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为明确毁灭炭疽菌Colletotrichum destructivum诱抗蛋白诱导烟草的抗病性及其作用,采用喷雾、摩擦接种方法及RT-PCR技术研究了诱抗蛋白的预防保护作用,以及烟草悬浮细胞经诱导后过氧化物酶(POD)、多酚氧化酶(PPO)、苯丙氨酸解氨酶(PAL)活性及脯氨酸(Pro)含量和病程相关基因表达的变化。结果表明,接种3、5和7 d后,该诱抗蛋白对烟草炭疽病的诱抗效果分别为58.00%、48.99%和49.65%,对烟草白粉病的诱抗效果分别为83.26%、80.76%和78.60%,并可以抑制烟草普通花叶病毒的复制及在寄主体内的扩增;经诱抗蛋白处理后,烟草悬浮细胞POD、PPO、PAL活性及Pro含量明显提高;诱抗蛋白能够诱导烟草病程相关蛋白基因PR-1a、PR-1b以及抗病信号传导途径关键基因NPR1的表达。表明毁灭炭疽菌诱抗蛋白可诱导烟草产生抗病性,可能与烟草悬浮细胞中POD、PAL、PPO的活性及Pro的含量提高以及相关病程基因表达有关。
In order to clarify the anti-pathogenicity and the role of Colletotrichum destructivum induced by anthrax-induced anti-protein in tobacco, the preventive and protective effects of anti-induced protein were studied by spraying, frictional inoculation and RT-PCR, (POD), polyphenol oxidase (PPO), phenylalanine ammonia lyase (PAL) activity and proline (Pro) content and disease-related gene expression changes. The results showed that the induced resistance to tobacco anthracnose was 58.00%, 48.99% and 49.65% respectively at 3, 5 and 7 days after inoculation, and the induced resistance to tobacco powdery mildew was 83.26% and 80.76% And 78.60%, respectively, and could inhibit the replication of tobacco mosaic virus and its amplification in the host tobacco. The POD, PPO, PAL activity and Pro content of the tobacco suspension cells were significantly increased after the induced anti-protein treatment. PR-1a, PR-1b, and NPR1, a key gene in disease-resistance signal transduction pathways. The results showed that the anti-anthrax-induced protein could induce tobacco-induced disease resistance, which may be related to the activity of POD, PAL and PPO in tobacco suspension cells and the increase of Pro content and the gene expression of related pathogens.