基于全基因组关联分析的香蕉枯萎病菌致病基因挖掘与功能研究

Mining and functional study of pathogenic genes of Fusarium oxysporum f. sp. cubense based on genome-wide association analysis

  • 摘要: 【目的】深入挖掘香蕉枯萎病菌致病相关基因,并解析其基因特性,为香蕉抗枯萎病品种选育及新药物靶标的开发打下基础。【方法】基于遗传背景将供试299株香蕉枯萎病菌的致病力分别依据数量性状和质量性状进行分组,使用混合线性模型的Gemma软件,运用全基因组关联分析(GWAS)技术对香蕉枯萎病菌致病力表型与全基因组重测序数据进行关联分析;进一步采用ProtParam、SingleIP和SAMRT等生物信息学软件解析候选基因的理化性质及初级结构。【结果】运用GWAS共关联到151个与致病力相关的单核苷酸多态性(SNP)位点。初步确定3个不同类型基因FocScpFocChpFocGhf12与枯萎病菌致病力密切相关,进一步对FocScpFocChpFocGhf12基因的编码蛋白进行生物信息学分析并预测其功能,结果显示,FocScp基因的cDNA编码区全长948 bp,编码314个氨基酸残基,大小约33.27 kD,含有N-端信号肽,预测为胞外分泌蛋白;FocChp基因的cDNA编码区全长1302 bp,编码433个氨基酸残基,大小约49.17 kD,亚细胞定位在细胞核,预测为含有3个锌指结构域的转录因子;FocGf12基因的cDNA编码区全长1533 bp,编码480个氨基酸残基,大小约53.54 kD,该蛋白存在跨膜结构域,具有GPI修饰位点,亚细胞定位在胞外,预测为糖苷水解酶家族12蛋白。【结论】通过GWAS获得香蕉枯萎病菌3个致病相关基因FocScpFocChpFocGhf12,其编码蛋白分别为分泌蛋白、转录因子和结构蛋白。

     

    Abstract: 【Objective】In-depth exploration of the pathogenicity related genes in Fusarium oxysporum f. sp. cubense, and analysis of their genetic characteristics were conducted to laid foundation for the breeding of banana varieties resistant to fusarium wilt,as well as the development of new drug targets.【Method】Based on the genetic backgrounds,the pathogenicity of 299 tested strains of F. oxysporum f. sp. cubenseF. oxysporum f. sp. cubense and the genomewide resequencing data. Additionally,ProtParam,SingleIP and SAMRT online software tools were utilized to analyze the physicochemical properties and primary structure of candidate genes.【Result】Using GWAS,a total of 151 pathogenicity related single nucleotide polymorphism(SNP)loci were associated. It was preliminarily determined that 3 different types of genes FocScpFocChpFocGhf12 were closely related to the pathogenicity of F. oxysporum f. sp. cubense. Further bioinformatics analysis was performed on the encoded proteins of FocScpFocChpFocGhf12 genes and their functions were predicted. The results revealed that the full-length cDNA coding region of FocScp gene was 948 bp,and encoded 314 amino acid residues with size of about 33.27 kD and contained an N-terminal signal peptide,which was predicted to be an extracellular secretory protein. The full-length cDNA coding region of FocChp gene was 1302 bp,encoding 433 amino acid residues with size of about 49.17 kD,with subcellular localization in the nucleus,and was predicted to be a transcription factor containing 3 zinc finger domains. The full-length cDNA coding region of FocGhf12 gene was 1533 bp,encoding 480 amino acid residues with size of about 53.54 kD. The protein had transmembrane domain, GPI modification sites,and subcellular localization outside the cell. It was predicted to be a glycoside hydrolase family 12 protein.【Conclusion】Three pathogenicity related genes(FocScpFocChpFocGhf12)of F. oxysporum f. sp. cubense are obtained by GWAS,and their encodesd proteins are secretory protein,transcription factor and structural protein.

     

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