青海省循化县辣椒脉斑驳病毒分离物的全基因组序列克隆与序列分析

doi:10.3969/j.issn.1004-1524.20240564

浙江农业学报 ›› 2025, Vol. 37 ›› Issue (3): 612-620.DOI: 10.3969/j.issn.1004-1524.20240564

青海省循化县辣椒脉斑驳病毒分离物的全基因组序列克隆与序列分析

温枭雄¹^,²(), 舒琴¹^,², 鲍梦楠¹^,², 闫佳会¹^,²^,^*()

1.青海大学农林科学院植物保护研究所,青海西宁 810016
2.青海省农业有害生物综合治理重点实验室,青海西宁 810016

收稿日期:2024-06-28 出版日期:2025-03-25 发布日期:2025-04-02
作者简介:温枭雄(1999—),男,山东青岛人,硕士研究生,研究方向为植物病毒。E-mail: 15863005348@163.com
通讯作者: * 闫佳会,E-mail:15209787170@163.com
基金资助:
青海省中央引导地方科技发展基金(2024ZY025)

Identification and complete genome sequence cloning of the chili veinal mottle virus in Qinghai Province of China

WEN Xiaoxiong¹^,²(), SHU Qin¹^,², BAO Mengnan¹^,², YAN Jiahui¹^,²^,^*()

1. Institute of Plant Protection, Academy of Agricultural and Forestry Sciences, Qinghai University,Xining 810016, China
2. Key Laboratory of Integrated Pest Management in Agriculture of Qinghai Province, Xining 810016, China

Received:2024-06-28 Online:2025-03-25 Published:2025-04-02

摘要/Abstract

摘要：

为调查青海省海东市循化县辣椒病毒病的发生情况,探究引起辣椒皱缩、矮小等症状的原因,本研究于2023年9月采集了循化县辣椒种植区域的疑似病毒病样本,利用reverse transcription-polymerase chain reaction(RT-PCR)技术检测。结果显示,19份辣椒病样中,8份样本被确认为感染辣椒脉斑驳病毒(Chilli Veinal Mottle virus, ChiVMV),检出率为42%。该病毒的RNA链全长为9 754 nt。全基因组序列克隆和分析揭示,青海分离物的序列与其他ChiVMV分离物相比存在一个显著的重组事件,这一重组事件导致了部分基因组区域的序列差异,与中国的合肥、宁波、昆明、泸州、苏州,以及印度和巴基斯坦等地的分离物相比,本研究获得的青海分离物的核苷酸序列一致性介于84%~100%。本研究首次在青海省循化县辣椒上检测到ChiVMV,并提供了该病毒的全基因组序列,这为ChiVMV抗病品种选育和相关病毒病防治提供了新的理论依据和数据支持,并有助于理解病毒的遗传变异及其在不同地域的流行动态。

关键词: 全基因组, 辣椒脉斑驳病毒, RT-PCR

Abstract:

To assess the occurrence of viral diseases in chili peppers in Xunhua County, Haidong City, Qinghai Province, and to investigate the factors contributing to symptoms such as leaf curling and stunting, this study collected suspected viral samples from chili cultivation areas in September 2023. Utilizing reverse transcription-polymerase chain reaction (RT-PCR) for detection, results indicated that 8 out of 19 symptomatic chili samples were confirmed to be infected with Chilli veinal mottle virus(ChiVMV), yielding a detection rate of 42%. The complete RNA genome of the virus was determined to be 9 754 nt in length. Genomic sequencing and analysis revealed a significant recombination event in the Qinghai isolate, leading to sequence variations in certain genomic regions when compared to other ChiVMV isolates. The nucleotide sequence identity of the Qinghai isolate ranged from 84% to 100% compared to isolates from Hefei, Ningbo, Kunming, Luzhou, and Suzhou in China, as well as those from India and Pakistan. This study represents the first detection of ChiVMV in chili peppers in Xunhua County, Qinghai Province, and provides the complete genomic sequence of the virus. These findings contribute valuable insights and empirical data for the development of resistant cultivars and the management of related viral diseases, thereby enhancing our understanding of viral genetic variation and its epidemiological dynamics across different regions.

Key words: complete genome, ChiVMV, RT-PCR

中图分类号:

温枭雄, 舒琴, 鲍梦楠, 闫佳会. 青海省循化县辣椒脉斑驳病毒分离物的全基因组序列克隆与序列分析[J]. 浙江农业学报, 2025, 37(3): 612-620.

WEN Xiaoxiong, SHU Qin, BAO Mengnan, YAN Jiahui. Identification and complete genome sequence cloning of the chili veinal mottle virus in Qinghai Province of China[J]. Acta Agriculturae Zhejiangensis, 2025, 37(3): 612-620.

