基于RNA-seq的香蕉枯萎病菌Cat1基因敲除突变体分析  

Analysis of Catl Knockout Mutants of Fusarium oxysporum f.sp.cubense Based on RNA-Seq

作　　者：陈亚 郑玉华 彭明[2] 李春强[2] Chen Ya;Zheng Yuhua;Peng Ming;Li Chunqiang(School of Life Science and Pharmacy,Hainan University,Haikou,570100;Institute of Tropical Biotechnology,Chinese Academy of Tropical Agricultural Sciences,Haikou,570100)

机构地区：[1]海南大学生命科学与药学院,海口570100 [2]中国热带农业科学院热带生物技术研究所,海口570100

出　　处：《分子植物育种》2023年第7期2161-2173,共13页Molecular Plant Breeding

基　　金：海南省高层次人才项目(2019RC299);国家重点研发计划项目(2018YFD1000307);中国热带农业科学院热带生物技术研究所基本科研业务费项目(1630052019028)共同资助。

摘　　要：本研究通过RNA-seq技术对Δcat1突变体菌株和Foc4野生型菌株的转录组进行了比较分析。为探明Cat1基因敲除后对香蕉枯萎病菌转录组的影响。预测到新转录本2354个,其中能够预测到基因功能的有1095个。Cat1基因敲除后,敲除突变体的基因表达相较于野生型菌株发生很大变化。差异表达基因GO功能富集发现,差异基因表达主要体现在蛋白复合物、细胞质组分、细胞蛋白代谢过程、ATP结合和转运活性等相关方面。KEGG分析发现,敲除突变体与野生型菌株的差异表达基因主要集中在核糖体合成、RNA转运,香蕉处理后差异表达基因集中在抗生素生物合成、次生代谢物生物合成等通路。Cat1基因敲除后,一些能量转运酶、加氧酶、氨基酸转移酶活动加强,一些激酶、氧化酶、蛋白酶受香蕉诱导活性增加。Cat1的敲除导致多种糖转运蛋白的大幅下调表达,且大大影响锌脂蛋白和细胞周期蛋白等表达,从而对菌株生命活动和致病力产生影响。Cat1敲除后,其他过氧化氢酶类的表达量补偿性增加,但催化生成过氧化氢的SOD表达也增加,可能是敲除突变体致病力减弱的原因之一。本研究为进一步揭示香蕉枯萎病菌的致病机理提供理论依据。In this study,the transcriptomes of Catl mutant strains and Foc4 wild-type strains were comparatively analyzed by RNA-seq technology.To explore the effect of Cat1 knockout on the transcriptome of Foc.2354 new transcripts were predicted,of which 1095 gene functions could be predicted.After the Catl gene was knocked out,the gene expression of the knockout mutant changed greatly compared with the wild-type strain.GO function enrichment of differentially expressed genes found that differential gene expression was mainly reflected in protein complexes,cytoplasmic components,cellular protein metabolism,ATP binding and transport activities.KEGG analysis found that the differentially expressed genes between the knockout mutant and the wild-type strain were mainly concentrated in ribosome synthesis and RNA transport.The differentially expressed genes after banana treatment were concentrated in the pathways of antibiotic biosynthesis,secondary metabolite biosynthesis and so on.After the Catl gene was knocked out,the activities of some energy transferases,oxygenases,and amino acid transferases were enhanced,and the activities of some kinases,oxidases,and proteases were increased by banana-induced activities.The knockout of Catl lead to a significant down-regulation of the expression of various sugar transporters,and greatly affected the expression of zinc finger protein and cyclins,thereby affecting the life activity and pathogenicity of the strain.After the Cat l gene was knocked out,the expression levels of other catalase enzymes increased compensatively,but the expression of SOD,which catalyzes the production of hydrogen peroxide,also increased,which may be one of the reasons for the weakened pathogenicity of the knockout mutants.This study lays the foundation for further revealing the pathogenic mechanism of Foc.

关 键 词：过氧化氢酶 敲除突变体 转录组测序 尖孢镰刀菌古巴专化型 基因表达 

分 类 号：S436.68[农业科学—农业昆虫与害虫防治]