本文關(guān)鍵詞： 玉米大斑病菌 轉(zhuǎn)錄因子 cAMP信號途徑 qRT-PCR　出處：《農(nóng)業(yè)生物技術(shù)學報》2017年10期 　論文類型：期刊論文

【摘要】：Flo8是cAMP信號途徑下游的重要轉(zhuǎn)錄因子,與真菌的生長發(fā)育密切相關(guān)。本研究旨在獲得玉米大斑病菌(Setosphaeria turcica)轉(zhuǎn)錄因子Flo8編碼基因,并分析其在病菌侵染結(jié)構(gòu)發(fā)育過程及病菌侵染寄主早期階段的表達模式,為深入探索該轉(zhuǎn)錄因子在病菌發(fā)育及致病過程中的功能奠定基礎(chǔ)。本研究以白色念珠菌(Candida albicans)Flo8氨基酸序列為探針在玉米大斑病菌數(shù)據(jù)庫中進行同源搜索,利用PCR技術(shù)對玉米大斑病菌轉(zhuǎn)錄因子Flo8的編碼基因進行克隆,通過GSDS、Prot Param、SOMPA和SMART在線軟件進行基因及其編碼產(chǎn)物結(jié)構(gòu)特征預(yù)測分析,利用WoLF PSORT分析蛋白的亞細胞定位;利用qRTPCR技術(shù)研究其在病菌侵染結(jié)構(gòu)形成過程中不同階段的轉(zhuǎn)錄水平。結(jié)果表明,玉米大斑病菌中存在1個含有LisH保守結(jié)構(gòu)域的蛋白,屬于典型的FLO8同源蛋白,將其編碼基因命名為StFlo8。該基因DNA全長為2 384 bp,c DNA全長為2 037 bp,含有6個內(nèi)含子,7個外顯子,編碼678個氨基酸。表達規(guī)律分析結(jié)果表明,與菌絲時期相比,StFlo8在孢子時期及芽管時期表達量顯著降低,在附著胞時期和侵染絲時期表達量顯著升高,達到菌絲時期的4倍以上;而在侵染感病寄主葉片過程中,StFlo8表達量呈現(xiàn)先下調(diào)后上調(diào)再下調(diào)的趨勢,并在接種后18 h表達量達到最高,約為接種初期的2倍。以上結(jié)果表明StFlo8參與調(diào)控病菌附著胞發(fā)育及侵染絲形成,該結(jié)果為進一步研究StFlo8基因的功能及玉米大斑病菌的侵染機制提供了理論依據(jù)。
[Abstract]:Flo8 is an important transcription factor downstream of cAMP signaling pathway. The aim of this study was to obtain the transcription factor Flo8 coding gene of Setosphaeria turcica. The expression patterns of the pathogen in the early stage of infection and host were analyzed. In order to further explore the function of the transcription factor in the development and pathogenicity of Candida albicans. in this study, Candida albicans. The amino acid sequence of Flo8 was homologous searched in the database of Corn spot fungus. The coding gene of Flo8 was cloned by PCR technique. SOMPA and SMART software were used to predict the structural characteristics of genes and their encoded products, and WoLF PSORT was used to analyze the subcellular localization of proteins. QRTPCR technique was used to study the transcription level in different stages of infection structure formation. The results showed that there was a protein containing the conserved domain of LisH in the pathogen. It is a typical FLO8 homologous protein, and its coding gene is named StFlo8. The DNA length of the gene is 2 384 BP, and the length of DNA is 2 037 BP. It contains 6 introns and 7 exons and encodes 678 amino acids. The results of expression analysis showed that the expression of StFlo8 in spore stage and bud tube stage was significantly lower than that in hyphae stage. The expression level increased significantly at the appressant stage and the infection stage, and reached more than 4 times of the mycelium stage. The expression of StFlo8 in infected host leaves showed a tendency of down-regulation, then up-regulation and down-regulation, and reached the highest level at 18 h after inoculation. The results showed that StFlo8 was involved in the development of appressorium and the formation of infected filaments. The results provided a theoretical basis for the further study of the function of StFlo8 gene and the infection mechanism of Corn spot fungus.
【作者單位】： 河北農(nóng)業(yè)大學生命科學學院/河北省植物生理與分子病理學重點實驗室;
【基金】：國家自然科學基金(No.31301616和No.31601598) 河北省自然科學基金(No.C2016204160) 河北省高等學�？茖W技術(shù)研究項目(No.QN2016071)
【分類號】：S435.131.4
【正文快照】： *同等貢獻作者玉米大斑病菌(Setosphaeria turcica)是一種侵染玉米(Zea mays)的重要絲狀病原真菌,其引起的玉米大斑病(northern corn leaf blight)在世界低溫潮濕地域分布廣泛,對玉米生產(chǎn)造成巨大的經(jīng)濟損失(宋文靜等,2008)。由于該病菌生理分化明顯,容易變異,近年來由其引發(fā)

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