图/表 8

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引物名称 Primer name	引物序列 Primer sequences (5'→3')	退火温度 Tm/℃	长度 Length/bp
ChiVMV-F	TGACTATGATGTCACAGATGA	57.5	795
ChiVMV-R	AACGCCAACTATTGAACAGCTT	58.0
1F	TCATAAAATTTTCTTGAAATTCAGTCATCAG	55.0	1 451
1R	CTTGTTCCTAAAAAGATGTAAGGAACCT	57.0
2F	ATCATGAAGCAACGGGTAAAGT	57.5	2 500
2R	CCTTGACATATTGAATGAACTCTACCTC	58.0
3F	GGAGAAGATAGCATTAGTTCCAGTACCACT	61.0	2 500
3R	TAGTACATTGATTGTATTCACGCACTGACAT	60.0
4F	GAACCATGGCACTTTTTGAGCTTCC	61.0	2 500
4R	GCCATTCCAAAGTCTCAACCTTTGC	61.5
5F	CTTTCTCCGAACTTGGTTTGTCGTATGATTTC	61.0	1 553
5R	ATAACATATATAAAGTAGTATGAAACCGAGCGTGC	60.0

引物名称 Primer name	引物序列 Primer sequences (5'→3')	退火温度 Tm/℃	长度 Length/bp
ChiVMV-F	TGACTATGATGTCACAGATGA	57.5	795
ChiVMV-R	AACGCCAACTATTGAACAGCTT	58.0
1F	TCATAAAATTTTCTTGAAATTCAGTCATCAG	55.0	1 451
1R	CTTGTTCCTAAAAAGATGTAAGGAACCT	57.0
2F	ATCATGAAGCAACGGGTAAAGT	57.5	2 500
2R	CCTTGACATATTGAATGAACTCTACCTC	58.0
3F	GGAGAAGATAGCATTAGTTCCAGTACCACT	61.0	2 500
3R	TAGTACATTGATTGTATTCACGCACTGACAT	60.0
4F	GAACCATGGCACTTTTTGAGCTTCC	61.0	2 500
4R	GCCATTCCAAAGTCTCAACCTTTGC	61.5
5F	CTTTCTCCGAACTTGGTTTGTCGTATGATTTC	61.0	1 553
5R	ATAACATATATAAAGTAGTATGAAACCGAGCGTGC	60.0

登录号 Accession no.	分离物 Isolate	寄主 Host	地区 Country	核苷酸相似度 Nucleotide Similiarity/%
KX650863	Dzh-Qyg	Nicotiana tabacum	China: Hefei	99.97
ON168942	ChiVMV-D	Capsicum annuum	China: Ningbo	93.34
MK405594	SiChuan Luzhou	Nicotiana tabacum	China: Luzhou	93.19
OP589298	GZ	Tobacco	China: Guiyang	93.11
MT787292	ChiVMV-Dh	Chilli pepper	China: Kunming	92.23
PP582383	AH	Capsicum	China: Suzhou	87.36
OP404087	YunN	Capsicum annuum	China: Ningbo	85.75
ON838424	-	Chilli	China: Ningbo	85.64
MN207122	ChiVMV-PK	Capsicum annuum	Pakistan	86.12
MN508960	ChiVMV-Uravakonda	Hot pepper	India: Uravakonda, Andhra pradesh	84.41
OR347840	To	Tomato	India: Karnataka	84.29

登录号 Accession no.	分离物 Isolate	寄主 Host	地区 Country	核苷酸相似度 Nucleotide Similiarity/%
KX650863	Dzh-Qyg	Nicotiana tabacum	China: Hefei	99.97
ON168942	ChiVMV-D	Capsicum annuum	China: Ningbo	93.34
MK405594	SiChuan Luzhou	Nicotiana tabacum	China: Luzhou	93.19
OP589298	GZ	Tobacco	China: Guiyang	93.11
MT787292	ChiVMV-Dh	Chilli pepper	China: Kunming	92.23
PP582383	AH	Capsicum	China: Suzhou	87.36
OP404087	YunN	Capsicum annuum	China: Ningbo	85.75
ON838424	-	Chilli	China: Ningbo	85.64
MN207122	ChiVMV-PK	Capsicum annuum	Pakistan	86.12
MN508960	ChiVMV-Uravakonda	Hot pepper	India: Uravakonda, Andhra pradesh	84.41
OR347840	To	Tomato	India: Karnataka	84.29

青海省循化县辣椒脉斑驳病毒分离物的全基因组序列克隆与序列分析

Identification and complete genome sequence cloning of the chili veinal mottle virus in Qinghai Province of China

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