蚕豆萎蔫病毒2号青海辣椒分离物的鉴定与全基因组序列克隆

doi:10.16420/j.issn.0513-353x.2021-0756

园艺学报 ›› 2023, Vol. 50 ›› Issue (1): 161-169.doi: 10.16420/j.issn.0513-353x.2021-0756

蚕豆萎蔫病毒2号青海辣椒分离物的鉴定与全基因组序列克隆

罗海林¹, 袁雷¹, 翁华¹, 闫佳会¹^,^*(), 郭青云¹^,^*(), 王文清², 马新明³

¹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，西宁 810016
²大通回族土族自治县蔬菜技术服务中心，西宁 810016
³循化撒拉族自治县农业农村和科技局，青海海东 810600

收稿日期:2022-05-12 修回日期:2022-08-13 出版日期:2023-01-25 发布日期:2023-01-18
通讯作者: *（E-mail：15209787170@163.com，guoqingyunqh@163.com）
基金资助:
青海省科技厅国际合作项目(2022-HZ-806);青海省农业有害生物综合治理重点实验室资助项目(2022-ZJ-Y10);青海省科技厅重点与成果转化项目(2019-NK-114)

Identification and Analysis of Complete Genomic Sequence of Broad Bean Wilt Virus 2 Pepper Isolate in Qinghai Province

LUO Hailin¹, YUAN Lei¹, WENG Hua¹, YAN Jiahui¹^,^*(), GUO Qingyun¹^,^*(), WANG Wenqing², MA Xinming³

¹Key Laboratory of Agricultural Integrated Pest Management，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
²Vegetable Technical Service Center of Datong County，Xining 810016，China
³Bureau of Agriculture and Rural & Science and Technology Affairs of Xunhua County，Haidong，Qinghai 810600，China

Received:2022-05-12 Revised:2022-08-13 Online:2023-01-25 Published:2023-01-18
Contact: *（E-mail：15209787170@163.com，guoqingyunqh@163.com）

摘要/Abstract

摘要：

对青海省循化县的辣椒病毒病进行调查，采集到部分叶片表现萎蔫和皱缩症状的样品，该症状与蚕豆萎蔫病毒2（BBWV2）引起的症状相似。为准确鉴定是否为与该病害相关的病毒，将采集的20个辣椒样品用特异性引物进行RT-PCR检测，其中13个样品检测结果为阳性。在20份循化县辣椒样品中，有13份样品被确认感染了蚕豆萎蔫病毒2（BBWV2）。基于核苷酸序列的系统发育分析表明，检测到的病毒与BBWV2聚类在一起，与其他地区或国家分离物的序列一致性较高，与蚕豆萎蔫病毒1（BBWV1）和野芝麻轻型花叶病毒（LMMV）分离物的序列一致性相对较低。结果表明该辣椒样品感染的病毒是BBWV2。

关键词: 辣椒, 蚕豆萎蔫病毒2, 全基因组

Abstract:

An investigation on pepper virus was conducted. Samples collected in Xunhua County，Qinghai Province shown symptoms of plant wilt and leaf shrinkage，which were very similar to the symptoms induced by broad bean wilt virus 2（BBWV2）. To identify the possible virus associated with the disease，20 samples were collected and detected with universal primer and specific primer using RT-PCR

and 13 samples shown positive results when using a pair of degenerate primers for BBWV2. The results showed that 13 of 20 pepper samples in Xunhua County were confirmed to be infected with broad bean wilting virus 2. Further phylogenetic analysis based on the nucleotide sequence also suggested that the virus detected was clustered with BBWV2 and shared the highest sequence identities with the isolates from abroad，and relatively lower identities with the isolates of broad bean wilt virus 1（BBWV1）or Lamium mild mosaic virus（LMMV）. All these results showed that the virus infected pepper is an isolate of BBWV2.

Key words: pepper, broad bean wilt virus, whole genome

中图分类号:

S641.3

罗海林, 袁雷, 翁华, 闫佳会, 郭青云, 王文清, 马新明. 蚕豆萎蔫病毒2号青海辣椒分离物的鉴定与全基因组序列克隆[J]. 园艺学报, 2023, 50(1): 161-169.

LUO Hailin, YUAN Lei, WENG Hua, YAN Jiahui, GUO Qingyun, WANG Wenqing, MA Xinming. Identification and Analysis of Complete Genomic Sequence of Broad Bean Wilt Virus 2 Pepper Isolate in Qinghai Province[J]. Acta Horticulturae Sinica, 2023, 50(1): 161-169.

