Kwon Sun-Jung, Cho Sang-Yun, Lee Kyung-Mi, Yu Jisuk, Son Moonil, Kim Kook-Hyung
Research Institute for Agriculture and Life Sciences, Seoul National University, Seoul 151-921, Republic of Korea.
Virus Res. 2009 Sep;144(1-2):96-106. doi: 10.1016/j.virusres.2009.04.004. Epub 2009 Apr 15.
Fusarium graminearum virus-DK21 (FgV-DK21), which infects the plant pathogenic F. graminearum, perturbs host developmental processes such as sporulation, morphology, pigmentation, and attenuates the virulence (hypovirulence) of the host. To identify the differentially expressed F. graminearum proteins by FgV-DK21 infection, we have used two-dimensional electrophoresis with mass spectrometry using proteins extracted from virus-free and FgV-DK21-infected strains. A total of 148 spots showing an altered expression were identified by PDQuest program. Among these spots, 33 spots were exclusively analyzed including 14 spots from FgV-DK21-infected and 19 spots from virus-free strains by ESI-MS/MS analyses and successfully identified 23 proteins. Seven proteins including sporulation-specific gene SPS2, triose phosphate isomerase, nucleoside diphosphate kinase, and woronin body major protein precursor were induced or significantly up-regulated by FgV-DK21 infection. A significant decrease or down regulation of 16 proteins including enolase, saccharopine dehydrogenase, flavohemoglobin, mannitol dehydrogenase and malate dehydrogenase caused by FgV-DK21 infection was also identified. Variations of protein expression were also further investigated at the mRNA level by real-time RT-PCR analysis, which confirmed the proteomic data for 9 out of the representative 11 selected proteins including 5 proteins from up-regulated group and 6 proteins from down-regulated group. Further investigation of these differentially expressed proteins will provide novel insights into the molecular responses of F. graminearum to FgV-DK21 infection.
禾谷镰刀菌病毒-DK21(FgV-DK21)感染植物病原性禾谷镰刀菌,扰乱宿主的发育过程,如孢子形成、形态、色素沉着,并减弱宿主的毒力(低毒力)。为了鉴定受FgV-DK21感染后禾谷镰刀菌中差异表达的蛋白质,我们使用二维电泳结合质谱分析,从无病毒和FgV-DK21感染的菌株中提取蛋白质。通过PDQuest程序共鉴定出148个表达发生改变的斑点。在这些斑点中,通过电喷雾串联质谱分析专门分析了33个斑点,包括14个来自FgV-DK21感染菌株的斑点和19个来自无病毒菌株的斑点,并成功鉴定出23种蛋白质。包括孢子形成特异性基因SPS2、磷酸丙糖异构酶、核苷二磷酸激酶和沃罗宁体主要蛋白前体在内的7种蛋白质在FgV-DK21感染后被诱导或显著上调。还鉴定出由FgV-DK21感染导致的16种蛋白质的显著减少或下调,包括烯醇化酶、酵母氨酸脱氢酶、黄素血红蛋白、甘露醇脱氢酶和苹果酸脱氢酶。还通过实时逆转录聚合酶链反应分析在mRNA水平进一步研究了蛋白质表达的变化,这证实了11种代表性选择蛋白质中的9种蛋白质组学数据,包括上调组的5种蛋白质和下调组的6种蛋白质。对这些差异表达蛋白质的进一步研究将为禾谷镰刀菌对FgV-DK21感染的分子反应提供新的见解。