番茄早疫病菌dsRNA多样性分析

doi:10.6048/j.issn.1001-4330.2023.12.022

新疆农业科学 ›› 2023, Vol. 60 ›› Issue (12): 3057-3064.DOI: 10.6048/j.issn.1001-4330.2023.12.022

• 畜牧兽医·微生物·农业装备工程与机械化 • 上一篇下一篇

番茄早疫病菌dsRNA多样性分析

刘升学¹(), 李思琪², 王晓东², 杨德松²()

1.石河子大学分析测试中心,新疆石河子 832000
2.石河子大学农学院/新疆绿洲农业病虫害治理与植保资源利用重点实验室,新疆石河子 832003

收稿日期:2023-03-21 出版日期:2023-12-20 发布日期:2024-01-03
通信作者: 杨德松(1977-),男,安徽滁州人,教授,博士,研究方向为农药环境毒理学, (E-mail)yds_agr@shzu.edu.cn
作者简介:刘升学(1974-),男,甘肃通渭人,副教授,博士,研究方向植物病理学,(E-mail)liushengxue-1@163.com
基金资助:
新疆生产建设兵团应用基础研究项目“加工番茄早疫病真菌病毒的鉴定及其生防菌病毒的筛选”(2015AG010)

Diversity analysis of dsRNA in Alternaria solani

LIU Shengxue¹(), LI Siqi², WANG Xiaodong², YANG Desong²()

1. Analysis and Testing Center, Shihezi University, Shihezi Xinjiang 832000,China
2. College of Agriculture, Shihezi University,Key Laboratory for Oasis Agricultural Pest Management and Plant Resource Utilization, Shihezi Xinjiang 832000,China

Received:2023-03-21 Online:2023-12-20 Published:2024-01-03
Correspondence author: YANG Desong(1977-),male, Doctor, Professor, Research direction: Pesticide toxicology, (E-mail)yds_agr@shzu.edu.cn
Supported by:
Basic R & D Project of XPCC "Identification of Processed Tomato Early Blight Fungal Viruses and Screening of Biocontrol Viruses"(2015AG010)

摘要/Abstract

摘要：

【目的】研究番茄早疫病菌携带dsRNA的多样性,为筛选具有生防作用的真菌病毒提供依据。【方法】从石河子周边加工番茄种植区采集典型症状的组织样品,通过组织分离和单孢分离法获得纯培养;提取 dsRNA及琼脂糖凝胶电泳明确携带dsRNA 的情况。选择6个菌株,通过随机引物RT-PCR、序列分析,研究Blast对携带病毒种类及分类。【结果】从分离到的191株番茄早疫病菌菌株中提取dsRNA,有43株中检测到1-4条dsRNA条带,带毒率为22.5%。不同菌株携带dsRNA的大小不尽相同。对6株菌株的dsRNA进行序列测定、组装得到Contigs, Blast比对、分析,鉴定出7种dsRNA病毒和2种+ssRNA病毒,分别与整体病毒科(Totiviridae)、双分病毒科(Partitiviridae)、产黄青霉病毒科(Chrysoviridae)、芜菁黄花叶病毒科(Tymoviridae)和裸露病毒科(Narnaviridae)成员存在序列同源性。【结论】番茄早疫病菌携带 dsRNA存在多样性,不仅携带多种病毒,而且还存在复合侵染现象。已经检测到的9种病毒分属于Totiviridae、Partitiviridae、Tymoviridae、Tymoviridae和Narnaviridae,有些病毒是已经报道的病毒或其新株系,有些则是未被揭示的新病毒。

关键词: 石河子; 番茄早疫病菌; dsRNA; 多样性

Abstract:

【Objective】The objective of this study is to investigate the diversity of dsRNA in A. solani., and to provide new biological control resources for the disease and new understanding of virus diversity and evolutionary.【Methods】Tissue samples with typical symptom were collected from tomato growing areas around Shihezi and pure culture was obtained by tissue isolation and single spore separation. After that, the diversity of dsRNA carried by A. solani. was determined by dsRNA extraction and gel electrophoresis. In the end by RT-PCR using random primers, and molecular cloning, and sequencing techniques, the phylogenetics for the mycovirus carried by 6 strain were analyzed.【Results】The pure culture of 191 strains of A. solani. was obtained and 42 strains with obvious dsRNA bands (from 1 to 4) were found. The mycovirus strains were accounting for 22% of the pure culture strains. The band's sizes were different among the dsRNA carried strains. The Contigs were obtained by sequence and assemble from 6 strains. The blast results showed that the 7 dsRNA virues and 2 +ssRNA virues were shared sequence homology with isolates among the Totiviridae, Partitiviridae, Tymoviridae, Tymoviridae, and(or) Narnaviridae respectively.【Conclusion】There are diverse dsRNAs in A. solani. The strains of A. solani not only carry many mycoviruses, but also mixed infections with two or more unrelated viruses. The 9 mycovirues identified can be divided into 5 known families (Totiviridae, Partitiviridae, Tymoviridae, Tymoviridae, and Narnaviridae). Some are reported viruses or new strains while others are new viruses that have not yet been discovered.

Key words: Shihezi; Alternaria solani; dsRNA; diversity

中图分类号:

S436.412

刘升学, 李思琪, 王晓东, 杨德松. 番茄早疫病菌dsRNA多样性分析[J]. 新疆农业科学, 2023, 60(12): 3057-3064.

