Identification of Clt-1-Regulated Proteins Associated with the Production of Non-Host-Specific Toxin and Pathogenicity in Cochliobolus lunatus
Identification of Clt-1-Regulated Proteins Associated with the Production of Non-Host-Specific Toxin and Pathogenicity in Cochliobolus lunatus
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DOI：
10.1111/jph.12278
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发表时间：
2015-01-01
影响因子：
1.5
通讯作者：
Chen, Jie
中科院分区：
文献类型：
--
作者：
Gao, Jin-Xin;Jing, Jing;Chen, Jie
弯孢菌叶斑病是由弯孢菌（Curvularia lunata）（远形：Cochliobolus lunatus）引起的，它产生一种非宿主特异性毒素，称为5-（羟甲基）呋喃-2-羧酸甲酯（M5 HF 2C）。成功克隆了与M5 HF 2C产生和致病性密切相关的病原菌Clt-1基因。然而，与毒素产生或致病性相关的蛋白未被鉴定。本研究利用双向凝胶电泳（2-DE）和质谱（MS）技术，对新月形对虾野生型CX-3和Clt-1缺失突变体Clt-1中与Clt-1表达相关的特异性蛋白进行了鉴定。在Clt-1中鉴定了五种上调和两种下调的蛋白质。琥珀酸脱氢酶黄素蛋白亚基（Sdh 1 p）是琥珀酸脱氢酶的辅酶，参与毒素的解毒作用。真核细胞延伸因子3（EF-3）促进新月形念珠菌产生毒素所需的重要蛋白质的合成。黑色素合成相关蛋白scytalone酶（SCD）和应激耐受相关蛋白（HSP 30和HSP 70）有助于C. lunatus的致病性。实时荧光定量PCR（RT-qPCR）分析表明，Scd基因在转录水平上表现出相似的表达模式，与2-DE检测到的蛋白相对应。生物信息学分析表明，Scd基因编码一个188个氨基酸的scytalone水解酶亚基，在系统发育树中与dothideomycete SCD同源物聚为一个分支。关于Scd基因的确切作用的进一步研究正在进行中。
The Curvularia leaf spot is caused by Curvularia lunata (telemorph: Cochliobolus lunatus), which produces a non-host-specific toxin known as methyl 5-(hydroxymethyl) furan-2-carboxylate (M5HF2C). The Clt-1 gene, which is from a pathogen closely associated with M5HF2C production and pathogenicity, was successfully cloned. However, the proteins related to toxin production or pathogenicity were not identified. In this study, a proteomic approach based on two-dimensional gel electrophoresis (2-DE) and mass spectrometry (MS) techniques was used to identify the specific proteins associated with Clt-1 expression in C.lunatus wild-type CX-3 and Clt-1 deletion mutant Clt-1. Five upregulated and two downregulated proteins were identified in Clt-1. Succinate dehydrogenase flavoprotein subunit (Sdh1p) functions as a coenzyme of dehydrogenase in the anabolism of toxin. Eukaryotic elongation factor 3 (EF-3) facilitates the synthesis of important proteins required for the production of toxins in C.lunatus. Melanin synthesis-related protein scytalone dehydratase (SCD) and stress tolerance-related proteins (HSP30 and HSP70) contribute to the pathogenicity of C.lunatus. Real-time quantitative PCR (RT-qPCR) analysis showed that the Scd gene exhibited similar expression pattern at the transcriptional level, corresponding to the protein detected by 2-DE. Bioinformatics analysis showed that Scd encodes a scytalone dehydratase subunit of 188 amino acids, which is clustered into one branch with dothideomycete SCD homologs in a phylogenetic tree. Further study on the exact role of the Scd gene is underway.