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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

我们以前在甜瓜枯萎病病原菌上发现了两个基因FOW2和FOW3，它们分别编码锌(II)2Cys6型和His2Cys2型转录调控因子，它们对甜瓜枯萎病菌的侵染是必不可少的。甜瓜。在这项研究中，我们试图分离Fow2或Fow3调控的基因，我们构建了包含FOW2和FOW3突变导致转录水平变化的基因的cDNA消减文库。通过cDNA斑点杂交、差异杂交和RNA凝胶印迹分析对文库进行筛选，鉴定出6个受Fow2正调控的基因。我们对这些基因进行了突变，并鉴定出一个受Ren1调控的致病基因，命名为VRF2-1。Vrf2-1突变体对甜瓜植株的致病力降低，而fow2突变体则完全丧失了致病力。这一结果表明，Fow2调控包括VRF2-1在内的多个致病基因的转录。我们对编码FOW2的BAC克隆进行了测序，发现VRF2-1也由这个…编码更多的克隆人。该克隆包含另外29个基因，其中10个可能受Ren1调控，这表明Fow2调控的基因与FOW2密切相关。数据库同源性搜索和聚合酶链式反应分析表明，丝状子囊菌具有FOW2同源物。我们还鉴定了两个基因，REN1和FoSTUA，它们编码不同类型的转录调节因子，对尖孢镰刀菌的分生至关重要。在植物病原真菌中，分生孢子作为传播性繁殖和一次、二次接种，在病原菌的侵染过程中起着重要作用。我们进行了营养生长和分生过程中的表达序列标签分析，鉴定了在分生孢子文库中特异检测到的496个基因。将野生型菌株与ren1突变体和fostuA突变体之间的496个基因的转录水平进行比较，发现分别有35个基因和3个基因受到Ren1和FoStuA的调控。较少