Wang Zhihua, Song Jianhua, Zhang Yong, Yang Baoyu, Wang Yao, Chen Shiyun
State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, China.
Sheng Wu Gong Cheng Xue Bao. 2008 Oct;24(10):1707-13.
Studies have shown that transgenic plants expressing antiapoptotic genes from baculovirus and animals increase resistance to biotic and abiotic stress. However, the mechanism under these resistances is conjectural, or in some cases even controversy. In the present study, the p35 gene from baculovirus Autographa californica multicapsid nucleopolyhedrovirus (AcMNPV) was expressed in tobacco, and for the first time P35 protein was detected in transgenic plants by Western blotting. Inoculation of T1 transgenic tobacco leaves with tobacco mosaic virus (TMV) showed enhanced resistance, and DNA laddering was observed after TMV infection in control but not in transgenic plants. DAB staining showed that TMV infection did not affect peroxide induction of transgenic plants, Western blotting analysis of PR1 protein also showed no difference of control and transgenic plants. Inoculation of fungus (Sclerotinia sclerotiorum) using a detached leaf assay showed enhanced resistance of transgenic leave tissue. RT-PCR analysis demonstrated that p35 gene expression induced earlier expression of PR1 gene after S. sclerotiorum infection. Taken together, our results suggest that the mechanism under enhanced disease resistance by P35 protein is possibly related to the activation of PR-related proteins in addition to the inhibition of programmed cell death, depending on the pathogens challenged.
研究表明,表达杆状病毒和动物抗凋亡基因的转基因植物对生物和非生物胁迫的抗性增强。然而,这些抗性背后的机制只是推测,在某些情况下甚至存在争议。在本研究中,杆状病毒苜蓿银纹夜蛾多粒包埋核型多角体病毒(AcMNPV)的p35基因在烟草中表达,并且首次通过蛋白质免疫印迹法在转基因植物中检测到P35蛋白。用烟草花叶病毒(TMV)接种T1代转基因烟草叶片显示抗性增强,在对照植株中TMV感染后观察到DNA梯状条带,而转基因植株中未观察到。二氨基联苯胺(DAB)染色显示TMV感染不影响转基因植物中过氧化氢的诱导,PR1蛋白的蛋白质免疫印迹分析也显示对照植株和转基因植株之间没有差异。使用离体叶片试验接种真菌(核盘菌)显示转基因叶片组织的抗性增强。逆转录-聚合酶链反应(RT-PCR)分析表明,在核盘菌感染后,p35基因表达诱导PR1基因提前表达。综上所述,我们的结果表明,P35蛋白增强抗病性的机制可能除了抑制程序性细胞死亡外,还与病程相关蛋白的激活有关,这取决于所挑战的病原体。