中文摘要:双生病毒已经导致了作物生产的严重损失。为了探究寄主防御双生病毒潜在的机制,常用基于相对和绝对定量的同位素标记(iTRAQ)的定量蛋白质组学方法探索本氏烟叶片蛋白质早期响应中国番茄黄化曲叶病毒(TYLCCNV)伴随betasatellite(TYLCCNB)入侵的表现形式。本研究结果提高了众人对寄主抵抗双生病毒的蛋白质组学方面的理解, 同时也表明利用生物信息学和蛋白质组学结合VIGS分析是研究植物对双生病毒的系统反应和揭示植物-病毒交互作用的有效途径。
外文摘要:Geminiviruses have caused serious losses in crop production. To investigate the mechanisms underlying host defenses against geminiviruses, an isobaric tags for relative and absolute quantification (iTRAQ)-based quantitative proteomic approach was used to explore the expression profiles of proteins in Nicotiana benthamiana (N. benthamiana) leaves in response to tomato yellow leaf curl China virus (TYLCCNV) with its betasatellite (TYLCCNB) at an early phase. In total, 4155 proteins were identified and 272 proteins were changed differentially in response to TYLCCNV/rYLCCNB infection. Bioinformatics analysis indicated that S-adenosyl-L-methionine cycle II was the most significantly up-regulated biochemical process during TYLCCNV/TYLCCNB infection. The mRNA levels of three proteins in S-adenosyl-L-methionine cycle H were further analyzed by qPCR, each was found significantly up-regulated in TYLCCNV/TYLCCNB-infected N. benthamiana. This result suggested a strong promotion of the biosynthesis of available methyl groups during geminivirus infections. We further tested the potential role of RdDM in N. benthamiana by virus-induced gene silencing (VIGS) and found that a disruption in RdDM resulted in more severe infectious symptoms and higher accumulation of viral DNA after TYLCCNV/TYLCCNB infection. Although the precise functions of these proteins still need to be determined, our proteomic results enhance the understanding of plant antiviral mechanisms. Biological significance: One of the major limitations to crop growth in the worldwide is the prevalence of geminiviruses. They are able to infect food and cash crops and cause serious crop failures and economic losses worldwide, especially in Africa and Asia. Tomato yellow leaf curl China virus (TYLCCNV), which causes severe viral diseases in China, is a monopartite geminivirus associated with the betasatellite (TYLCCNB). However, the mechanisms underlying the TYLCCNV/TYLCCNB defense in plants are still not fully understood at the molecular level. In this study, the combined proteomic, bioinformatic and VIGS analyses revealed that TYLCCNV/rYLCCNB invasion caused complex proteomic alterations in the leaves of N. benthamiana involving the processes of stress and defense, energy production, photosynthesis, protein homeostasis, metabolism, cell structure, signal transduction, transcription, transportation, and cell growth/division. Promotion of available methyl groups via the S-adenosyl-L-methionine cycle II pathway in N. benthamiana appeared crucial for antiviral responses. These findings enhance our understanding in the proteomic aspects of host antiviral defenses against geminiviruses, and also demonstrate that the combination of proteomics with bioinformatics and VIGS analysis is an effective approach to investigate systemic plant responses to geminiviruses and to shed light on plant-virus interactions.
外文关键词:N. benthamiana; TYLCCNV/TYLCCNB; Quantitative proteomics; Defense mechanisms
作者:Zhong, Xueting; Wang, Zhan Qi; Xiao, Ruyuan; 等
作者单位:浙江大学
期刊名称:JOURNAL OF PROTEOMICS
期刊影响因子:3.867
出版年份:2017
出版刊次:1
点击下载:通过烟叶蛋白质组分析研究RNA介导的DNA甲基化(RdDM)在防御双生病毒侵染中的作用