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生态学杂志 ›› 2020, Vol. 39 ›› Issue (8): 2792-2801.

• 技术与方法 • 上一篇    下一篇

基于iTRAQ技术筛选水稻根尖甲基汞胁迫响应差异蛋白及其生物信息学分析

龙水庭1,2,韦艳1,徐晓航2,刘琳2,仇广乐2*   

  1. (1贵州医科大学公共卫生学院, 环境污染与疾病监控教育部重点实验室, 贵阳 550025; 2中国科学院地球化学研究所环境地球化学国家重点实验室, 贵阳 550081)
  • 出版日期:2020-08-10 发布日期:2021-02-10

Screening differentially expressed proteins in root tips in response to methylmercury stress by iTRAQ technique and their bioinformatics analysis.

LONG Shui-ting1,2, WEI Yan1, XU Xiao-hang2, LIU Lin2, QIU Guang-le2*#br#   

  1. (1School of Public Health, Key Laboratory of Environmental Pollution Monitoring and Disease Control, Ministry of Education, Guizhou Medical University, Guiyang 550025, China; 2State Key Laboratory of Environmental Geochemistry, Institute of Geochemistry, Chinese Academy of Sciences, Guiyang 550081, China).
  • Online:2020-08-10 Published:2021-02-10

摘要: 为了解水稻(Oryza sativa L.)对甲基汞胁迫响应的分子机制,采用两优302为实验材料,利用同位素相对标记与绝对定量技术(iTRAQ),联合液相色谱串联质谱技术(LC-MS/MS),筛选水稻甲基汞胁迫下的根尖蛋白组差异表达蛋白,并结合生物信息学对差异蛋白进行分析。结果表明,对照组与处理组共定量了3508个蛋白质,在差异倍数(fold change)≥1.20或≤0.83,且P<0.05条件下,筛选出88个差异蛋白,其中32个蛋白表达上调,56个蛋白表达下调,其中15个差异蛋白(包括类萌发素蛋白和脂氧合酶等12个已知蛋白和3个未知蛋白)具有与金属离子结合相关属性。基因本体(gene ontology, GO)分子功能提示,差异性蛋白主要涉及催化活性、结合和转运活性等生物学过程,京都基因与基因组百科全书(Kyoto Encyclopedia of Genes and Genomes, KEGG)通路分析显示,差异蛋白显著富集于内质网蛋白加工、氧化磷酸化、淀粉与蔗糖代谢和苯丙素生物合成等代谢通路。研究结果为今后调控水稻中MeHg的吸收提供依据。

关键词: 城市河流调查, 生境质量指数, 河段尺度, 太湖流域, iTRAQ, 甲基汞胁迫, 差异蛋白, 水稻

Abstract: To understand the molecular mechanism of rice (Oryza sativa L.) in response to methylmercury (MeHg) stress, we analyzed the expressed proteins in root tips of rice Liangyou 302 under MeHg stress using isobaric tags for relative and absolute quantitation (iTRAQ) and liquid chromatographytandem mass spectrometry (LC-MS/MS). Differential proteins were elucidated in view of bioinformatics and molecular biology. The results showed that a total of 3508 proteins were quantified in both the control and MeHg treatment. According to fold change (≥1.20 or ≤0.83) andP<0.05, a total of 88 differentially expressed proteins were identified, out of which 32 proteins were upregulated and 56 proteins were down-regulated. There were 15 differentiallyexpressed proteins associated with the characteristic of metal ion binding, including 12 knownproteins of germination like proteins, lipoxygenase, and three unknown proteins. Results from the molecular functions of gene ontology (GO) suggested that differentially expressed proteins were mainly involved in catalytic activity, binding and transport activity, while the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway exhibited that they were significantly enriched in protein processing in endoplasmic reticulum, oxidative phosphorylation, starch and sucrose metabolism, and phenylpropanoid biosynthesis. Our results provide insight into regulating absorption of MeHg in rice.

Key words: urban river survey, stretch scale, stretch habitat quality index, Taihu Lake Basin, iTRAQ, methylmercury stress, differentially expressed protein, rice.