基于4D label-free技术的水稻成熟种子蛋白质组学研究  

Proteomics study of mature rice seeds based on 4D label-free technology

作　　者：梁文轩 王月萍 陈升杰 李敏诗 韩儒 周玉亮[1] LIANG Wenxuan;WANG Yueping;CHEN Shengjie;LI Minshi;HAN Ru;ZHOU Yuliang(Guangdong Provincial Key Laboratory of Plant Molecular Breeding/College of Agriculture,South China Agricultural University,Guangzhou 510642,China)

机构地区：[1]广东省植物分子育种重点实验室/华南农业大学农学院,广东广州510642

出　　处：《华南农业大学学报》2023年第5期742-749,共8页Journal of South China Agricultural University

基　　金：国家自然科学基金面上项目(31771888);“双一流”学科建设和提升专项(2021B10564001);广东省自然科学基金面上项目(2022A1515010843)。

摘　　要：【目的】从蛋白水平揭示水稻种子成熟的分子基础,探究调控水稻种子成熟的关键蛋白和代谢通路。【方法】选用授粉后30 d的成熟水稻种子,利用4D label-free定量蛋白质组学进行质谱鉴定,通过生物信息学技术分析蛋白的亚细胞定位、结构域、GO注释和KEGG通路注释。【结果】总共鉴定了3484个种子成熟期的蛋白,相对分子质量大多在10000~100000之间,主要分布于细胞质、细胞核、叶绿体、线粒体和质膜上;结构域主要涉及蛋白质翻译的RNA识别基序和蛋白磷酸化修饰的蛋白激酶结构域;GO分析表明,成熟种子的蛋白主要参与了细胞过程和代谢过程,主要涉及催化活性和结合等功能,大多分布在细胞、细胞组分、细胞器和细胞膜等部位;KEGG分析发现,蛋白主要富集在核糖体、内质网中的蛋白质加工、氧化磷酸化和糖酵解等途径,推测蛋白质的翻译、加工和能量代谢是水稻种子成熟期的主要分子事件;进一步鉴定了脱落酸(Abscisic acid,ABA)信号和吲哚乙酸(Indoleacetic acid,IAA)代谢的相关蛋白,同时也发现了NAC(NAM、ATAF1/2和CUC2)家族的转录因子。【结论】贮藏物质的积累和能量代谢是水稻种子成熟期的典型特征,ABA和IAA信号途径参与了种子成熟过程。【Objective】To reveal the molecular basis of rice seed maturation at the protein level,and explore the key proteins and metabolic pathways involved in regulating rice seed maturation.【Method】The experiment used mature rice seeds at 30 days after pollination,and conducted mass spectrum identification using 4D label-free quantitative proteomics.Bioinformatics techniques were used to analyze the subcellular localization,domains,GO annotations,and KEGG pathway annotations of the proteins.【Result】A total of 3484 seed maturation proteins were identified,with most having a relative molecular mass between 10000 and 100000.They were mainly distributed in the cytoplasm,nucleus,chloroplasts,mitochondria,and plasma membrane.The protein domain mainly involved the RNA recognition motifs of protein translation and the protein kinase domains of protein phosphorylation modification.GO analysis showed that the proteins of mature seeds were mainly involved in cellular and metabolic processes,mainly related to catalytic activity and binding functions,and were mostly distributed in cell,cellular component,organelle,and cytomembrane.KEGG analysis revealed that proteins were mainly enriched in pathways such as ribosome,protein processing in endoplasmic reticulum,oxidative phosphorylation,and glycolysis.It was speculated that protein translation,processing,and energy metabolism were the main molecular events of rice seed maturation.Furthermore,proteins related to abscisic acid(ABA)signaling and indoleacetic acid(IAA)metabolism were identified,and transcription factors of the NAM,ATAF1/2 and CUC2(NAC)family were also discovered.【Conclusion】The accumulation of storage substances and energy metabolism are typical characteristics of rice seed maturation,and ABA and IAA signaling pathways are involved in the process of seed maturation.

关 键 词：水稻 种子成熟 代谢通路 植物激素 

分 类 号：S511[农业科学—作物学] S330