基于磷酸化组学分析四倍体香圆对干旱胁迫的响应  

Response of tetraploid Citrus wilsonii Tanaka to drought stress by phosphoproteomics analysis

作　　者：邓茜茜 任可心 魏丽娜 王浩田 胡佳 蒋景龙[1,2,3] DENG Xixi;REN Kexin;WEI Lina;WANG Haotian;HU Jia;JIANG Jinglong(School of Biological Sciences and Engineering,Shaanxi University of Technology,Hanzhong,Shaanxi 723000,China;Shaanxi Key Laboratory of Bio-resources,Hanzhong,Shaanxi 723000,China;Qinling-Bashan Mountains Bioresources Comprehensive Development Collaborative Innovation Center,Hanzhong,Shaanxi 723000,China)

机构地区：[1]陕西理工大学生物科学与工程学院,陕西汉中723000 [2]陕西省资源生物重点实验室,陕西汉中723000 [3]陕南秦巴山区生物资源综合开发协同创新中心,陕西汉中723000

出　　处：《西北植物学报》2024年第10期1567-1576,共10页Acta Botanica Boreali-Occidentalia Sinica

基　　金：陕西省重点研发计划项目(2022NY-141);陕西省教育厅项目(21JY006);秦巴生物资源与生态环境重点实验室(培育)“市校共建”科研专项(SXC-2305)。

摘　　要：【目的】基于磷酸化组学分析干旱胁迫下四倍体香圆叶片蛋白质磷酸化水平的表达规律,以期在分子水平揭示香圆对干旱的响应机制,为后续改良耐旱柑橘砧木品种提供理论依据。【方法】用IMAC亲和富集及TMT标记结合技术鉴定和分析四倍体香圆干旱胁迫后的磷酸化蛋白质,并进行功能注释和代谢通路分析。【结果】(1)共有3794个磷酸化位点和1521个磷酸化蛋白质被定量,变化倍数超过1.3倍的磷酸化蛋白质有662个(αFC>1.3),主要定位在细胞核(46.07%)与叶绿体(24.62%)中;(2)差异表达的磷酸化蛋白质主要通过结合RNA和Ca2+发挥功能;同时也参与RNA剪接、光合作用、囊泡运输中的SNARE相互作用等代谢通路;(3)92.86%差异表达的磷酸化蛋白质的基因在转录水平和蛋白水平表达变化趋势一致,其Pearson相关系数为0.893。【结论】四倍体香圆通过磷酸化修饰RNA剪接和光合作用等通路中的蛋白质来响应干旱胁迫。[Objective]Using phosphoproteomics,expression patterns of the phosphorylated proteins were analyzed with tetraploid Citrus wilsonii leaves under drought stress,aiming to reveal the mechanism of tetraploid C.wilsonii in response to drought stress and provide support for the improvement of droughttolerant citrus rootstock varieties.[Methods]Phosphorylated proteins in the leaves of tetraploid C.wilsonii after drought stress were identified and analyzed using IMAC affinity enrichment and TMT labeling technology.Functional annotation and metabolic pathway analysis were performed for the differentially expressed phosphorylated proteins.[Results](1)A total of 3794 phosphorylation sites and 1521 phosphorylated proteins were quantified.There were 662 phosphorylated proteins with a fold change exceeding 1.3(αFC>1.3),which were mainly located in the nucleus(46.07%)and chloroplasts(24.62%).(2)The differentially expressed phosphorylated proteins were mainly involved in binding RNA and Ca2+,and par-ticipating in metabolic pathways such as RNA splicing,photosynthesis,and SNARE interaction in vesicle transport.(3)RT-qPCR results showed that 92.86%of the genes coding the differentially expressed phosphorylated proteins showed similar trend of change in transcriptional and protein levels with the Pear-son correlation coefficient of 0.893.[Conclusion]Tetraploid C.wilsonii regulates the proteins involved in RNA splicing and photosynthesis pathway through phosphorylation in response to drought stress.

关 键 词：干旱胁迫 四倍体 香圆 蛋白质磷酸化 

分 类 号：Q946.1[生物学—植物学]