Hou Yiping, Zheng Zhitian, Xu Shu, Chen Changjun, Zhou Mingguo
College of Plant Protection, Nanjing Agricultural University, Nanjing 210095, China; Key Laboratory of Monitoring and Management of Crop Diseases and Pest Insects, Ministry of Education, Nanjing 210095, China; Key Laboratory of Pesticide, Jiangsu Province, Nanjing 210095, China.
College of Plant Protection, Nanjing Agricultural University, Nanjing 210095, China; Key Laboratory of Monitoring and Management of Crop Diseases and Pest Insects, Ministry of Education, Nanjing 210095, China; Key Laboratory of Pesticide, Jiangsu Province, Nanjing 210095, China.
Pestic Biochem Physiol. 2013 Sep;107(1):86-92. doi: 10.1016/j.pestbp.2013.05.009. Epub 2013 May 20.
JS399-19 (2-cyano-3-amino-3-phenylancryic acetate), a novel cyanoacrylate fungicide, has powerful inhibition against Fusarium species, especially to Fusarium graminearum. Treated with JS399-19, mycelium of F. graminearum was distorted and swelled. The embranchment increased. In order to investigate the effect of JS399-19 on protein expression of F. graminearum, total protein of F. graminearum cultured in normal condition and that treated with 0.5 μg/mL (EC90 value) JS399-19 were extracted respectively and proteomic analysis was performed using two-dimensional gel electrophoresis. The expression levels of 38 proteins varied quantitatively at least twofold. 33 proteins out of the 38 were successfully identified by MALDI-TOF-MS/MS and MASCOT. According to the classification of physiological functions from Conserved Domain Database analysis, 19, 5, 2, 3, 2 and 2 proteins were respectively associated with metabolism, regulation, motility, defense, signal transduction, and unknown function, which indicated that energy metabolism, the synthesis and transport of proteins and DNA of F. graminearum were inhibited by JS399-19 in different degrees. The expression levels of the genes were further confirmed by quantitative real-time PCR analyses. This study represents the first proteomic analysis of F. graminearum treated by JS399-19 and will provide some useful information to find the mode of action of the fungicide against F. graminearum.
JS399-19（2-氰基-3-氨基-3-苯基丙烯酸乙酯）是一种新型氰基丙烯酸酯类杀菌剂，对镰刀菌属具有强大的抑制作用，尤其对禾谷镰刀菌。用JS399-19处理后，禾谷镰刀菌的菌丝体发生扭曲和肿胀，分支增多。为了研究JS399-19对禾谷镰刀菌蛋白质表达的影响，分别提取了在正常条件下培养的禾谷镰刀菌以及用0.5μg/mL（EC90值）JS399-19处理后的禾谷镰刀菌的总蛋白，并使用二维凝胶电泳进行蛋白质组学分析。38种蛋白质的表达水平在数量上至少有两倍的变化。这38种蛋白质中的33种通过基质辅助激光解吸电离飞行时间串联质谱（MALDI-TOF-MS/MS）和MASCOT软件成功鉴定。根据保守结构域数据库分析的生理功能分类，19、5、2、3、2和2种蛋白质分别与代谢、调节、运动、防御、信号转导和未知功能相关，这表明JS399-19在不同程度上抑制了禾谷镰刀菌的能量代谢、蛋白质和DNA的合成与转运。通过定量实时PCR分析进一步证实了这些基因的表达水平。本研究是对经JS399-19处理的禾谷镰刀菌的首次蛋白质组学分析，将为寻找该杀菌剂对禾谷镰刀菌的作用模式提供一些有用信息。
