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Identification of pathogenicity genes regulated by two kinds of transcription regulators of the fungal plant pathogen Fusarium oxysporum

植物病原菌尖孢镰刀菌两种转录调控子致病基因的鉴定

基本信息

批准号：
15208005
负责人：
TSUGE Takashi
金额：
$ 33.03万
依托单位：
Nagoya University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (A)
财政年份：
2003
资助国家：
日本
起止时间：
2003 至 2006
项目状态：
已结题

项目摘要

We previously identified two genes, FOW2 and FOW3, which encode Zn(II)2Cys6-type and His2Cys2-type transcription regulators, respectively, essential for plant infection in the melon wilt pathogen Fusarium oxysporum f. sp. melonis. In this research, we attempted to isolate Fow2-or Fow3-regulated genes.We constructed cDNA subtraction libraries, which contained genes with changes in transcription levels caused by mutations in FOW2 and FOW3. Screening of these libraries by cDNA dot-blot differential hybridization and RNA gel blot analyses identified six genes positively regulated by Fow2. We made mutants of these genes and identified a Ren1-regulated pathogenicity gene, named VRF2-1. The vrf2-1 mutant showed reduced virulence on melon plants, although the fow2 mutant completely lost pathogenicity. This result indicates that Fow2 regulates the transcription of multiple pathogenicity genes, including VRF2-1. We sequenced a BAC clone encoding FOW2 and found that VRF2-1 is also encoded by this … More clone. This clone contains additional 29 genes, of which 10 were found to be possibly regulated by Ren1, suggesting that the Fow2-regulated genes are closely linked with FOW2. Database homology search and PCR analysis revealed that the filamentous ascomycetes species have the FOW2 homologues. This suggests that the FOW2 homologues also control plant infection in other fungi.We also identified two genes, REN1 and FoSTUA, which encode different types of transcription regulators essential for conidiation in F. oxysporum. In plant pathogenic fungi, conidia play important roles in the disease cycle as disseminated propagules and primary and secondary inocula. We performed expressed sequence tag analysis during vegetative growth and conidiation and identified 496 genes specifically detected in the conidiation cDNA library. Comparison of the transcription levels of the 496 genes between the wild-type strain and the ren1 mutant or the fostuA mutant identified 35 and three genes regulated by Ren1 and FoStuA, respectively. Less

我们以前确定了两个基因，FOW 2和FOW 3，这两个基因分别编码Zn（II）2Cys 6型和His 2Cys 2型转录调节因子，在甜瓜枯萎病病原体尖孢镰刀菌f.甜瓜（melonis）在本研究中，我们试图分离Fow 2或Fow 3调控基因，构建cDNA消减文库，其中包含FOW 2和FOW 3突变引起的转录水平变化的基因。通过cDNA斑点杂交差异杂交和RNA凝胶印迹分析筛选这些文库，鉴定出Fow 2正调控的6个基因。我们对这些基因进行了突变，并确定了Ren 1调节的致病基因，命名为VRF 2 -1。vrf 2 -1突变体对甜瓜的致病性降低，而fow 2突变体完全丧失致病性。这一结果表明，Fow 2调控包括VRF 2 -1在内的多个致病基因的转录。我们对编码FOW 2的BAC克隆进行了测序，发现VRF 2 -1也是由该基因编码的。 ...更多信息分身该克隆含有另外29个基因，其中10个被发现可能受Ren 1调控，这表明Fow 2调控的基因与FOW 2紧密连锁。数据库同源性搜索和PCR分析表明，丝状子囊菌物种具有FOW 2同源物。我们还鉴定了两个基因，REN 1和FoSTUA，它们编码不同类型的转录调节因子，对F.尖孢菌属在植物病原真菌中，分生孢子作为散播性繁殖体、初次接种体和二次接种体在病害循环中起着重要作用。我们在营养生长和分生孢子过程中进行了表达序列标签分析，并确定了496个基因在分生孢子cDNA文库中特异性检测。野生型菌株和ren 1突变体或fostuA突变体之间的496个基因的转录水平的比较确定了35和3个基因的Ren 1和FostuA，分别调节。少