使烟草对TMV 的抗性增强。荧光定量PCR 检测结果表明，TMV 接种后，转基因烟草中 PtWRKY14 的表达量显著上调，同时转基因烟草中超氧化物歧化酶(Superoxide Dismutase，SOD）、过氧化物酶(Peroxidase，POD）、过氧化氢酶(Catalase，CAT）的表达量均高于对照组。因此，PtWRKY14 可能通过上调SOD、POD、 CAT 的表达从而增强植物抗病性。

Research on expression of PtWRKY14 in transgenic tobacco and resistance to TMV

PtWRKY14 is a WRKY gene from Populus tomentosa. The study was conducted to investigate the effects of PtWRKY14 on plant disease resistance. The gene PtWRKY14 was constructed into binary vector pBI121 and transformed into tobacco plants by Agrobacterium-mediated leaf disc transformation. Transgenic tobaccos were identified by PCR and Southern hybridization. The results of tobacco mosaic virus (TMV) infection test showed that transgenic tobacco possessed certain resistance to TMV infection. The expression of PtWRKY14, superoxide Dismutase (SOD), peroxidase (POD) and catalase (CAT) were analyzed by real-time PCR. The result showed PtWRKY14 was up-regulated expression by TMV treatment. While the expressions of SOD, POD and CAT in transgenic tobacco were more than in blank control tobacco. These results suggested that PtWRKY14 might be up-regulate expression of SOD, POD and CAT, leading to enhanced plant disease resistance.