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由小麦条锈菌(Puccinia striiformis f.sp.tritici,Pst)引起的小麦条锈病是我国小麦(Triticum aestivum)最严重的流行病害之一。中国小麦条锈菌表现出高度的遗传多样性和毒性变异,现已研究表明小麦条锈菌经有性生殖可发生致病性变异并产生新的小种。冬孢子作为小麦条锈菌有性阶段的开始,是小麦条锈菌生活史中完成有性生殖过程必不可少的阶段。为分析小麦条锈菌产冬孢子时期寄主植物的叶片总蛋白表达变化,探究寄主植物如何响应冬孢子产生,本研究利用三氯乙酸(trichloroacetic acid,TCA)/酚抽提法提取产冬孢子时期和同时期未接种的小麦叶片总蛋白,经双向电泳技术分离后,利用PDQuest软件进行分析并选取显著上调表达的蛋白进行Orbitrap质谱鉴定。结果表明,在产冬孢子时期的寄主叶片总蛋白图谱中发现22个蛋白点(上调表达大于1.5倍且符合t检验),经功能注释发现这些差异蛋白参与糖代谢、抗逆境以及植物衰老相关代谢途径。之后利用q RT-PCR技术对蛋白水平的结果进行验证,结果发现这22个蛋白点中有16个差异蛋白的基因在转录水平上调表达(相对表达倍数大于1.5倍),证实了蛋白结果的可靠性。本研究通过双向电泳技术发现小麦条锈菌冬孢子的产生影响了寄主叶片参与糖代谢、抗逆境及植物衰老相关代谢过程蛋白的表达,并推测小麦条锈菌冬孢子的产生可能与寄主植物的衰老有关。本研究为进一步深入了解小麦条锈菌冬孢子阶段寄主植物与病原菌的互作机制提供了一定的理论基础,对于揭示冬孢子的形成机制和全面了解小麦条锈菌的有性阶段有重要意义。
Wheat stripe rust caused by Puccinia striiformis f. Sp. Tritici (Pst) is one of the most serious epidemic diseases of Triticum aestivum in China. Chinese wheat stripe rust shows a high degree of genetic diversity and toxicity variation. It has been shown that the pathogenic mutation of wheat stripe rust occurs through sexual reproduction and produces new races. As a start of the sexual stage of wheat stripe rust, teliospore is an indispensable stage in the sexual reproduction of wheat stripe rust. In order to analyze the change of the total protein expression of the host plant during the winter spore stage of wheat stripe rust and explore how the host plant responds to the production of the teliospore. In this study, trichloroacetic acid (TCA) / phenol extraction The total protein of unvaccinated wheat leaves in the same period was separated by two-dimensional gel electrophoresis and analyzed by PDQuest software. The proteins that were significantly up-regulated were selected for Orbitrap mass spectrometry identification. The results showed that there were 22 protein spots (up-regulated more than 1.5 times and t-test) in the host protein map of the winter-producing spores. Functional differences indicated that these differential proteins were involved in glucose metabolism, stress tolerance and plant-related metabolism way. Afterwards, the result of q RT-PCR was used to verify the protein level. The results showed that 16 of the 22 protein spots were upregulated at the transcription level (relative expression multiple more than 1.5 times), which confirmed the reliable result Sex. In this study, it was found by two-dimensional electrophoresis that teliospore production in wheat stripe rust affected the expression of the host leaves involved in the metabolism of sugar, anti-stress and the metabolic processes involved in plant senescence, and speculated that the production of winter spores of wheat stripe rust may be related to the host plant Related to aging. This study provided a theoretical basis for further understanding of the mechanism of interaction between host plants and pathogenic fungi in the winter spore stage of wheat stripe rust and is of great significance for revealing the formation mechanism of winter spores and understanding the sexual stage of wheat stripe rust.