广西罗汉果病毒病原调查及两种病毒的分子特征  

Investigation of viruses in Siraitia grosvenorii in Guangxi and the molecular characterization of two viruses

作　　者：黄昭戟 周琼[1] 李刚[2] 唐美琼[2] 裴艳艳 覃犇 胡营[2] 张占江[2] HUANG Zhaoji;ZHOU Qiong;LI Gang;TANG Meiqiong;PEI Yanyan;QIN Ben;HU Ying;ZHANG Zhanjiang(College of Agriculture,Guangxi University,Nanning 530004,China;National Center for TCM Inheritance and Innovation,Guangxi Key Laboratory of Medicinal Resources Protection and Genetic Improvement,National Engineering Research Center for Southwest Endangered Medicinal Materials Resources Development,Guangxi Botanical Garden of Medicinal Plants,Nanning 530023,China)

机构地区：[1]广西大学农学院,南宁530004 [2]广西药用植物园,国家中医药传承创新中心,广西药用资源保护与遗传改良重点实验室,西南濒危药材资源开发国家工程研究中心,南宁530023

出　　处：《植物病理学报》2024年第5期937-949,共13页Acta Phytopathologica Sinica

基　　金：中央引导地方科技发展专项(桂科ZY20111003);广西中医药管理局广西特色药材关键技术研究推广项目(GZKJ2314);国家自然科学基金项目(31860415);广西药用资源保护与遗传改良重点实验室资助项目(KL2022ZZ02)。

摘　　要：为了解广西罗汉果疑似病毒病病毒病原种类,为病毒病害的有效防治提供指导,利用小RNA测序分析了广西南宁广西大学实验田中罗汉果的病毒种类,并使用简并引物和特异性引物进行PCR验证;然后,使用病毒特异引物进行PCR或RT-PCR检测,调查广西罗汉果主产区病毒的分布情况;对罗汉果中新发现的病毒,克隆其基因组并分析其核苷酸序列特征。结果显示,小RNA测序和PCR验证均证实,广西大学罗汉果感染了菜豆金色花叶病毒属(Begomovirus)的中国南瓜曲叶病毒(squash leaf curl China virus,SLCCNV)和中国胜红蓟黄脉病毒(ageratum yellow vein China virus,AYVV)以及马铃薯Y病毒属(Potyvirus)的小西葫芦黄化花叶病毒(zucchini yellow mosaic virus,ZYMV)和番木瓜环斑病毒(papaya ringspot virus,PRSV);调查发现,桂林市永福县、龙胜县、临桂区和南宁市青秀区的罗汉果病样上均存在SLCCNV和ZYMV感染,桂林市临桂区、龙胜县的罗汉果上还检测到PRSV;克隆了SLCCNV-DNA-A基因组共2736 bp的全长序列和4个缺陷型分子、DNA-B共2648 bp的全长序列和2个缺陷型分子,以及AYVV共2745 bp的全长序列,三个全长序列分别编码7个、2个和7个蛋白质;序列分析表明,SLCCNV-DNA-A与中国云南地区的分离物关系最近,同源性最高,达99.56%,SLCCNV-DNA-B形成独立分支,与泰国的DNA-B序列同源性最高,达91.41%,SLCCNV-DNA-B的缺陷型分子含重组外源基因序列,AYVV与广西本地的苦苣菜分离物关系最近,同源性最高,达97.83%。表明罗汉果是begomovirus的新寄主,广西罗汉果病毒病害是begomovirus和potyvirus复合侵染引起。To clarify the types of viruses infecting Siraitia grosvenorii(Luo han guo)in Guangxi and provide instruction for the prevention,small RNA sequencing technology was employed to identify viruses,followed by PCR analysis to verify the results with degenerate and specific primer pairs.Then,the primers of corresponding viruses were used to investigate the distribution of virus species in the main production areas of S.grosvenorii in Guangxi.For these new viruses discovered in S.grosvenorii,all nucleotide sequences were characterized and phylogenetic trees were reconstructed,based on their amplified whole genomes.The results were as follows:according to small RNA sequencing and validation of polymerase chain reaction(PCR),leaves of S.grosvenoriifrom Guangxi University were infected with both squash leaf curl China virus(SLCCNV)and ageratum yellow vein China virus(AYVV)in Begomovirus,and both zucchini yellow mosaic virus(ZYMV)and papaya ringspot virus(PRSV)in Potyvirus.Further investigation revealed that all samples from Yongfu County,Lingui District and Longsheng County in Guilin and Qingxiu District in Nanning were infected with SLCCNV and ZYMV,and samples from Lingui District and Longsheng County in Guilin were also infected with PRSV.Their full-length genomes were all cloned,for SLCCNV-DNA-A(2736 bp)and its four defective molecules,DNA-B(2648 bp)and its two defective molecules,and AYVV(2745 bp),which encode seven,two,and seven proteins,respectively.Sequence alignment and phylogenetic analysis indicated that the sequence of SLCCNV-DNA-A exhibited the closest identity tothat of the isolate from Yunnan,China,with 99.56%identity;the sequence of SLCCNV-DNA-B exhibited the closest identity tothat of the isolate from Thailand,with 91.41%identity,whose defective molecules exhibited recombination with exogenous genes;and the sequence of AYVV exhibited the closest similarity to that of a common weed isolate in Guangxi,with 97.83%identity.These indicated that S.grosvenorii was a new host for viruses in Begomovirus,and S.grosv

关 键 词：罗汉果 菜豆金色花叶病毒属 马铃薯Y病毒属 基因组结构 

分 类 号：S432.1[农业科学—植物病理学]