侵染花叶青木的病毒种类鉴定及其基因序列分析  

Viruses Identification and Their Gene Sequences Analysis Infecting Aucuba japonica var.variegata

作　　者：梁海文 兰平秀 柳勤海 谭冠林 陈小姣 赵雁[2] 李凡[1] LIANG HaiWen;LAN PingXiu;LIU QinHai;TAN GuanLin;CHEN XiaoJiao;ZHAO Yan;LI Fan(College of Plant Protection,Yunnan Agricultural University,Kunming 650201;College of Horticulture and Landscape,Yunnan Agricultural University,Kunming 650201;Modern Education Technology Center,Yunnan Agricultural University,Kunming 650201)

机构地区：[1]云南农业大学植物保护学院,昆明650201 [2]云南农业大学园林园艺学院,昆明650201 [3]云南农业大学现代教育技术中心,昆明650201

出　　处：《中国农业科学》2023年第10期1893-1904,共12页Scientia Agricultura Sinica

基　　金：国家重点研发计划(2022YFD1400700);云南省专家工作站(202005AF150040)。

摘　　要：【目的】探明采自云南和海南表现为黄化、皱缩等表型的花叶青木病毒种类。【方法】首先,利用转录组测序技术对来自云南省昆明市的花叶青木样品进行检测;然后,根据转录组测序结果,采用RT-PCR对采自云南省昆明市和海南省海口市的73份花叶青木样品开展转录组测序结果中发现的病毒种类检测验证,并扩增获得相关病毒基因片段,对这些病毒的基因序列进行比对分析。【结果】转录组测序结果发现,送测的花叶青木样品中含有蚕豆萎蔫病毒2号(broad bean wilt virus 2,BBWV2)、菜豆黄花叶病毒(bean yellow mosaic virus,BYMV)、黄瓜花叶病毒(cucumber mosaic virus,CMV)、豌豆耳突花叶病毒1号(pea enation mosaic virus 1,PEMV-1)和豌豆耳突花叶病毒2号(pea enation mosaic virus 2,PEMV-2)5种病毒的侵染。但随后对来自云南和海南的73份花叶青木样品的RT-PCR验证中,只检测到BBWV2、BYMV、CMV和PEMV-24种病毒。其中,云南和海南的样品中均能检测到BBWV2,总检出率为11.0%;而BYMV、CMV和PEMV-2仅在云南的样品中被检测到,检出率分别为5.5%、1.4%和1.4%。从两地采集的花叶青木样品中既未发现PEMV-1的侵染,也未发现BBWV2、BYMV、CMV和PEMV-2的复合侵染。为了进一步分析BBWV2、BYMV、CMV和PEMV-2花叶青木分离物与来自其他寄主植物相关病毒分离物的分子变异,选取检测到上述4种病毒的样品,分别对BBWV2、BYMV和CMV 3种病毒的cp及PEMV-2的RdRp序列进行扩增,并对扩增获得的BBWV2490 nt的small cp(GenBank登录号:OQ137565、OQ137566)、BYMV 499 nt的cp核心区域(GenBank登录号:OQ137568)、CMV 880 nt的cp(GenBank登录号:OQ137567)和PEMV-2800nt的RdRp核心区域序列(GenBank登录号:OQ137569)进行分析。序列比对分析结果表明,BBWV2云南与海南花叶青木分离物间存在85.9%的核苷酸序列一致性,二者与GenBank中其他BBWV2分离物分别存在79.8%—94.5%和80.2%—92.0%的核苷酸序列一致性;BYMV、CMV【Objective】The objective of this study is to identify the viruses infecting Aucuba japonica var.variegata plants with symptoms of yellowing and shrinking which collected from Yunnan and Hainan provinces.【Method】Firstly,RNA sequencing was used to detect the viruses infecting A.japonica var.variegata plants from Kunming,Yunnan Province.Then,based on the RNA sequencing results,RT-PCR was used to detect these viruses in 73 A.japonica var.variegata field samples collected in Kunming of Yunnan Province and Haikou of Hainan Province,and the related virus gene fragments were amplified,sequenced and analyzed.【Result】RNA sequencing results showed that 5 viruses were detected from the pooled diseased plants,such as broad bean wilt virus 2(BBWV2),bean yellow mosaic virus(BYMV),cucumber mosaic virus(CMV),pea enation mosaic virus 1(PEMV-1),and pea enation mosaic virus 2(PEMV-2).However,only BBWV2,BYMV,CMV and PEMV-2 were detected in the 73 A.japonica var.variegata samples from Yunnan and Hainan.Among them,BBWV2 was detected in the A.japonica var.variegata samples both from Yunnan and Hainan,with total detection rate of 11.0%,while BYMV,CMV and PEMV-2 were only detected in the Yunnan samples with detection rate of 5.5%,1.4%and 1.4%,respectively.However,neither PEMV-1 nor the co-infections among BBWV2,BYMV,CMV and PEMV-2 was detected in these 73 A.japonica var.variegata samples from Yunnan or Hainan.In order to further analyze the molecular variation of BBWV2,BYMV,CMV and PEMV-2 aucuba isolates with the related virus isolates from other host plants,the 490 nt of small cp of BBWV2(accession number OQ137565,OQ137566),499 nt of the core cp region of BYMV(accession number OQ137568),880 nt of the cp of CMV(accession number OQ137567),and 800 nt of the core RdRp of PEMV-2(accession number OQ137569)were amplified from the A.japonica var.variegata samples infected with the above 4 viruses,and then the amplified gene sequences were analyzed,respectively.Sequence comparison analysis results showed that BBWV2 Yunnan aucuba isola

关 键 词：花叶青木 病毒鉴定 蚕豆萎蔫病毒2号 菜豆黄花叶病毒 黄瓜花叶病毒 豌豆耳突花叶病毒2号 

分 类 号：S432.41[农业科学—植物病理学]