文献类型：期刊文献

中文题名：基于甘芪纯系群体高抗麻口病特性关联的蛋白质的筛选

英文题名：Screening of Proteins Associated with High Resistance to Makou Disease in Ganqi Pure Line Population

作者：吴永莉[1];郭苏杭[1];何春雨[1];张蕊菊[1];周珊[1];张延红[1];郭清毅[1]

第一作者：吴永莉

机构：[1]甘肃中医药大学药学院/天然药用种质资源挖掘与新品种选育协同创新中心/甘肃中医药大学杏林百草园,兰州730000

第一机构：甘肃中医药大学药学院（西北中藏药协同创新中心办公室）

年份：2025

卷号：34

期号：10

起止页码：1856

中文期刊名：西北农业学报

外文期刊名：Acta Agriculturae Boreali-occidentalis Sinica

收录：;北大核心:【北大核心2023】；

基金：甘肃省高等学校青年博士基金(2021QB-073);引进人才科研启动基金(2019YJRC-06);民生科技专项(20CX9NA070);中央引导地方科技发展专项(22ZY1QA003/2025ZY036/YDZX20206200002673);甘肃省科技计划(重点研发计划)(24YANA007/23YFFA0027/24CXKA009/22CX8NA082/25JRRA1178);高校产业支撑计划(2024CYZC-40/2020C-09)。

语种：中文

中文关键词：甘芪纯系群体;麻口病;蛋白质组学;差异表达蛋白;SDS-聚丙烯酰氨凝胶电泳

外文关键词：Ganqi pure line population;Makou disease;Proteomics;Differentially expressed pro-teins;SDS-PAGE

摘要：为探讨甘芪品系高抗麻口病特性在蛋白质水平上的抗病机制,利用iBAQ(intensity-basedabsolute-protein-quantification)非标记定量蛋白质组学技术分析鉴定具有高抗麻口病特性的甘芪纯系群体(HQ_(1)、HQ_(6)、HQ_(19))与高感麻口病黄芪(CK)在蛋白质水平上的表达差异,收集HQ_(1)、HQ_(6)、HQ_(19)及CK中的总蛋白,通过SDS-聚丙烯酰氨凝胶电泳对总蛋白进行分离,胶内酶解,提出肽段,运用LC-MS/MS获得质谱图,于Uniport蛋白数据库中检索以及iBAQ非标定量分析,筛选出差异蛋白与特异性表达蛋白,对其进行基因本体(gene ontology,GO)功能和京都基因与基因组百科全书(Kyoto encyclopedia of genes and genomes,KEGG)通路富集分析。结果表明,通过LC-MS/MS分析共鉴定到蛋白质169个,其中8个蛋白质在甘芪品系与CK之间的差异具有统计学意义(P<0.05),有6个蛋白质特异性表达,NADH脱氢酶亚基7与蔗糖合酶在差异蛋白与特异性蛋白中均存在。生物学分析发现差异蛋白大多富集在分子功能方面的ATP合成上,特异性蛋白多富集在生物过程方面,包括参与肉桂酸、麦角甾醇等生物合成以及蔗糖、乙酰辅酶A、L-苯丙氨酸分解等代谢过程,对以上蛋白进行KEGG通路富集,发现50%的蛋白参与了新陈代谢通路,包括能量代谢、氨基酸代谢、脂质代谢等。该方法筛选出了与甘芪品系高抗麻口病特性相关的多种蛋白质,包括NADH脱氢酶亚基7、蔗糖合酶、查尔酮合酶及非特异性丝氨酸/苏氨酸蛋白激酶等。
This study used intensity-based absolute quantification(iBAQ),a label-free quantitative proteomics technique,to analyze protein expression differences between Ganqi pure line strains(HQ_(1),HQ_(6),HQ_(19))exhibiting high resistance to Makou disease and a susceptible Astragalus control(CK).The aim was to elucidate the disease-resistance mechanism of Ganqi lines at the proteomic level.Total proteins were extracted from HQ_(1),HQ_(6),HQ_(19),and CK,separated via SDS-polyacrylamide gel elec-trophoresis(SDS-PAGE),digested in-gel,and analyzed using liquid chromatography-tandem mass spectrometry(LC-MS/MS).Resulting spectra were searched against the Uniprot database,and iBAQ was used for quantitative analysis.Differentially expressed and specifically expressed proteins were identified and subjected to Gene Ontology(GO)functional annotation and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis.A total of 169 proteins were identified,eight of which exhibited statistically significant differential expression(P<0.05)between Ganqi strains and CK.Six proteins were uniquely expressed in Ganqi strains.Notably,NADH dehydrogenase subunit 7 and sucrose synthase were identified among both differentially and specifically expressed proteins.GO analysis revealed that most differentially expressed proteins were enriched in molecular functions related to ATP synthesis.Specifically expressed proteins were predominantly associated with biological processes such as the biosynthesis of cinnamic acid and ergosterol,as well as the catabolism of sucrose,acetyl-CoA,and L-phenylalanine.KEGG enrichment showed that approximately 50%of these proteins participated in key metabolic pathways,including energy metabolism,amino acid metab-olism,and lipid metabolism.This study successfully identified multiple proteins associated with high resistance to Makou disease in Ganqi strains,including NADH dehydrogenase subunit 7,sucrose syn-thase,chalcone synthase,and non-specific serine/threonine protein kinases.These findings provide a foundation for analyzing the mechanism and causes of Astragalus’s resistance to Makou disease.