图/表 6

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病毒序列 Virus sequence	引物名称 Name	引物序列（5′-3′） Sequence	退火温度/℃ T_m	片段大小/bp Length	文献 Reference
BBWV2	BBWVdF	AGRTATATGCTTGGGCAAGCGCATG	60	490	刘勇等， 2019
	BBWVdR	CATGAACATTCCCCATCTCCACGTG			刘勇等， 2019
RNA1	BBWV2 1-F	AAACAAACAGCTTTCGTTCCG	54	1 080	Kwak et 2013b
	BBWV2 1-R	CTGAAATAGCTGCCATAGCAG
	BBWV2 1-F2	TTTGCAGCGTGGATGCC	55	1 075
	BBWV2 1-R2	CTTGTAGCCTGACCAAAAAGG
	BBWV2 1-F3	SGCAGATTGGTTATSWGAR	53	880
	BBWV2 1-R3	ACCGTGAGRCCCACWAR
	BBWV2 1-F4	MGRCGAAGRCACGCAGTRRT	57	1 083
	BBWV2 1-R4	AAGCACACTTGTGTYCTMACAT
	BBWV2 1-F5	CAAAGTGATRTKCCTAAYC	48	1 561
	BBWV2 1-R5	CCRTCAAAYCTGCTGTAA
	BBWV2 1-F6	CGSCTWGATCGTGGTCC	54	1 766
	BBWV2 1-R6	TTCCCTCACTACTGAAATTTACTC
RNA2	BBWV2 2-1F	CAAACAGCTTTCGTTCCGAA	52	927	Kwak et 2013b
	BBWV2 2-1R	AAGCCATCTGCCACCAATTC
	BBWY2 2-1d	CGCATA(G)GTC(T)TCTT(C)GTA(G)GCA(T)GA	55	900
	BBWV2 2-2F	GCCATGTCAGGTGCTGGA
	BBWV2 2-2U	TACAAT(C)TTG(A)CCTGGG(A)CCTAGC	56	1 119
	BBMV2 2-2r	CATGTTCCCAACCTTTTCAATGT
	BBWV2 2-3U	ATTT(C)TA(G)GATTTTACT(C)CTC(T)ATGAG	55	1 163
	BBWV2 2-3d	AGGTCATGGAACCCATTTTAATGG

病毒序列 Virus sequence	引物名称 Name	引物序列（5′-3′） Sequence	退火温度/℃ T_m	片段大小/bp Length	文献 Reference
BBWV2	BBWVdF	AGRTATATGCTTGGGCAAGCGCATG	60	490	刘勇等， 2019
	BBWVdR	CATGAACATTCCCCATCTCCACGTG			刘勇等， 2019
RNA1	BBWV2 1-F	AAACAAACAGCTTTCGTTCCG	54	1 080	Kwak et 2013b
	BBWV2 1-R	CTGAAATAGCTGCCATAGCAG
	BBWV2 1-F2	TTTGCAGCGTGGATGCC	55	1 075
	BBWV2 1-R2	CTTGTAGCCTGACCAAAAAGG
	BBWV2 1-F3	SGCAGATTGGTTATSWGAR	53	880
	BBWV2 1-R3	ACCGTGAGRCCCACWAR
	BBWV2 1-F4	MGRCGAAGRCACGCAGTRRT	57	1 083
	BBWV2 1-R4	AAGCACACTTGTGTYCTMACAT
	BBWV2 1-F5	CAAAGTGATRTKCCTAAYC	48	1 561
	BBWV2 1-R5	CCRTCAAAYCTGCTGTAA
	BBWV2 1-F6	CGSCTWGATCGTGGTCC	54	1 766
	BBWV2 1-R6	TTCCCTCACTACTGAAATTTACTC
RNA2	BBWV2 2-1F	CAAACAGCTTTCGTTCCGAA	52	927	Kwak et 2013b
	BBWV2 2-1R	AAGCCATCTGCCACCAATTC
	BBWY2 2-1d	CGCATA(G)GTC(T)TCTT(C)GTA(G)GCA(T)GA	55	900
	BBWV2 2-2F	GCCATGTCAGGTGCTGGA
	BBWV2 2-2U	TACAAT(C)TTG(A)CCTGGG(A)CCTAGC	56	1 119
	BBMV2 2-2r	CATGTTCCCAACCTTTTCAATGT
	BBWV2 2-3U	ATTT(C)TA(G)GATTTTACT(C)CTC(T)ATGAG	55	1 163
	BBWV2 2-3d	AGGTCATGGAACCCATTTTAATGG

