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
金晓香 1姚俊汐 1尹跃艳 2李婷婷 3郭淼 2钟静 3和寿星 2陈越 3和家卫 2鄢波 1丁铭3
作者信息
- 1. 西南林业大学园林园艺学院,昆明 650224
- 2. 云南省农业科学院高山经济植物研究所,云南丽江 674100
- 3. 云南省农业科学院生物技术与种质资源研究所,昆明6502233
- 折叠
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,CymMV)、齿兰环斑病毒(odontoglossum ringspot virus,ORSV)和建兰褪绿花叶病毒(cymbidium chlorotic mosaic virus,CyCMV).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CyCMV复合侵染.为进一步明确ORSV和CyCMV云南分离物(ORSV-DH和CyCMV-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6 611 nt(Accession No.OP644547),CyCMV-DH 基因组全长序列为 4 083 nt(Accession NO.OP629812),将 ORSV-DH 和CyCMV-DH分别与己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％,CyCMV-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CyCMV-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)、CyCMV-Japan(NC_027123)亲缘关系最近.
Abstract
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 6 611 nt(GeneBank accession No.OP644547)and 4 083 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Key words
Cymbidium hybridum/ORSV/CyCMV/characterization of genomic sequence/mix-infection引用本文复制引用
基金项目
国家自然科学基金地区科学基金(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)
云南省科技重大专项(202302AE090005)
云南省农科院生物技术与种质资源研究所开放项目(2023005)
出版年
2024
NETL
NSTL
万方数据