LIU Shengxue, LI Siqi, WANG Xiaodong, YANG Desong. Diversity analysis of dsRNA in Alternaria solani[J]. Xinjiang Agricultural Sciences, 2023, 60(12): 3057-3064.

图/表 2

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序号 No.	菌株 Strain	Contigs		已报到病毒Known virus		同源性 Per. Ident (%)	核酸类型 Nucleotide type	分类 Taxonomy
序号 No.	菌株 Strain	组分 Component	大小 Size(nt)	登录号 GenBank	名称/分离物/株系 Name/Isolate/Strain	同源性 Per. Ident (%)	核酸类型 Nucleotide type	分类 Taxonomy
1	AS24	dsRNA-L	2 537	U41345	HvV 190S	63	dsRNA	Totiviridae
				MN935504	BmV1c/CY27	62
				MN935505	BmV1d/CY27	62
				MN935503	BmV1b/CY27	62
				MH396496	BmV1 TMZS01	62
		dsRNA-S	1 483	MT648467	AtPV1	54	dsRNA	Partitiviridae
				MN556985	PVLaPV10 DMG-B_DN51686	53
				MN605502	ENaPV8 PMS8_DN6389	51
2	G3	1	1 189	NC_040793	AaVV1	74	dsRNA	Totiviridae
		1	1 189	KT598230	CmRV CmRV-IL	59	dsRNA	Totiviridae
		2	694	HE579658			dsRNA	Totiviridae
				HE579659	Uncultured totivirus	90
				HE579660
		3	889	HE579658
				HE579659	Uncultured totivirus	100
				HE579660
3	S8	1	3 411	NC_040793	AaVV1	92	dsRNA	Totiviridae
				KT598230	CmRV CmRV-IL	92
				MK758089	BmVV2 CY05	81
		2	559	KJ605397	UvNV-1 GX-10	23	dsRNA	Partitiviridae
4	T4	4	371	MW328751	TpMV1	99	+ssRNA	Tymoviridae
5	H74	1	3 262	KJ817371	AlRV1 HN28	99	dRNA	未分类
6	H63	1	3 684	MK584808	AaCV1		dsRNA	Chrysoviridae
				MN563140	CREA-VE-32SY2 RNA1	98
				NC_040738
				MK672910	AaCV1 QY-2 RNA1	86
		2	2 532	MK584809	AaCV1 CREA-VE-32SY2 RNA2	83
		2	2 532	NC_040739	AaCV1 RNA2	25
		4	1 105	MK584810	AaCV1	97
		4	1 105	NC_040736	CREA-VE-32SY2 RNA3
		5	1 418	MK584811	AaCV1 CREA-VE-32SY2 RNA4	64
		5	1 418	NC_040740	AaCV1 RNA4	99
		3	825	MN605440	ENaNV27 PMS2_317	93	+ssRNA	Narnaviridae

序号 No.	菌株 Strain	Contigs		已报到病毒Known virus		同源性 Per. Ident (%)	核酸类型 Nucleotide type	分类 Taxonomy
序号 No.	菌株 Strain	组分 Component	大小 Size(nt)	登录号 GenBank	名称/分离物/株系 Name/Isolate/Strain	同源性 Per. Ident (%)	核酸类型 Nucleotide type	分类 Taxonomy
1	AS24	dsRNA-L	2 537	U41345	HvV 190S	63	dsRNA	Totiviridae
				MN935504	BmV1c/CY27	62
				MN935505	BmV1d/CY27	62
				MN935503	BmV1b/CY27	62
				MH396496	BmV1 TMZS01	62
		dsRNA-S	1 483	MT648467	AtPV1	54	dsRNA	Partitiviridae
				MN556985	PVLaPV10 DMG-B_DN51686	53
				MN605502	ENaPV8 PMS8_DN6389	51
2	G3	1	1 189	NC_040793	AaVV1	74	dsRNA	Totiviridae
		1	1 189	KT598230	CmRV CmRV-IL	59	dsRNA	Totiviridae
		2	694	HE579658			dsRNA	Totiviridae
				HE579659	Uncultured totivirus	90
				HE579660
		3	889	HE579658
				HE579659	Uncultured totivirus	100
				HE579660
3	S8	1	3 411	NC_040793	AaVV1	92	dsRNA	Totiviridae
				KT598230	CmRV CmRV-IL	92
				MK758089	BmVV2 CY05	81
		2	559	KJ605397	UvNV-1 GX-10	23	dsRNA	Partitiviridae
4	T4	4	371	MW328751	TpMV1	99	+ssRNA	Tymoviridae
5	H74	1	3 262	KJ817371	AlRV1 HN28	99	dRNA	未分类
6	H63	1	3 684	MK584808	AaCV1		dsRNA	Chrysoviridae
				MN563140	CREA-VE-32SY2 RNA1	98
				NC_040738
				MK672910	AaCV1 QY-2 RNA1	86
		2	2 532	MK584809	AaCV1 CREA-VE-32SY2 RNA2	83
		2	2 532	NC_040739	AaCV1 RNA2	25
		4	1 105	MK584810	AaCV1	97
		4	1 105	NC_040736	CREA-VE-32SY2 RNA3
		5	1 418	MK584811	AaCV1 CREA-VE-32SY2 RNA4	64
		5	1 418	NC_040740	AaCV1 RNA4	99
		3	825	MN605440	ENaNV27 PMS2_317	93	+ssRNA	Narnaviridae

番茄早疫病菌dsRNA多样性分析

Diversity analysis of dsRNA in Alternaria solani

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