生物技术通报2026，Vol.42Issue(1)：271-278,8.DOI:10.13560/j.cnki.biotech.bull.1985.2025-0944
小西葫芦黄花叶病毒弱毒疫苗的构建及其交叉保护防治作用
Construction of a Mild Vaccine of Zucchini Yellow Mosaic Virus and Its Cross-protective Efficacy
摘要
Abstract
[Objective]By investigating the effects of the mutation at the 364th conserved lysine(K364)in helper component-proteinase(HC-Pro)on the virulence of zucchini yellow mosaic virus(ZYMV),an attenuated ZYMV mutant were developed,and its genetic stability and cross-protection efficacy were assessed.Our study provided the theoretical basis for developing sustainable control strategies against ZYMV using attenuated strains.[Method]Using site-directed mutagenesis,the K364 residue in HC-Pro was substituted with aspartic acid(D)to investigate the effect of the K364D mutation on the virulence of ZYMV and the RNA silencing suppression activity of HC-Pro,thereby constructing the attenuated ZYMV-HC-ProK364D mutant.Furthermore,the genetic stability of ZYMV-HC-ProK364D mutant was assessed through serial passage of progeny viruses.The cross-protection efficacy of ZYMV-HC-ProK364D mutant was evaluated through cross-protection assays.[Result]The K364D mutation in HC-Pro attenuated ZYMV virulence,alleviated typical yellowing symptoms and reduced ZYMV accumulation in Cucumis melo plants.Simultaneously,K364D mutation suppressed the RNA silencing suppression activity of ZYMV HC-Pro.The attenuated mutant ZYMV-HC-ProK364D-GFP maintained genetic stability without reversion mutation after three successive passages in C.melo plants.In cross-protection assays,ZYMV-HC-ProK364D mutant induced significant resistance in C.melo plants against the wild-type ZYMV infection when the protection interval was 11 d.[Conclusion]The conserved K364 residue in HC-Pro was crucial in determining the RNA silencing suppression activity and the virulence of ZYMV.The ZYMV-HC-ProK436D mutant present excellent cross-protection efficacy and is a potential mild vaccine against the wild-type ZYMV infection in C.melo plants.关键词
小西葫芦黄花叶病毒/辅助成分-蛋白酶/弱毒疫苗/遗传稳定性/交叉保护Key words
Zucchini yellow mosaic virus/helper component-proteinase/mild vaccine/genetic stability/cross-protection引用本文复制引用
张悦,任倩,栾雅梦,李翌琛,孙无瑕,任姝颖,许小洁,孙晓辉..小西葫芦黄花叶病毒弱毒疫苗的构建及其交叉保护防治作用[J].生物技术通报,2026,42(1):271-278,8.基金项目
山东省重点研发计划项目(2023LZGC002),国家自然科学基金项目(32302310),山东省自然科学基金项目(ZR2023QC094),山东省烟台市高校与地方企业合作项目(2021XDRHXMPT09) （2023LZGC002）
