侵染青海辣椒的辣椒隐症病毒2基因组克隆与分析

doi:10.3969/j.issn.1004-1524.2023.06.14

浙江农业学报 ›› 2023, Vol. 35 ›› Issue (6): 1368-1374.DOI: 10.3969/j.issn.1004-1524.2023.06.14

侵染青海辣椒的辣椒隐症病毒2基因组克隆与分析

罗海林(), 袁雷, 闫佳会^*(), 郭青云

青海大学农林科学院,青海省农业有害生物综合治理重点实验室,农业农村部西宁作物有害生物科学观测实验站,青海西宁 810016

收稿日期:2022-05-12 出版日期:2023-06-25 发布日期:2023-07-04
通讯作者: *闫佳会,E-mail:15209787170@163.com
作者简介:罗海林(1997—),女,安徽六安人,硕士研究生,主要从事辣椒病毒病研究。E-mail:1369507819@qq.com
基金资助:
青海省农业有害生物综合治理重点实验室(2022-ZJ-Y10);青海省国际科技合作项目(2022-HZ-806)

Cloning and identification of Pepper cryptic virus 2 in Qinghai Province, China

LUO Hailin(), YUAN Lei, YAN Jiahui^*(), GUO Qingyun

Key Laboratory of Agricultural Integrated Pest Management of Qinghai Province, Scientific Observing and Experimental Station of Crop Pest in Xining, Ministry of Agriculture and Rural Affairs, Academy of Agriculture and Forestry Sciences, Qinghai University, Xining 810016, China

Received:2022-05-12 Online:2023-06-25 Published:2023-07-04

摘要/Abstract

摘要：

辣椒隐症病毒2(Pepper cryptic virus 2,PCV2)是近年来发现的辣椒新病毒之一,常与其他病毒复合侵染,影响辣椒的产量和品质。2021年从青海海东市采集疑似病样叶片,采用RT-PCR和基因克隆等方法,获得PCV2青海分离物的两条dsRNA,序列全长分别为1 579、1 435 bp。核苷酸相似性比对发现,PCV2青海分离物两条RNA序列与已报道的其他地区PCV2分离物的序列相似性高于95%,系统进化分析结果显示,PCV2青海分离物的两条RNA均与PCV2分离物Chiltepin 53处于独立的分支,结果表明,青海辣椒上检测得到的病毒为辣椒隐症病毒2。将其与PCV2-QH和PCV2-XJ的核苷酸序列进行分析,结果显示,二者存在多位点差异。本研究首次报道PCV2在青海发生。

关键词: 辣椒, 辣椒隐症病毒2(PCV2), 全基因组, 序列分析

Abstract:

Pepper cryptic virus 2 is one of the new viruses in pepper. It often infects with other viruses, seriously reducing the yield and quality of pepper. In 2021, suspected pepper’s leaf samples were collected from Haidong, Qinghai. Two dsRNA complete gene sequences of PCV2 were obtained by using RT-PCR and gene cloning. The two-length sequence of Qinghai isolates were 1 579 bp and 1 435 bp. The nucleotide sequence identities against other Pepper cryptic virus 2 isolates were over 95%. Phylogenetic analysis showed that PCV2 Qinghai isolate clustered with Chiltepin53 in a sub-clade. All these results showed that the virus infected pepper was an isolate of PCV2. Analyzing the nucleotide sequence of PCV2-QH and PCV2-XJ, the results showed that there were multiple differences. This is the first report of PCV2 infecting pepper in Qinghai.

Key words: pepper, Pepper cryptic virus 2, complete gene sequence, sequence analysis

中图分类号:

S436.418

罗海林, 袁雷, 闫佳会, 郭青云. 侵染青海辣椒的辣椒隐症病毒2基因组克隆与分析[J]. 浙江农业学报, 2023, 35(6): 1368-1374.

LUO Hailin, YUAN Lei, YAN Jiahui, GUO Qingyun. Cloning and identification of Pepper cryptic virus 2 in Qinghai Province, China[J]. Acta Agriculturae Zhejiangensis, 2023, 35(6): 1368-1374.

图/表 7

参考文献 22

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基因/片段 Gene/Segment	退火温度 Tm/℃	目的片段长度 Length/bp	引物名称 Primer name	引物序列 Primer sequence(5'→3')
PVC2	59.0	375	PCV2F	TCATCCGTCCAGCTAACGTA
			PCV2R	CGTCTCTTTTCTGAGCGGTA
RNA1	57.8	1 579	PCV2_RNA1-F	CAGTATCGAGAGATCACTGTGAC
			PCV2_RNA1-R	GGTTAGTGCCTTATGCCCA
RNA2	58.3	1 435	PCV2_RNA2-F	AGAATTTTCTGATCACAGTATCAAGAGATCACTG
			PCV2_RNA2-R	GACAAGGGGTACTAACCTCTTG

基因/片段 Gene/Segment	退火温度 Tm/℃	目的片段长度 Length/bp	引物名称 Primer name	引物序列 Primer sequence(5'→3')
PVC2	59.0	375	PCV2F	TCATCCGTCCAGCTAACGTA
			PCV2R	CGTCTCTTTTCTGAGCGGTA
RNA1	57.8	1 579	PCV2_RNA1-F	CAGTATCGAGAGATCACTGTGAC
			PCV2_RNA1-R	GGTTAGTGCCTTATGCCCA
RNA2	58.3	1 435	PCV2_RNA2-F	AGAATTTTCTGATCACAGTATCAAGAGATCACTG
			PCV2_RNA2-R	GACAAGGGGTACTAACCTCTTG

