本文選題：灰葡萄孢菌　切入點(diǎn)：隔膜蛋白septin　出處：《吉林大學(xué)》2017年碩士論文

【摘要】：灰葡萄孢菌是一種全球重要的植物病原真菌,屬于典型的死體營(yíng)養(yǎng)型,寄主范圍十分廣泛,能夠侵染多達(dá)1400種植物。該菌不僅能在植物生長(zhǎng)期侵染,也能在果蔬儲(chǔ)藏期侵染發(fā)病,危害廣泛,損失嚴(yán)重。目前灰葡萄孢菌作為一種模式植物病原備受關(guān)注,但關(guān)于其寄主侵染過(guò)程的分子機(jī)理仍不清楚。在過(guò)往的研究中發(fā)現(xiàn),灰葡萄孢菌需要形成特定的侵染結(jié)構(gòu):附著胞或侵染墊,才能正常侵入寄主,進(jìn)一步分泌多種致病因子殺死寄主從而獲取營(yíng)養(yǎng)。此外,隔膜蛋白septin作為真核生物中一類(lèi)高度保守的蛋白家族,參與細(xì)胞骨架,細(xì)胞周期,細(xì)胞分泌等多項(xiàng)生命過(guò)程并發(fā)揮重要功能。在真菌中septin蛋白家族具有四個(gè)核心成員:Sep3、Sep4、Sep5、Sep6,其中Sep4是最小的蛋白。本研究重點(diǎn)解析新型細(xì)胞骨架隔膜蛋白septin如何在灰葡萄孢菌生長(zhǎng)發(fā)育發(fā)揮功能,并參與寄主侵染過(guò)程,主要研究結(jié)果如下:(1)通過(guò)生物信息學(xué)分析,檢索到灰葡萄孢菌中四個(gè)核心septins組分:BcSep3、BcSep4、BcSep5、BcSep6;(2)利用農(nóng)桿菌介導(dǎo)轉(zhuǎn)化,同源重組基因替換的方法,篩選獲得灰葡萄孢菌四個(gè)核心septin基因敲除突變體ΔBcsep3,ΔBcsep4,ΔBcsep5,ΔBcsep6菌株。通過(guò)致病性分析,結(jié)果表明核心septins組分是灰葡萄孢菌致病所必需的;(3)通過(guò)野生型與突變體表型差異分析,發(fā)現(xiàn)BcSep4是灰葡萄孢菌侵染結(jié)構(gòu)發(fā)育所必需的;另外,BcSep4還在灰葡萄孢菌無(wú)性繁殖、芽管發(fā)育、柔性生長(zhǎng)、逆境響應(yīng)、胞內(nèi)物質(zhì)運(yùn)輸?shù)冗^(guò)程中發(fā)揮重要功能;(4)通過(guò)GFP融合亞細(xì)胞定位,證明BcSep4可能通過(guò)參與脂膜微曲面形成,調(diào)控灰葡萄孢菌生長(zhǎng)發(fā)育和侵染結(jié)構(gòu)形成過(guò)程;(5)利用二亞苯基碘(diphenylene iodonium,DPI)和環(huán)磷酸腺苷(cyclic adenosine monophosphate,cAMP)處理,發(fā)現(xiàn)灰葡萄孢菌BcSep4在侵染結(jié)構(gòu)形成過(guò)程中受活性氧ROS信號(hào)傳導(dǎo)調(diào)控,并與cAMP信號(hào)傳導(dǎo)有關(guān);(6)通過(guò)對(duì)四個(gè)核心septin基因缺失突變體菌株ΔBcsep3、ΔBcsep4、ΔBcsep5、ΔBcsep6生物表型分析,發(fā)現(xiàn)四個(gè)核心septin組分都是灰葡萄孢菌侵染結(jié)構(gòu)形成所必需的;(7)對(duì)四個(gè)核心septin組分分別進(jìn)行亞細(xì)胞定位,結(jié)果表明BcSep3、BcSep4、BcSep5、BcSep6在胞內(nèi)能夠以不同的自我組裝分配模式,影響細(xì)胞極性,共同調(diào)控灰葡萄孢菌生長(zhǎng)發(fā)育和侵染結(jié)構(gòu)形成。本研究首次解析灰葡萄孢菌septin的生物學(xué)功能,讓我們對(duì)灰葡萄孢菌的生長(zhǎng)發(fā)育和分子致病機(jī)理有了更深入的認(rèn)識(shí)。由于植物中沒(méi)有septin存在,因此未來(lái)可能作為潛在的防治靶標(biāo)應(yīng)用于農(nóng)業(yè)防治中。
[Abstract]:Grapevine cinerea is one of the most important plant pathogenic fungi in the world. It belongs to the typical dead body nutrition type and can infect up to 1400 species of plants in a wide range of hosts. It can also infect fruits and vegetables during storage period, causing extensive damage and serious losses. At present, Grapevine Aspergillus, as a model plant pathogen, has attracted much attention, but the molecular mechanism of its host infection process is still unclear. Grapevine fungus needs to form a specific infection structure: appressorium or infection pad in order to normally invade the host, further secreting a variety of pathogenic factors to kill the host and obtain nutrition. As a family of highly conserved proteins in eukaryotes, membrane protein septin is involved in cytoskeleton and cell cycle. In fungi, the septin protein family has four core members: Sep3C Sep4N Sep5N Sep6, in which Sep4 is the smallest protein. This study focuses on how the novel cytoskeleton membrane protein septin is present in the grapevine grays of Staphylococcus gravis. The main purpose of this study is to investigate the role of septin in the development of a novel cytoskeleton membrane protein septin. The growth and development of grape spores play a role, The main results were as follows: (1) by bioinformatics analysis, we found four core septins components of Grapevine Aspergicus: BcSep3, BcSep4CSep4BcSep5BcSep5BcSep6 / 2) by Agrobacterium tumefaciens mediated transformation and homologous recombination gene replacement. Four core septin gene knockout mutants 螖 Bcsep3, 螖 Bcsep4, 螖 Bcsep5, 螖 Bcsep6 were obtained. The results of pathogenicity analysis showed that the core septins components were essential for virulence of grapevine. It was found that BcSep4 was necessary for the development of the infective structure of Grapevine Aspergillus, and that BcSep4 was also used in asexual reproduction, bud tube development, flexible growth and stress response of Grapevine. Through GFP fusion subcellular localization, it is proved that BcSep4 may be involved in the formation of lipid membrane microsurfaces. In order to regulate the growth and development of grapevine and the formation of infection structure, we found that BcSep4 was regulated by active oxygen species (ROS) signal transduction during the process of infection structure formation by using diphenyl iodiphenylene iodoniumn (DPI) and cyclic adenosine monophosphate (cAMP), which were treated with diphenyl iodiphenylene and cyclic adenosine monophosphate monophosphate (cAMP). Four core septin gene deletion mutants 螖 Bcsep3, 螖 Bcsep4, 螖 Bcsep5, 螖 Bcsep6 were analyzed. It was found that the four core septin components were subcellular localization of the four core septin components, which were necessary for the formation of infection structure of grapevine. The results showed that BcSep3, BcSep4, BcSep4 and BcSep5BcSep6 could affect cell polarity in different self-assembly patterns. In this study, the biological function of septin was analyzed for the first time. As there is no septin in plants, it may be used as a potential target for agricultural control in the future.
【學(xué)位授予單位】：吉林大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類(lèi)號(hào)】：S432.44

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相關(guān)期刊論文 前2條

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本文編號(hào)：1659269