請用此 Handle URI 來引用此文件: http://hdl.handle.net/11455/95772
標題: 辣椒炭疽病之候選效應蛋白功能性分析
Functional Analysis of Candidate Effector Proteins of Colletotrichum acutatum strain 524
作者: 江政霖
Chen-Lin Chiang
關鍵字: 辣椒炭疽病
Colletotrichum acutatum
LysM domain
Colletotrichum acutatum
effector protein
hypothetical protein
LysM effector
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摘要: 辣椒炭疽病菌Colletotrichum acutatum在台灣危害辣椒及多種重要經濟作物。目前已知此病原菌以半活體營養生長策略來獲取寄主養分,但詳細的致病機制則很少有相關的探討。近年來我們已建立辣椒炭疽病菌強毒力菌株C. acutatum Coll-524侵染辣椒果實過程之轉錄體。於轉錄體分析的結果發現Coll-524在感染辣椒的過程中,會外泌一群小分子的蛋白質,這些小分子蛋白可能參與Coll-524於辣椒果實之半活體營養生長。我們從中挑選出13個Candidate Effector Proteins (CEPs),這13個CEPs蛋白質長度小於300個胺基酸、帶有訊息傳遞胜肽、功能尚未被研究及感染過程中於特定時期表現量最高。本研究進行其中2個CEPs功能的探討,為功能未知的CEP002以及帶有LysM domain的CEP012;另外也同時研究CEP012的旁系同源基因Ca_5620的功能。本研究使用基因剔除、過量表現及於菸草葉片中短暫大量表現等策略進行基因功能分析。將CEP002基因剔除後,進行致病力、生長與孢子發芽率等試驗分析,結果顯示CEP002 並未影響Coll-524 之上述功能,但若將CEP002持續大量表現於Coll-524,則可提升Coll-524對辣椒的致病力;在圓葉菸草葉片中表現CEP002進行誘導細胞死亡及抑制BAX所誘導的細胞死亡分析,並未觀察到顯著的差異。在CEP012與Ca_5620的功能研究,剔除CEP012或Ca_5620後,發現皆可造成Coll-524對辣椒之致病力下降。未來可針對持續表現CEP002是如何提升Coll-524致病力以及CEP012與Ca_5620同時存在於Coll-524基因體中的意義等方面進行探討。
Colletotrichum acutatum dramatically diminishes yield rates of chili peppers and other crops in Taiwan. This pathogen uses hemibiotrophic strategy for nutrient acquisition but how pathogen effectors alter plant immunity or responses remains unknown. In our lab, transcriptomic analysis of Coll-524 during pathogenesis on chili pepper fruit showed that a group of genes encoding small and secreted proteins expressed specifically during infection processes, called candidate effector proteins (CEPs). In this study, we analyzed the function of unknown effector CEP002 and two LysM domain-containing effector, CEP012 and its paralogous gene Ca_5620, by gene knockout and overexpression in Coll-524, and transient expression in Nicotiana benthamiana. Here, we found that knocking out CEP002 in Coll-524 genome did not affect on pathogenicity, mycelia growth and spore germination, but constitutive expression of CEP002-gfp in Coll-524 could increase fungal virulence to chili peppers. However, transient expression of CEP002 in N. benthamiana didn’t show any notable phenotype. For the functional analysis of CEP012 and Ca_5620, we found that knocking out eitheir CEP012 or Ca_5620 in Coll-524 genome would reduce the virulence of Coll-524 when inoculated on chili peppers. In the future, the reason that the constitutive expression of CEP002-gfp increases virulence of Coll-524 on chili peppers and the biological meaning of the two LysM effectors existing in Coll-524 genome remain to be investigated.
URI: http://hdl.handle.net/11455/95772
文章公開時間: 10000-01-01

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