当阳鱼腥草花叶病病原分子鉴定

Molecular identification of mosaic disease on Houttuynia cordata Thumb. in Dangyang

摘要

明确当阳鱼腥草花叶病致病菌种类，为当阳鱼腥草花叶病的田间防治提供依据。采集鱼腥草花叶叶片提取当阳鱼腥草典型病样的总RNA，利用sRNA深度测序，结合生物信息学分析初步确定病原菌，利用该病毒外壳蛋白(CP)特异性引物进行RT-PCR检测，所得序列进行BLAST比对和系统进化树构建，以进一步确定病原菌种类。利用sRNA深度测序和生物信息学初步确定鱼腥草花叶病致病菌为黄瓜花叶病毒(CMV)，特异性引物扩增得到878 bp特异性条带，经BLAST比对发现其与已发表的CMV序列高度同源，相似性最高达97.84%。通过CMV病毒 CP基因的核苷酸序列构建系统进化树可得鱼腥草花叶病病毒属于CMV亚组II。引起鱼腥草花叶病的病原属于黄瓜花叶病毒II亚组，研究结果为鱼腥草花叶病防治提供了理论基础。

Abstract

The identification of pathogenic bacteria species causing Houttuynia cordata mosaic disease in Dangyang will provide a theory basis for the field control of the disease. The mosaic leaves of Houttuynia cordata were collected to extract the total RNA of typical mosaic disease samples, and sRNA deep sequencing was conducted. Based on bioinformatics analysis, the pathogen was initially identified. Using the virus coat protein (CP) specific primers for RT-PCR detection, the obtained sequences were subjected to BLAST alignment and a phylogenetic tree construction to further identify the pathogen. Using sRNA deep sequencing and bioinformatics analysis, it was initially determined that the causative virus of Houttuynia cordata mosaic disease was cucumber mosaic virus (CMV). Specific primers amplified a specific band of 878 bp, which was found to be highly homologous to published CMV sequences through BLAST alignment, with a similarity of 97.84%. By constructing a phylogenetic tree based on the nucleotide sequence of CMV virus CP gene, it can be concluded that Houttuynia mosaic virus belongs to subgroup II of CMV. This result provides a theoretical basis for the prevention and treatment of Houttuynia cordata mosaic disease.

关键词

鱼腥草 / 黄瓜花叶病毒 / sRNA深度测序 / RT-PCR

Key words

Houttuynia cordata / Cucumber mosaic virus / sRNA sequencing / RT-PCR

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基金资助

湖北省农业科技创新中心项目(2021-620-000-001-007)

国家特色蔬菜产业技术体系(CARS-24-G-17)