中药材2024，Vol.47Issue(5)：1079-1087,9.DOI:10.13863/j.issn1001-4454.2024.05.002
广藿香青枯病菌Kosakonia cowanii Pa82致病相关基因vgrG的功能鉴定
Functional Identification of the Pathogenicity-Related Gene vgrG in Kosakonia cowanii Pa82 Causing Bacterial Wilt on Pogostemon cablin
摘要
Abstract
Objective:To identify the function of the pathogenicity-related gene vgrG in Kosakonia cowanii Pa82(wild strain Pa82)causing bacterial wilt on Pogostemon cablin.Methods:The high throughput transcriptome sequencing was performed on wild strain Pa82 and its low virulence vgrG gene mutant strain,and differential expression genes associated with vgrG gene were screened and verified by RT-qPCR.Complementation strains of vgrG gene mutant strain were constructed and their virulence phenotypes were tested.Results:From the transcriptome sequencing data of vgrG gene mutant strain,73 genes with differential expression from wild strain Pa82 were screened,and the expression levels of vgrG gene and its downstream genes JMT66_10485(10485),JMT66_10490(10490),JMT66_10500(10500)were down-regulated simultaneously.Bioinformatics analysis showed that the protein encoded by genes vgrG,10485 and 10490 were related to the assembly and function of type Ⅵ secretion system(T6SS),and the protein encoded by gene 10500 was a hypothetical protein.RT-qPCR showed that the expression trend of the four genes was consistent with that of transcriptome sequencing.Complemen-ted strains of gene vgr G,10485,10490 and 10500(C-vgrG,C-10485,C-10490,C-10500)of vgrG mutant strain were constructed by tri-parental mating method,respectively.The results of RT-qPCR showed that the expressions of vgrG,10485,10490 and 10500 genes in the above four gene complement strains were restored,and the vgrG gene complement in the vgrG gene mutant strain could restore the expressions of vgrG gene and its three downstream associated genes at the same time.Pathogenicity test showed that the four comple-mented strains restored the wild-type phenotype and interbacterial competition assay showed that complemented strains C-vgrG and C-10500 restored the wild-type phenotype.Conclusion:vgrG Gene and its downstream genes 10485,10490 and 10500 are pathogenicity-re-lated genes in Kosakonia cowanii Pa82 causing bacterial wilt disease on Pogostemon cablin,among which vgrG and 10500 genes play an important role in the antagonism between pathogenic bacteria and other bacteria.vgrG Gene can affect the pathogenicity of pathogens to Pogostemon cablin by regulating the expressions of 10485,10490 and 10500 genes downstream.关键词
广藿香/青枯病/科萨克氏菌株Pa82/vgrG基因/致病机制Key words
Pogostemon cablin(Blanco)Benth./Bacterial wilt/Kosakonia cowanii Pa82/vgrG Gene/Functional identification分类
中医学引用本文复制引用
黄德锐,张泳,王邦伟,吕丽琪,贺红..广藿香青枯病菌Kosakonia cowanii Pa82致病相关基因vgrG的功能鉴定[J].中药材,2024,47(5):1079-1087,9.基金项目
国家自然科学基金面上项目(81373901) （81373901）
广东省自然科学基金面上项目(2019A1515012151) （2019A1515012151）
