基于TMT蛋白质组学技术分析蝗虫微孢子虫感染飞蝗后的差异蛋白  

Differentially Expressed Proteins Analysis of Locusta migratoria Infected by Paranosema locustae Based on TMT Proteomics Technique

作　　者：张慧慧 康晗晔 刘惠 张金锐 霍帆 郭玮琦 叶小芳 季荣 扈鸿霞 ZHANG HuiHui;KANG HanYe;LIU Hui;ZHANG JinRui;HUO Fan;GUO WeiQi;YE XiaoFang;JI Rong;HU HongXia(College of Life Sciences,Xinjiang Normal University/International Research Center of Cross-Border Pest Management in Central Asia,China/Xinjiang Key Laboratory of Special Species Conservation and Regulatory Biology,Urumqi 830017;Tacheng,Research Field(Migratory Biology),Observation and Research Station of Xinjiang,Tacheng 834700,Xinjiang)

机构地区：[1]新疆师范大学生命科学学院/中亚区域跨境有害生物联合控制国际研究中心/新疆特殊环境物种多样性应用与调控重点实验室,乌鲁木齐830017 [2]塔城昆虫迁飞生物学新疆野外科学观测研究站,新疆塔城834700

出　　处：《中国农业科学》2024年第24期4884-4893,共10页Scientia Agricultura Sinica

基　　金：自治区自然科学基金面上项目(2023D01A45)。

摘　　要：【目的】通过串联质谱标签(tandem mass tag,TMT)技术测定飞蝗(Locusta migratoria)感染蝗虫微孢子虫(Paranosema locustae)前后蛋白质组变化,筛选差异表达的免疫和代谢相关蛋白,探索蝗虫微孢子虫的致病机制,为今后更好地利用蝗虫微孢子虫治蝗提供科学依据。【方法】采用实验室孵化获得的健康蝗蝻,接种5μL 1×10^(6)孢子/mL蝗虫微孢子虫。未感染的蝗蝻作为对照组与感染组置于相同条件饲养。取飞蝗血淋巴作为样本,利用TMT技术对感染组和对照组的飞蝗血淋巴进行定量蛋白质组学分析,鉴定差异蛋白。通过Gene Ontology(GO)方法,对差异蛋白的生物过程、分子功能和细胞组成进行分析。通过Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway网站对差异蛋白代谢通路进行注释。【结果】感染组和对照组之间共有128个蛋白的丰度存在显著差异。与对照组相比,66个蛋白在感染组上调,62个蛋白在感染组下调。GO分析结果显示,差异蛋白主要参与代谢过程等,且主要分布于细胞。KEGG分析结果显示,有16种蛋白在5条通路上显著富集。在免疫相关蛋白中,6个谷胱甘肽-S-转移酶(glutathione S-transferase,GST)、1个超氧化物歧化酶(superoxide dismutase,SOD)、4个热休克蛋白(heat shock protein,HSP)及2个过氧化物酶(peroxide dismutase,POD)发生了明显含量变化。在代谢相关蛋白中,6个糖代谢相关蛋白、2个氨基酸代谢相关蛋白和1个脂代谢相关蛋白的水平发生改变。【结论】飞蝗感染微孢子虫前后蛋白质组差异显著。筛选出具有不同功能的多个差异表达蛋白,主要分布于细胞。筛选出免疫相关的谷胱甘肽S-转移酶、热休克蛋白、超氧化物歧化酶和过氧化物酶等蛋白显著上调,表明免疫和刺激应答相关蛋白质在蝗虫宿主体内的免疫防御中发挥了重要作用。代谢相关蛋白的显著性提高暗示蝗虫微孢子虫感染促进宿主代谢,为其�【Objective】The objective of this study is to determine differentially expressed proteins in Locusta migratoria before and after Paranosema locustae infection by using tandem mass tag (TMT) quantitative proteomics techniques,screen differentially expressed immune and metabolic related proteins,and to explore the pathogenic mechanism of P.locustae,so as to provide a scientific basis for better use of P.locustae to control locusts in the future.【Method】The healthy nymphs obtained by laboratory incubation were inoculated with 5μL of 1×10^(6) spores/mL P.locustae.The uninfected nymphs were used as the control group and fed under the same conditions as the infected group.The hemolymph of L.migratoria was taken as a sample.TMT technique was used to analyze the quantitative proteomics of the L.migratoria hemolymph in the infected group and the control group,and the differential proteins were identified.The biological process,molecular function and cellular component of differential proteins were analyzed by the Gene Ontology (GO) method.The differential proteins metabolic pathways were annotated by the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway website.【Result】A total of 128 proteins were significantly different in abundance between the infected group and the control group,of which 66 proteins were up-regulated and 62 proteins were down-regulated in the infected group.GO analysis showed that the differential proteins were mainly involved in metabolic processes and were mainly distributed in cells.KEGG analysis showed that 16 proteins were significantly enriched in five pathways.Immune-related proteins,including six glutathione S-transferases (GSTs),one superoxide dismutase (SOD),four heat shock proteins (HSPs) and two peroxide dismutases (PODs),were significantly changed.Besides,metabolism-related proteins,including six glycometabolism-related proteins,two amino acids and one lipid metabolism-related proteins,were significantly changed.【Conclusion】There were significant differences in the pr

关 键 词：蝗虫微孢子虫 飞蝗 血淋巴 蛋白质组学 免疫相关蛋白 代谢相关蛋白 

分 类 号：S433.2[农业科学—农业昆虫与害虫防治]