浙江菜豆CPMMV变异株的鉴定及其TGB蛋白定位  

Identification of Cowpea Mild Mottle Virus Variants in Zhejiang and Localization of TGB Proteins

作　　者：张子鑫 陈淑婷 周伟欣 张意涵 鲁倩 李中义 吴昕扬 徐沛 ZHANG Zixin;CHEN Shuting;ZHOU Weixin;ZHANG Yihan;LU Qian;LI Zhongyi;WU Xinyang;and XU Pei(Key Laboratory of Specialty Agri-Product Quality and Hazard Controlling Technology of Zhejiang,College of Life Sciences,China Jiliang University,Hangzhou 310018,China)

机构地区：[1]中国计量大学生命科学学院,浙江省特色农产品品质及危害物控制技术重点实验室,杭州310018

出　　处：《园艺学报》2025年第1期123-135,共13页Acta Horticulturae Sinica

基　　金：国家自然科学基金项目(32202470);浙江省自然科学基金项目(LY24C150006);省部共建农产品质量安全危害因子与风险防控国家重点实验室开放课题项目(2021DG700024-KF202217)。

摘　　要：2022年10—11月,在浙江省杭州市钱塘区观察到疑似病毒侵染的菜豆植株,病毒组测序和PCR技术检测出豇豆轻斑驳病毒(cowpea mild mottle virus,CPMMV)、黄瓜花叶病毒(cucumber mosaic virus,CMV)和紫云英矮缩病毒(milk vetch dwarf virus,MDV),并将CPMMV命名为CPMMV-ZJ(GenBank:OR667247)。利用病毒鉴定、全基因组序列扩增、系统发育分析及亚细胞定位等方法,明确了CPMMV-ZJ的全基因组序列特征、病毒演化及其病毒三联蛋白(triple gene block protein,TGB)的亚细胞定位。结果表明,CPMMV-ZJ的基因组全长为8226 bp,包含6个开放阅读框。3′和5′区域分别有1个147 bp和75 bp的非编码区(untranslated regions,UTR)。序列分析结果表明,CPMMV浙江分离物与CPMMV安徽分离物的全基因组序列一致性最高(98.03%),并与国内分离物共同聚类在一个小分支上,但与国外分离物(美国、印度、巴西等地)序列一致性较低(63.71%~82.49%),提示病毒变异与地理因素密切相关。对浙江分离物与江苏分离物TGB蛋白的亚细胞定位比较分析发现,二者无明显差异,但TGB2浙江分离物在叶片中的表达导致明显的叶绿体核周聚集现象,推测与植物防御反应有关。During the period from October to November 2022,an outbreak of suspected virus-infected common bean occurred in the Qiantang District of Hangzhou City,Zhejiang Province.Subsequent viral genome sequencing and PCR confirmed the presence of cowpea mild mottle virus(CPMMV),cucumber mosaic virus(CMV),milk vetch dwarf virus(MDV).CPMMV was designated as CPMMV-ZJ(GenBank:OR667247).By virus identification,genome sequence amplification,phylogenetic analysis and subcellular localization analysis,the sequence structure of CPMMV-ZJ and the localization of the triple gene block proteins(TGB)were elucidated.The genome of CPMMV-ZJ encompassed a total length of 8226 bp,consisting of six open reading frames.Additionally,both the 3′and 5′regions contained non-coding regions(UTRs)of 147 and 75 bp,respectively.Phylogenetic analysis of CPMMV-ZJ revealed the highest sequence identity(98.03%)with the CPMMV Anhui isolate,whereas lower identities(63.71%-82.49%)were observed when comparing to foreign isolates from the United States,India,and Brazil,indicating virus evolution is related to geographical factors.Subcellular localization analysis demonstrated similar localization of TGBs between the Zhejiang and Jiangsu isolates.Notably,overexpression TGB2 of the Zhejiang isolate leads to a clustering of chloroplasts around the nuclei,suggesting its potential involvement in plant defense responses.

关 键 词：菜豆 豇豆轻斑驳病毒 序列分析 亚细胞定位 叶绿体核周聚集 

分 类 号：S643.1[农业科学—蔬菜学]