侵染蝴蝶兰的病毒检测及ToMMV蝴蝶兰分离物的cp基因序列分析  

Identification of the Viruses Infecting Phalaenopsis aphrodite and cp Gene Sequence Analysis of ToMMV P.aphrodite Isolate

作　　者：曹玉莲 谢含雨 谭冠林 陈小姣 李凡[1] CAO Yulian;XIE Hanyu;TAN Guanlin;CHEN Xiaojiao;LI Fan(State Key Laboratory for Conservation and Utilization of Bio-Resources in Yunnan,Yunnan Agricultural University,Kunming 650201,China;Modern Education Technology Center,Yunnan Agricultural University,Kunming 650201,China)

机构地区：[1]云南农业大学云南生物资源保护与利用国家重点实验室,云南昆明650201 [2]云南农业大学现代教育技术中心,云南昆明650201

出　　处：《云南农业大学学报(自然科学版)》2025年第1期42-50,共9页Journal of Yunnan Agricultural University:Natural Science

基　　金：国家重点研发计划(2022YFD1400700);国家自然科学基金项目(31660039)。

摘　　要：【目的】调查云南昆明市售蝴蝶兰的病毒病发生情况。【方法】利用建兰花叶病毒(cymbidium mosaic virus,CymMV)、齿兰环斑病毒(odontoglossum ringspot virus,ORSV)、番茄斑驳花叶病毒(tomato mottle mosaic virus,ToMMV)等病毒特异性引物对收集自昆明花卉市场的17份蝴蝶兰样品进行RT-PCR检测,并对扩增获得的相关病毒基因片段进行序列比对。【结果】在7份蝴蝶兰样品中检出ORSV、CymMV、番茄斑萎病毒(tomato spotted wilt orthotospovirus,TSWV)和ToMMV,检出率分别为35.29%、23.53%、5.88%和5.88%。其中,ORSV和CymMV为当前昆明市售蝴蝶兰的主要病毒,且2种病毒的复合侵染率为23.53%;ToMMV为首次在蝴蝶兰中检出。未检测到黄瓜花叶病毒(cucumber mosaic virus,CMV)、凤仙花坏死斑病毒(impatiens necrotic spot orthotospovirus,INSV)和番茄环纹斑点病毒(tomato zonate spot orthotospovirus,TZSV)。蝴蝶兰为新发现的ToMMV自然新寄主,人工接种条件下感染ToMMV的蝴蝶兰植株无明显症状。对获得的ToMMV蝴蝶兰分离物完整cp基因序列(GenBank登录号:PP445225)与国内外已报道的19个ToMMV分离物cp基因核苷酸序列进行比对,发现这20个ToMMV分离物的cp基因核苷酸序列一致性为98.12%~99.79%。通过构建ToMMV cp基因系统进化树,发现ToMMV蝴蝶兰分离物与ToMMV云南辣椒分离物、云南豌豆分离物的亲缘关系最近,与国外分离物亲缘关系较远。【结论】ORSV和CymMV为昆明市售蝴蝶兰携带的优势病毒。首次报道了ToMMV在蝴蝶兰及兰科植物中的侵染,携带有ToMMV的无症状蝴蝶兰可能造成病毒的远距离传播。[Purpose]To investigate the occurrence of viral disease on Phalaenopsis aphrodite sold in the market in Kunming,Yunnan Province.[Methods]RT-PCR detection was conducted in 17 P.aphrodite samples collected from the market with virus-specific primers including cymbidium mosaic virus(CymMV),odontoglossum ringspot virus(ORSV),tomato mottle mosaic virus(ToMMV),and so on.The amplified fragments of related viral genes were cloned,sequenced and comparative analyzed.[Results]Four viruses were detected in seven P.aphrodite samples:ORSV,CymMV,tomato spotted wilt orthotospovirus(TSWV)and ToMMV,with detection rates of 35.29%,23.53%,5.88%and 5.88%,respectively.Among them,ORSV and CymMV were the two dominant viruses of P.aphrodite in Kunming market,and the co-infection rate of them was 23.53%.Meanwhile,ToMMV was detected in P.aphrodite for the first time.No cucumber mosaic virus(CMV),impatiens necrotic spot orthotospovirus(INSV)and tomato zonate spot orthotospovirus(TZSV)was detected.P.aphrodite is a new natural host for ToMMV,and the ToMMV inoculated P.aphrodite plants showed no obvious symptoms.Nucleotide sequence identity of the complete cp gene sequence of ToMMV P.aphrodite isolate(YKMHDL,GenBank Acc.No.:PP445225)was analyzed with those of 19 ToMMV isolates worldwide.The cp gene nucleotide sequence identity of these 20 ToMMV isolates was from 98.12%to 99.79%.Phylogenetic tree based on the cp gene of different ToMMV isolates revealed that the ToMMV-YKMHDL isolate was closely related to the ToMMV Yunnan pepper isolate and Yunnan pea isolate,but it was more distantly related to the isolates outside China.[Conclusion]ORSV and CymMV are the two dominant viruses found in P.aphrodite sold in Kunming.This is the first report of natural infection of ToMMV in P.aphrodite as well as plants in family Orchidaceae.The asymptomatic presence of ToMMV in P.aphrodite has the potential to lead to extensive and long-distance transmission of the virus.

关 键 词：蝴蝶兰 病毒鉴定 番茄斑驳花叶病毒(ToMMV) CP基因 系统进化 

分 类 号：S682.301[农业科学—观赏园艺]