论文部分内容阅读
来源于昆虫病毒和动物的抗细胞凋亡基因能够诱导植物对生物或者非生物胁迫产生抗性,但其抗性机理有不同甚至相反的报道。本研究将来源于苜蓿银纹夜蛾核多角体病毒的p35基因转化烟草,T1代转化烟草Western blotting检测P35蛋白的表达,转化烟草接种烟草花叶病毒(Tobacco mosaic virus,TMV)抗病效果增强。进一步的抗病机理研究表明,转化和野生型烟草感染TMV后诱导过氧化氢积累无明显区别,野生型烟草感染24 h后出现DNA Laddering而转化烟草则没有;Western blotting结果显示PR-1蛋白表达没有显著差异。但接种另外一种病原真菌核盘菌(Sclerotinia sclerotiorum)后的RT-PCR分析结果表明,表达P35蛋白的烟草可增强感染核盘菌后PR-1基因的转录,而且表达时间提前。以上结果说明p35基因介导的广谱抗病反应的机理与接种的不同病原有关,对不同病原物的抗病机理存在差异,除抑制细胞凋亡外,还可能通过激活PR基因的表达提高对病原物的抗病能力。
Anti-apoptotic genes derived from insect viruses and animals can induce plant resistance to either biotic or abiotic stress, but their mechanisms of resistance are reported differently or even reversed. In the present study, the p35 gene derived from Autographa californica nucleopolyhedrovirus was transformed into tobacco. The expression of P35 protein was detected by Western blotting in T1 generation tobacco and the resistance effect of tobacco transformed with Tobacco mosaic virus (TMV) . Further studies on the mechanism of resistance showed that there was no significant difference between the transgenic and wild-type tobacco infected with TMV after induction of hydrogen peroxide accumulation. However, DNA Laddering occurred 24 h after wild-type tobacco infection and no tobacco was transformed. Western blotting showed that PR-1 protein expression No significant difference. However, RT-PCR analysis after inoculation of Sclerotinia sclerotiorum showed that the P35 protein-expressing tobacco enhanced the transcription of PR-1 gene after S. pneumoniae infection, and the expression time was earlier. The above results indicate that the mechanism of p35 gene-mediated broad-spectrum disease-resistance is related to the different pathogens inoculated and the pathogenic mechanisms of different pathogens are different. In addition to inhibiting apoptosis, it may also increase the expression of PR gene by activating PR Disease resistance of pathogens.