本文選題：灰葡萄孢 + 致病力　； 參考：《華中農(nóng)業(yè)大學(xué)》2017年碩士論文

【摘要】：灰葡萄孢(Botrytis cinerea)是一種世界范圍內(nèi)廣泛分布的死體營養(yǎng)型病原真菌,是灰霉病的主要病原菌,寄主范圍十分廣泛�；移咸焰吲c核盤菌親緣關(guān)系較近,但盾殼霉不能寄生灰葡萄孢而能寄生與蠶豆葡萄孢屬親緣性較近的HN454菌株,探究灰葡萄孢中抵制盾殼霉寄生的基因功能至關(guān)重要。通過盾殼霉分別與灰葡萄孢、核盤菌、HN454菌株侵染互作的RNA-seq數(shù)據(jù),分析篩選出了顯著差異表達(dá)的BC1G_00904、BC1G_12653、BC1G_03604和BC1G_00245。本研究利用基因敲除和互補(bǔ)技術(shù)獲得相應(yīng)的突變體,并探究基因功能。BC1G_00904含有高度保守的Mito-carr(Mitochondrial Carrier)結(jié)構(gòu)域,預(yù)測編碼蛋白功能與羧酸鹽運(yùn)輸相關(guān),命名為BcMito-TTP。釀酒酵母(Saccharomyces cerevisiae)中研究發(fā)現(xiàn)該羧酸鹽轉(zhuǎn)運(yùn)蛋白主要負(fù)責(zé)催化檸檬酸鹽通過線粒體內(nèi)膜流出以交換羧酸鹽H+離子等。較出發(fā)菌株相比,BC1G_00904敲除突變體在致病力與產(chǎn)孢量方面顯著降低,并延緩了菌核的形成,H2O2含量顯著降低,推測ROS是致病力降低的主要原因,同時也驗(yàn)證出基因功能與羧酸鹽的運(yùn)輸有明顯關(guān)系。BC1G_12653和BC1G_00245保守結(jié)構(gòu)域分別屬于MFS(Major Faciltator Superfamily)和GHS(Glyco_hydro_92 superamily),BC1G_03604為假定蛋白。實(shí)驗(yàn)發(fā)現(xiàn)BC1G_12653敲除突變體菌落形態(tài)無明顯差異,但生長速度減慢、產(chǎn)孢量與致病力明顯降低,同時能被盾殼霉輕微寄生。BC1G_03604敲除致病力顯著減弱,菌核數(shù)量有所減少。BC1G_00245基因?qū)移咸焰呱飳W(xué)性狀無影響。研究結(jié)果有助于我們理解灰葡萄孢的致病、孢子和菌核形成的分子機(jī)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.
【學(xué)位授予單位】：華中農(nóng)業(yè)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：S432.44

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