基于iTRAQ-PRM技术筛选不同类型土壤烤烟根系差异表达蛋白  

Screening of differentially expressed proteins in flue-cured tobacco roots in different soil types based on iTRAQ-PRM

作　　者：杨睿 查文菊 陈颐 何承刚[2] 王涛[3] 何聪莲 费明亮 YANG Rui;ZHA Wenju;CHEN Yi;HE Chenggang;WANG Tao;HE Conglian;FEI Mingliang(Agronomy Research Center,Yunnan Academy of Tobacco Agriculture Science,Kunming 650031,Yunnan,China;College of Tobacco Science,Yunnan Agricultural University,Kunming 650201,Yunnan,China;Qujing tobacco company of Yunnan,Qujing 671000,Yunnan,China)

机构地区：[1]云南省烟草农业科学研究院,云南省昆明市650031 [2]云南农业大学,云南省昆明市650201 [3]云南省烟草公司曲靖市公司,云南省曲靖市671000

出　　处：《中国烟草学报》2022年第1期86-97,共12页Acta Tabacaria Sinica

基　　金：云南省应用基础面上项目(2018BB019);云南省烟草专卖局科技项目(2020530000241025)。

摘　　要：【目的】从蛋白质水平揭示烤烟根系对不同类型土壤的响应差异。【方法】以烤烟品种K326为材料,选取石灰岩土(沙壤土)、水稻土和红壤土,测定3种类型土壤烤烟根系活力、硝酸还原酶(NR)、谷氨酰胺合成酶(GS)和天冬氨酸合成酶(AS)活性;利用iTRAQ技术鉴定3种类型土壤烤烟根系差异表达蛋白,对所获得的差异蛋白进行生物信息学分析,从中选择16个蛋白进行平行反应监测(PRM)验证。【结果】(1)红壤土的烤烟根系活力、NR、GS和AS活性均高于沙壤土和水稻土;(2)从水稻土vs沙壤土、沙壤土vs红壤土、水稻土vs红壤土3组对比中分别鉴定出699、650、569个烤烟根系差异表达蛋白,其中上调/下调蛋白分别为412/287、291/359和323/246个;(3)差异蛋白质主要有催化、结合、转运活性等功能;(4)差异蛋白功能富集于次生代谢产物的生物合成、代谢途径和苯丙烷类生物合成等代谢途径;(5)红壤土烤烟根系与抗逆性、碳水化合物和能量代谢、蛋白质合成等相关的差异蛋白表达均高于沙壤土和水稻土。【结论】利用iTRAQ标记技术,结合PRM验证技术筛选出不同类型土壤烤烟根系差异表达蛋白,为获取响应关键蛋白奠定基础。[Background]This study aims to reveal the response of flue-cured tobacco roots to different types of soil at the protein level.[Methods]Using flue-cured tobacco variety K326 as material,the root activity,nitrate reductase(NR),glutamine synthetase(GS)and aspartate synthetase(AS)activities of flue-cured tobacco roots in three types of soil including calcareous soil(sandy soil),paddy soil and red soil were measured.iTRAQ technology was used to identify the differentially expressed proteins of flue-cured tobacco roots in three types of soil,and the obtained differential proteins were analyzed by bioinformatics.On this basis,16 proteins were selected for parallel reaction monitoring(PRM)validation.[Results](1)The root activity and activities of NR,GS,AS of flue-cured tobacco roots in red soil were higher than those in sandy soil and paddy soil;(2)699,650 and 569 differentially expressed proteins were identified from the comparison of paddy soil vs sandy soil,sandy soil vs red soil,paddy soil vs red soil,respectively.The up-regulated/down regulated proteins were 412/287,291/359 and 323/246,respectively;(3)Differentially expressed proteins are mainly involved in metabolic process,cellular process,stress response and other biological processes.They exist in cells,cell components and organelles,and have catalytic activity,binding activity and transport activity;(4)Their functions are enriched in the biosynthesis and metabolic pathways of secondary metabolites and phenylpropanoid biosynthesis;(5)The stress resistance,carbohydrate and energy metabolism,and protein synthesis of flue-cured tobacco roots in red soil were higher than those in sandy soil and paddy soil;(6)PRM analysis showed that results of iTRAQ proteomics were correct and reliable.[Conclusion]iTRAQ labeling technology was used to screen the root differentially expressed proteins of flue-cured tobacco in different types of soil combined with PRM verification technology,which provided some theoretical basis and technical support for choosing the appropriate soil

关 键 词：ITRAQ 烟草根系 土壤类型 氮代谢 蛋白质组学 

分 类 号：S572[农业科学—烟草工业]