国家 Country	寄主 Host	分离物 Isolate	RNA1登录号 Accession No.	核苷酸序列一致性 Nucleotide Similarities/%	氨基酸序列一致性 Amion acids Similarities/%	RNA 2登录号 Accession No.	核苷酸序列一致性 Nucleotide Similarities/%	氨基酸序列一致性 Amion acids Similarities/%
中国China	Capsicum annuum	PCV2-XJ	MT328005.1	99	99.21	MT328056.1	99	99.64
多米尼加共和国	Capsicum annuum	DR	KX525268.1	99	99.02	KX525269.1	99	99.50
Dominican Republic
韩国Korea	Capsicum annuum	SC10-2	KY923703.1	99	98.89	KY923704.1	99	99.50
韩国Korea	Ixais dentate NAKAI	YY	LC195294.1	99	98.36	LC195295.1	99	99.21
土耳其Turkey	Capsicum annuum	Tur	LC325817.1	99	96.00	LC325818	99	97.99
斯洛伐克	Capsicum annuum	N65	MN723540.1	99	95.96	MN723541.1	99	97.92
Slovakia
美国USA	Capsicum annuum	HW-01	JN117278.1	99	95.94	JN117279.1	99	97.92
中国China	Capsicum annuum	PCV2-CQ	KX905077.1	78	98.92	KX905078.1	99	99.50
美国USA	Capsicum annuum	Ember	MG542626.1	65	99.11	MG542618.1	83	99.83

国家 Country	寄主 Host	分离物 Isolate	RNA1登录号 Accession No.	核苷酸序列一致性 Nucleotide Similarities/%	氨基酸序列一致性 Amion acids Similarities/%	RNA 2登录号 Accession No.	核苷酸序列一致性 Nucleotide Similarities/%	氨基酸序列一致性 Amion acids Similarities/%
中国China	Capsicum annuum	PCV2-XJ	MT328005.1	99	99.21	MT328056.1	99	99.64
多米尼加共和国	Capsicum annuum	DR	KX525268.1	99	99.02	KX525269.1	99	99.50
Dominican Republic
韩国Korea	Capsicum annuum	SC10-2	KY923703.1	99	98.89	KY923704.1	99	99.50
韩国Korea	Ixais dentate NAKAI	YY	LC195294.1	99	98.36	LC195295.1	99	99.21
土耳其Turkey	Capsicum annuum	Tur	LC325817.1	99	96.00	LC325818	99	97.99
斯洛伐克	Capsicum annuum	N65	MN723540.1	99	95.96	MN723541.1	99	97.92
Slovakia
美国USA	Capsicum annuum	HW-01	JN117278.1	99	95.94	JN117279.1	99	97.92
中国China	Capsicum annuum	PCV2-CQ	KX905077.1	78	98.92	KX905078.1	99	99.50
美国USA	Capsicum annuum	Ember	MG542626.1	65	99.11	MG542618.1	83	99.83

片段 Segments	序列位点 Sequence position	PCV2-QH	PCV2-XJ	蛋白/非编码区 Protein/Untranslated region	突变 Mutation
dsRNA1	247	C	T	RNA依赖的RNA聚合酶 RNA dependent RNA polymerase(RdRp)	非同义替换 Nonsynonymous substitution
	478	C	T	RNA依赖的RNA聚合酶 RNA dependent RNA polymerase(RdRp)	非同义替换 Nonsynonymous substitution
	679	A	G	RNA依赖的RNA聚合酶 RNA dependent RNA polymerase(RdRp)	非同义替换 Nonsynonymous substitution
	1 039	G	T	RNA依赖的RNA聚合酶 RNA dependent RNA polymerase(RdRp)	非同义替换 Nonsynonymous substitution
	1 055	G	T	RNA依赖的RNA聚合酶 RNA dependent RNA polymerase(RdRp)	非同义替换 Nonsynonymous substitution
	1 060	G	T	RNA依赖的RNA聚合酶 RNA dependent RNA polymerase(RdRp)	非同义替换 Nonsynonymous substitution
	1 083	G	T	RNA依赖的RNA聚合酶 RNA dependent RNA polymerase(RdRp)	同义替换 Synonymous substitution
dsRNA2	51	G	A	非编码区Non-coding region	—
	52	A	G	非编码区Non-coding region	—
	53	T	A	非编码区Non-coding region	—
	54	A	C	非编码区Non-coding region	—
	55	C	G	非编码区Non-coding region	—
	450	C	T	外壳蛋白Coat protein(CP)	同义替换Synonymous substitution
	1 431	A	T	非编码区Non-coding region	—
	1 433	T	A	非编码区Non-coding region	—
	1 435	C	T	非编码区Non-coding region	—
	1 436	G	T	非编码区Non-coding region	—
	1 437	C	T	非编码区Non-coding region	—

侵染青海辣椒的辣椒隐症病毒2基因组克隆与分析

Cloning and identification of Pepper cryptic virus 2 in Qinghai Province, China

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