南京农业大学学报2025，Vol.48Issue(4)：848-858,11.DOI:10.7685/jnau.202403043
黄瓜绿斑驳花叶病毒(CGMMV)侵染性克隆的构建及其侵染甜瓜的转录组分析
Construction of the infectious clone of cucumber green mottle mosaic virus(CGMMV)and transcriptome analysis of infected-Cucumis melon
摘要
Abstract
[Objectives]The infectious clone of cucumber green mottle mosaic virus(CGMMV)was constructed and the transcriptome data of CGMMV-infected Cucumis melon was analyzed using RNA-Seq technology to lay the foundation for studying the resistant mechanism of C.melon against CGMMV.[Methods]The RNA was extracted from the leaves of CGMMV-infected C.melon and reverse transcribed to the cDNA,and the 3 segments divided from the full genome of CGMMV were cloned separately using cDNA as a template,and then the infectious clone pCB-CGMMV was constructed using homologous recombination.The total RNA were respectively extracted from the CGMMV-infected and healthy C.melon leaves for RNA-Seq analysis.The data was processed using the Solexa GA pipeline and SOAP software to obtain unigenes.The functions of unigenes exceeding 350 bp were annotated using GenBank and Swissprot software,and BLAST2GO tool and WEGO software were separately used for GO annotation search and functional classification of unigenes.Inter-ProScan software was utilized for COG analysis and OmicShare Tools was used for KEGG analysis.[Results]The Cucumis sativus,Citrullus lanatus,C.melon,and Nicotiana benthamiana plants were separately inoculated with pCB-CGMMV,whose leaves all displayed obvious mosaic symptoms,and CGMMV could be detected by reverse transcription polymerase chain reaction(RT-PCR)and Western blot in the systemic leaves of all the various plants.Further transcriptome analysis of the CGMMV-infected and healthy C.melon leaves showed that 1 872 differentially expressed gene(DEG)were screened,among them 1 431 were up-regulated and 441 were down-regulated.GO,COG annotation,and KEGG pathway analysis indicated that the DEG were involved in the metabolism of lipids and carbohydrates,plant signaling,MAPK cascade reaction and plant-pathogen interaction.The differential expression of nine genes were tested using quantitative RT-PCR(RT-qPCR),and these results were basically identical with the transcriptome data.[Conclusions]An infectious CGMMV Anhui isolate clone was successfully constructed,the transcriptome data of the infected-C.melon were all analyzed,and we found that the DEG of C.melon were engaged in multiple physiological metabolisms related to plant disease resistance.关键词
黄瓜绿斑驳花叶病毒/侵染性克隆构建/甜瓜/转录组分析Key words
cucumber green mottle mosaic virus/construction of the infectious clone/Cucumis melon/transcriptome analysis分类
农业科技引用本文复制引用
余维琪,蒙令瑶,徐凯,蒋磊,江彤..黄瓜绿斑驳花叶病毒(CGMMV)侵染性克隆的构建及其侵染甜瓜的转录组分析[J].南京农业大学学报,2025,48(4):848-858,11.基金项目
国家自然科学基金项目(32072386) （32072386）
