iTRAQ蛋白质组学分析揭示西瓜响应CGMMV胁迫的蛋白质

iTRAQ-Based Proteomic Analysis Reveal Proteins Responsive to Cucumber Green Mottle Mosaic Virus Infection in Watermelon

黄瓜绿斑驳花叶病毒病(CGMMV)是一种危害西瓜生产的检疫性病害。为了在蛋白质水平研究西瓜对CGMMV的胁迫应答,本研究采用同位素标记相对和绝对定量(iTRAQ)技术对CGMMV接种前后的西瓜叶片进行蛋白质组学分析,获得参与CGMMV胁迫应答的差异表达蛋白质(DEPs),并对DEPs进行KEGG功能分析;利用Cytoscape构建蛋白质-蛋白质互作(PPI)网络;应用实时荧光定量PCR(RT-qPCR)验证蛋白质在CGMMV胁迫下的表达。结果共获得61个在CGMMV胁迫下的DEPs,包括核糖体蛋白、光系统PSⅠ和PSⅡ蛋白、ATP合成酶、NADH脱氢酶、热激蛋白HSP22.8、真核细胞翻译起始因子、蔗糖合成酶和乙烯响应因子等;KEGG功能分析表明DEPs参与光合作用、次生代谢物质生物合成和植物激素信号转导等代谢途径;PPI分析显示,48 h_0 h、25 d_0 h和25 d_48 h分别有13、32和43个DEPs在CGMMV胁迫下参与已知的蛋白互作网络;RT-qPCR获得了SUS4、RPL22、NAD7、PSBH、Defensin-like protein 5、ATPF、LSP1和HSP22.8在CGMMV不同侵染阶段的表达模式。本研究结果获得了西瓜响应CGMMV胁迫应答的蛋白质,为后续明确西瓜响应CGMMV的分子机理奠定了一定基础。

Cucumber green mottle mosaic virus(CGMMV) is a quarantine disease that affects watermelon production. To investigate the CGMMV-associated defense response in watermelon at proteomic level, we sampled watermelon leaves before and post CGMMV inoculation and employed isobaric tag for relative and absolute quantification(iTRAQ) technology to identify the differentially expressed proteins(DEPs), and KEGG analysis was carried out for these DEPs. The protein-protein interaction(PPI) networks were constructed using Cytoscape software and RT-qPCR analysis were carried out for determining the expression patterns of proteins in response to CGMMV stress. In total, we quantified 61 proteins that differentially expressed in response to CGMMV stress. These differentially expressed proteins(DEPs) included ribosomal proteins, PSI and PSⅡ proteins, ATP synthase, NADH dehydrogenase, HSP22.8, eukaryotic translation initiation factor, sucrose synthase and ethylene response factor. They were involved in KEGG pathways of photosynthesis, biosynthesis of scondary metabolites, and plant hormone signal transduction. PPI analysis identified 13, 32 and 43 DEPs for 48 h_0 h, 25 d_0 h and 25 d_48 h that involved in the known PPI network under CGMMV stress. RT-qPCR analysis obtained the expression patterns of SUS4, RPL22, NAD7, PSBH, defensin-like protein 5, ATPF, LSP1 and HSP22.8 at different infection stages of CGMMV. These results identify the proteins’ responsive to CGMMV infection, and lay a foundation for revealing molecular mechanism of CGMMV resistance in watermelon.