国家 Country	寄主 Host	分离物 Isolate	RNA1		RNA2
国家 Country	寄主 Host	分离物 Isolate	核苷酸/% Nucleotide	氨基酸/% Amion acid	核苷酸/% Nucleotide	氨基酸/% Amion acid
韩国Korea	蚕豆 Vicia faba	BBWV2-BB9	100	89.92	100	84.34
	菠菜 Spinacia oleracea	BBWV2-SP2	100	90.02	100	84.13
		BBWV2-SP1	100	90.00	100	84.19
		BBWV2-SP	100	89.97	100	84.72
	牛膝 Achyranthes bidentata Blume	BBWV2-AB1	100	90.24	100	86.97
	辣椒 Capsicum annuum	BBWV2-RP2	100	90.24	100	84.04
	辣椒	BBWV2-PAP1	100	89.95	100	84.80
	C. annuum var. gulosum	BBWV2-PAP3	100	91.48	100	84.91
	薯蓣 Dioscorea oppositifolia	BBWV2-BR1	99	77.44	99	87.16
中国China	蚕豆 Vicia faba	BBWV2-B935	99	77.58	100	84.51
	薯蓣 Dioscorea opposita	BBWV2-SG1	99	77.32	100	87.09
	辣椒 Capsicum annuum L.	BBWV2-Ca	99	77.36	99	82.41
	番茄 Lycopersicon esculentum	BBWV2-XJ14-3	99	77.50	100	87.18
日本Japan	三花龙胆 Gentiana triflora	BBWV2-Ty	99	77.08	99	88.61
	蚕豆 Vicia faba	BBWV2-MB7	99	78.03	100	85.90
西班牙Spain	辣椒 Capsicum annuum	BBWV1-Ben	74	65.68	62	64.99
	蚕豆 Vicia faba	BBWV1-PBenR1	74	66.01	62	65.16
英国 United Kingdom	块根落葵 Ullucus tuberosus	BBWV1-UL1	73	65.88	52	66.61

国家 Country	寄主 Host	分离物 Isolate	RNA1		RNA2
国家 Country	寄主 Host	分离物 Isolate	核苷酸/% Nucleotide	氨基酸/% Amion acid	核苷酸/% Nucleotide	氨基酸/% Amion acid
韩国Korea	蚕豆 Vicia faba	BBWV2-BB9	100	89.92	100	84.34
	菠菜 Spinacia oleracea	BBWV2-SP2	100	90.02	100	84.13
		BBWV2-SP1	100	90.00	100	84.19
		BBWV2-SP	100	89.97	100	84.72
	牛膝 Achyranthes bidentata Blume	BBWV2-AB1	100	90.24	100	86.97
	辣椒 Capsicum annuum	BBWV2-RP2	100	90.24	100	84.04
	辣椒	BBWV2-PAP1	100	89.95	100	84.80
	C. annuum var. gulosum	BBWV2-PAP3	100	91.48	100	84.91
	薯蓣 Dioscorea oppositifolia	BBWV2-BR1	99	77.44	99	87.16
中国China	蚕豆 Vicia faba	BBWV2-B935	99	77.58	100	84.51
	薯蓣 Dioscorea opposita	BBWV2-SG1	99	77.32	100	87.09
	辣椒 Capsicum annuum L.	BBWV2-Ca	99	77.36	99	82.41
	番茄 Lycopersicon esculentum	BBWV2-XJ14-3	99	77.50	100	87.18
日本Japan	三花龙胆 Gentiana triflora	BBWV2-Ty	99	77.08	99	88.61
	蚕豆 Vicia faba	BBWV2-MB7	99	78.03	100	85.90
西班牙Spain	辣椒 Capsicum annuum	BBWV1-Ben	74	65.68	62	64.99
	蚕豆 Vicia faba	BBWV1-PBenR1	74	66.01	62	65.16
英国 United Kingdom	块根落葵 Ullucus tuberosus	BBWV1-UL1	73	65.88	52	66.61

蚕豆萎蔫病毒2号青海辣椒分离物的鉴定与全基因组序列克隆

Identification and Analysis of Complete Genomic Sequence of Broad Bean Wilt Virus 2 Pepper Isolate in Qinghai Province

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蚕豆萎蔫病毒2号青海辣椒分离物的鉴定与全基因组序列克隆

Identification and Analysis of Complete Genomic Sequence of Broad Bean Wilt Virus 2 Pepper Isolate in Qinghai Province

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可视化

摘要/Abstract

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图/表 6

参考文献 27

相关文章 15

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