禾谷镰刀菌醛脱氢酶(ALDH)基因敲除突变体的转录组分析  

Transcriptome analysis of aldehyde dehydrogenase (ALDH)gene knockout mutant in Fusarium graminearum

作　　者：唐磊 翟焕趁[1] 葛锦雯 张帅兵[1] 吕扬勇[1] 魏闪 马平安 胡元森[1] TANG Lei;ZHAI Huan-chen;GE Jin-wen;ZHANG Shuai-bing;LYU Yang-yong;WEI Shan;MA Ping-an;HU Yuan-sen(College of Biological Engineering,Henan University of Technology,Zhengzhou,Henan 450001,China)

机构地区：[1]河南工业大学生物工程学院,河南郑州450001

出　　处：《南方农业学报》2024年第3期733-744,共12页Journal of Southern Agriculture

基　　金：国家自然科学基金项目(31972176);河南工业大学自然科学创新基金项目(2020ZKCJ01);企业委托科技项目(H2022sw175)。

摘　　要：【目的】对野生型PH-1菌株和醛脱氢酶(ALDH)基因FGSG_04194敲除突变体(ΔFg04194)进行转录组测序,分析禾谷镰刀菌ALDH调控菌丝生长和脱氧雪腐镰刀菌烯醇(DON)生成情况,为深入探究禾谷镰刀菌ALDH的生物学功能、DON生成机制及致病机理提供了理论依据。【方法】接种禾谷镰刀菌野生型PH-1、ΔFg04194突变体和基因回补突变体(ΔFg04194-C)菌株至CM培养基和小麦基质中,分析FGSG_04194基因对菌丝生长和DON生成的影响。利用高通量转录组测序技术对野生型PH-1菌株和ΔFg04194突变菌株在CM培养基中培养72 h的菌丝进行转录组测序,对筛选出的差异表达基因(DEGs)进行GO功能注释和KEGG信号通路富集分析,利用实时荧光定量PCR对转录组测序结果进行验证。【结果】ΔFg04194突变体的菌落直径和生成的DON含量较野生型PH-1和ΔFg04194-C突变体菌株显著减少(P<0.05,下同)。野生型PH-1和ΔFg04194突变体转录组原始数据经过滤后获得34.9 Gb Clean data,共鉴定出329个DEGs,其中263个基因表达显著上调,66个基因表达显著下调。GO功能注释结果显示,DEGs显著富集在质膜成分、碳水化合物代谢和物质转运过程等条目上。KEGG信号通路富集分析显示,DEGs显著富集在碳水化合物代谢通路、氨基酸代谢通路、脂质代谢、能量代谢、生物降解代谢通路、细胞生长和死亡相关途径、信号转导途径等。将FGSG_04194基因调控下有代表性的DEGs分成五大类:几丁质合成相关的基因、跨膜转运蛋白相关的基因、转录因子相关基因、脂质代谢相关基因和能量代谢相关的基因。ΔFg04194突变体的ATP含量显著高于野生型PH-1和ΔFg04194-C突变体。运用实时荧光定量PCR检测7个DEGs的表达情况,其结果与转录组测序结果基本一致。【结论】FGSG_04194影响禾谷镰刀菌的菌丝生长和DON生成,并在ATP生成中起负调控作用,其机制可能与调控转录因子的表达、�【Objective】The purpose of the study was to perform transcriptome sequencing of wild-type PH-1 strain and aldehyde dehydrogenase(ALDH)gene FGSG_04194 knockout mutant(ΔFg04194),to analyze the regulation of mycelial growth and deoxynivalenol(DON)production by ALDH in Fusarium graminearum,and to provide a theoretical basis for in-depth study of the biological function of ALDH,DON production mechanism and pathogenesis of F.graminearum.【Method】The wild-type PH-1,ΔFg04194 mutant and gene complementation mutant(ΔFg04194-C)strains of F.graminearum were inoculated into CM medium and wheat grains,respectively.The impact of the FGSG_04194 gene on mycelial growth and DON production was analyzed.By employing high-throughput transcriptome sequencing technology,mycelium from wild-type PH-1 andΔFg04194 mutant strains cultured in CM medium for 72 h were conducted to transcriptome sequencing.The screened differentially expressed genes(DEGs)were analyzed through GO functional annotation and KEGG signaling pathway analysis,and the transcriptome sequencing results were confirmed using real-time fluorescence quantitative PCR.【Result】In comparison to the wild-type PH-1 andΔFg04194-C mutant strains,theΔFg04194 mutant exhibited significantly reduced colony diameter and generated DON content(P<0.05,the same below).The raw data of the wild-type PH-1 andΔFg04194 mutant transcriptome were filtered,resulting in 34.9 Gb of Clean data.A total of 329 DEGs were identified,with 263 genes showing significant up-regulation and 66 genes displaying significant down-regulation in expression.The GO functional annotation results indicated a significant enrichment of DEGs in categories of plasma membrane components,carbohydrate metabolism and substance transport processes.KEGG metabolic pathway analysis demonstrated a significant enrichment of DEGs in carbohydrate metabolism pathway,amino acid metabolism pathway,lipid metabolism pathway,energy metabolism pathway,biodegradation metabolism pathway,pathways associated with cell growth and d

关 键 词：禾谷镰刀菌 醛脱氢酶 转录组测序 差异表达基因 

分 类 号：S432.44[农业科学—植物病理学] S435.121.4[农业科学—农业昆虫与害虫防治]