云南大花蕙兰复合感染的ORSV和CyCMV病毒基因组序列特征分析  

Molecular Characterization of the Odontoglossum Ringspot Virus and Cymbidium Chlorotic Mosaic Virus Mix-Infecting Cymbidium hybridum in Yunnan

作　　者：金晓香 姚俊汐 尹跃艳[3] 李婷婷[2] 郭淼[3] 钟静[2] 和寿星[3] 陈越[2] 和家卫 鄢波[1] 丁铭[2] JIN Xiaoxiang;YAO Junxi;YIN Yueyan;LI Tingting;GUO Miao;ZHONG Jing;HE Shouxing;CHEN Yue;HE Jiawei;YAN Bo;DING Ming(Department of Landscape and Horticulture,Southwest Forestry University,Kunming 650224,China;Institute of Biotechnology and Germplasm Resources,Yunnan Academy of Agricultural Sciences,Kunming 650223,China;Institute of Alpine Economic Plants,Yunnan Academy of Agricultural Sciences,Lijiang,Yunnan 674100,China)

机构地区：[1]西南林业大学园林园艺学院,昆明650224 [2]云南省农业科学院生物技术与种质资源研究所,昆明6502233 [3]云南省农业科学院高山经济植物研究所,云南丽江674100

出　　处：《园艺学报》2024年第8期1844-1852,共9页Acta Horticulturae Sinica

基　　金：国家自然科学基金地区基金项目(32360644);云南省种子种业联合实验室项目(202205AR070001-02);云南省乡村振兴科技专项(200202A);云南省重大科技专项计划项目(202302AE090005);云南省农业科学院生物技术与种质资源研究所开放项目(2023005)。

摘　　要：从云南丽江采集了40个大花蕙兰(Cymbidium hybridum)感病样品,利用高通量转录组测序对其中的病毒种类进行分析,Blastn N比对发现样品中存在建兰花叶病毒(cymbidium mosaic virus,Cym MV)、齿兰环斑病毒(odontoglossum ringspot virus,ORSV)和建兰褪绿花叶病毒(cymbidium chlorotic mosaic virus,Cy CMV)。利用RT-PCR对样品中病毒分离物的CP序列进行扩增,10份样品为ORSV和Cy CMV复合侵染。为进一步明确ORSV和Cy CMV云南分离物(ORSV-DH和Cy CMV-DH)的序列特征,分别对其全基因组序列进行了扩增及克隆,结果表明:ORSV-DH基因组全长序列为6611 nt(Accession No.OP644547),Cy CMV-DH基因组全长序列为4083 nt(Accession NO.OP629812),将ORSV-DH和Cy CMV-DH分别与已报道的分离物的全基因组序列进行核苷酸序列分析,发现ORSV-DH与其他分离物的核苷酸相似性为97.69%~99.41%,Cy CMV-DH为89.84%~98.19%。根据病毒全基因组核苷酸序列分别构建了ORSV-DH和Cy CMV-DH与本属其他分离物的系统进化树,明确了其分别与日本分离物ORSV-Cy-1(S83257)、Cy CMV-Japan(NC_027123)亲缘关系最近。Forty greenhouse-grown orchid samples were collected in Lijiang City,Yunnan Province,China,to assess the presence of viruses.Total RNAs from two samples were extracted and subjected them to high-throughput transcriptome sequencing(HTS)using the Illumina HiSeq platform.BLASTN searches revealed the presence of contigs with high similarities to cymbidium mosaic virus(CymMV),odontoglossum ringspot virus(ORSV),and cymbidium chlorotic mosaic virus(CyCMV).RT-PCR assays were conducted to confirm the presence of these viruses in the individual collected samples,and 10 samples were found to be mix-infected by ORSV and CyCMV.To further ascertain their molecular characteristics,the complete genomic sequences of the ORSV-DH and CyCMV-DH isolates were cloned.The full-length sequences of the ORSV-DH and CyCMV-DH genomes were determined to be 6611 nt(GeneBank accession No.OP644547)and 4083 nt(GeneBank accession No.OP629812),respectively.Sequence alignments revealed that ORSV-DH displayed nucleotide sequence identities of 97.69%–99.41%with previously reported ORSV isolates,whereas CyCMV-DH showed identities of 89.84%–98.19%with other CyCMV isolates.Phylogenetic trees of ORSV-DH and CyCMV-DH based on their complete genomic sequences were constructed and showed closely related to the Japanese isolate ORSV-Cy-1(S83257)and the Japanese isolate CyCMV-Japan(NC_027123),respectively.

关 键 词：大花蕙兰 齿兰环斑病毒 建兰褪绿花叶病毒 基因组序列特征 复合侵染 

分 类 号：S682.31[农业科学—观赏园艺]