A MAP kinase gene, Clk1, is required for conidiation and pathogenicity in the phytopathogenic fungus Curvularia lunata
A MAP kinase gene, Clk1, is required for conidiation and pathogenicity in the phytopathogenic fungus Curvularia lunata
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DOI：
10.1002/jobm.201100518
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发表时间：
2013-03
影响因子：
3.1
通讯作者：
Shigang Gao;Feihong Zhou;Tong Liu;Y. Li;Jie Chen
中科院分区：
文献类型：
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作者：
Shigang Gao;Feihong Zhou;Tong Liu;Y. Li;Jie Chen
丝裂原活化蛋白激酶(MAPK)级联通路是高度保守的信号转导通路，在细胞外信号转导中发挥着广泛的重要作用。根据已知MAPK基因的保守区设计了一对引物，通过聚合酶链式反应的方法分离到了玉米弯孢霉CLK1的第一个MAPK基因。Southern杂交分析表明，该基因以单拷贝形式存在于基因组中。预测的氨基酸序列(352个氨基酸)与其他植物病原真菌的MAP激酶高度同源。通过RACE和基因组行走技术获得CLK1基因的侧翼区域。为了了解CLK1在新月弯曲霉中的作用，采用定向基因阻断的方法构建了CLK1突变体。研究发现，缺乏CLK1功能结构域的突变体不能产生分生孢子，但倾向于形成一些特殊的厚壁孢子形状的结构。CLK1突变体在恶劣环境(24℃)中生长缓慢，产生的细胞降解酶(CWDEs)比野生型少，几乎不能通过人工伤口侵染玉米叶片。
Mitogen‐activated protein kinase (MAPK) cascades are highly conserved signal transduction pathways, which play a wide variety of important roles in extracellular signal transduction. The first MAPK gene of the maize pathogen Curvularia lunata, Clk1, was isolated via a PCR‐based approach with a primer pair designed on the basis of conserved regions of known MAPKs. Southern blot analysis showed that the gene existed in the genome as a single copy. The predicted amino acid sequence (352 amino acids) was highly homologous with MAP kinases of other phytopathogenic fungi. Flanking regions of Clk1 were obtained through RACE and genomic walking technology. To understand the role of Clk1 in C. lunata, targeted gene disruption was adopted to construct Clk1 mutants. It was found that mutants lacking functional domain of Clk1 were not able to produce conidia but tended to form a few special chlamydospore‐shaped structures. Clk1 mutants grew slower in adverse environments (at 24°C), produced less cell degrading enzymes (CWDEs) than the wild type, and they were almost unable to infect maize leaves via artificial wounds.