盾壳霉诱导的灰葡萄孢基因功能研究

发布时间：2018-05-14 17:14

本文选题：灰葡萄孢 + 致病力　；参考：《华中农业大学》2017年硕士论文

【摘要】：灰葡萄孢(Botrytis cinerea)是一种世界范围内广泛分布的死体营养型病原真菌,是灰霉病的主要病原菌,寄主范围十分广泛。灰葡萄孢与核盘菌亲缘关系较近,但盾壳霉不能寄生灰葡萄孢而能寄生与蚕豆葡萄孢属亲缘性较近的HN454菌株,探究灰葡萄孢中抵制盾壳霉寄生的基因功能至关重要。通过盾壳霉分别与灰葡萄孢、核盘菌、HN454菌株侵染互作的RNA-seq数据,分析筛选出了显著差异表达的BC1G_00904、BC1G_12653、BC1G_03604和BC1G_00245。本研究利用基因敲除和互补技术获得相应的突变体,并探究基因功能。BC1G_00904含有高度保守的Mito-carr(Mitochondrial Carrier)结构域,预测编码蛋白功能与羧酸盐运输相关,命名为BcMito-TTP。酿酒酵母(Saccharomyces cerevisiae)中研究发现该羧酸盐转运蛋白主要负责催化柠檬酸盐通过线粒体内膜流出以交换羧酸盐H+离子等。较出发菌株相比,BC1G_00904敲除突变体在致病力与产孢量方面显著降低,并延缓了菌核的形成,H2O2含量显著降低,推测ROS是致病力降低的主要原因,同时也验证出基因功能与羧酸盐的运输有明显关系。BC1G_12653和BC1G_00245保守结构域分别属于MFS(Major Faciltator Superfamily)和GHS(Glyco_hydro_92 superamily),BC1G_03604为假定蛋白。实验发现BC1G_12653敲除突变体菌落形态无明显差异,但生长速度减慢、产孢量与致病力明显降低,同时能被盾壳霉轻微寄生。BC1G_03604敲除致病力显著减弱,菌核数量有所减少。BC1G_00245基因对灰葡萄孢生物学性状无影响。研究结果有助于我们理解灰葡萄孢的致病、孢子和菌核形成的分子机理。
[Abstract]:Botrytis cinerea is a widely distributed dead body nutritional pathogenic fungus in the world. It is the main pathogen of grey mold, and its host range is very wide. The relationship between Grapevine spores and Sclerotinia sclerotiorum is close, but the HN454 strains with close affinity to Vicia faba can not be parasitized by C. auricularia. It is very important to explore the gene function of resistance to the parasitism of C. lucifera in the spores of Grapevine. Based on the RNA-seq data of the interaction of C. capsicum and HN454, the BC1G00904 BC1G12653BC1G03604 and BC1G002405 were selected. In this study, the corresponding mutants were obtained by gene knockout and complementation techniques, and the gene function. BC1GSC00904 contained a highly conserved Mito-carr(Mitochondrial Carrier domain. The predicted protein function was related to carboxylate transport and was named BcMito-TTP. It was found in Saccharomyces cerevisiaeae that the carboxylate transporter was mainly responsible for catalyzing the outflow of citrate through mitochondria to exchange carboxylate H ions and so on. Compared with the original strain, the pathogenicity and sporulation capacity of the mutant BC1G\ +\% ROS\% ROS\% ROS\%? It was also confirmed that the gene function was significantly related to the transport of carboxylate. BC1G12653 and BC1G_00245 conserved domains belonged to MFS(Major Faciltator Superfamilyand GHS(Glyco_hydro_92 supermilyn BC1G03604 were assumed proteins. The results showed that there was no significant difference in colony morphology of BC1G_12653 knockout mutants, but the growth rate was slow, the sporulation and pathogenicity were significantly decreased, and the pathogenicity of BC1G_12653 knockout mutant. BC1G03604 was significantly weakened. The number of sclerotia decreased. BC1GST00245 gene had no effect on the biological characters of grapevine. The results help us to understand the molecular mechanism of pathogenic, spore and sclerotia formation.
【学位授予单位】：华中农业大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S432.44

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