福建省番茄青枯病菌遗传多样性分析及其抗性砧木的筛选

doi:10.16420/j.issn.0513-353x.2024-0395

园艺学报 ›› 2024, Vol. 51 ›› Issue (11): 2540-2554.doi: 10.16420/j.issn.0513-353x.2024-0395

• 遗传育种·种质资源·分子生物学 • 上一篇下一篇

福建省番茄青枯病菌遗传多样性分析及其抗性砧木的筛选

王荣波¹, 王海云², 张前荣³, 蔡松龄¹, 李本金¹, 张美祥⁴, 刘裴清¹^,^*()

¹ 福建省农业科学院植物保护研究所，福建省作物有害生物监测与治理重点实验室，福州 350013
² 福建省福州市永泰县种植业服务中心，福州 350799
³ 福建省农业科学院作物研究所，福州 350013
⁴ 陕西师范大学生命科学学院，西安 710119

收稿日期:2024-05-28 修回日期:2024-09-08 出版日期:2024-12-12 发布日期:2024-11-25
通讯作者:
^* E-mail：liupeiqing11@163.com
基金资助:
国家自然科学基金项目(32072400); 福建省农业科学院“5511”协同创新工程项目(XTCXGC2021011); 福建省农业科学院“5511”协同创新工程项目(XTCXGC2021017); 厦门市重大科技项目(3502Z20221018); 福建省农业科学院国家自然科学基金延伸研究项目(GJYS202204); 福建省省属公益类竞争性项目(2023R1070); 福建省省属公益类竞争性项目(2023R1072)

Genetic Diversity of Ralstonia solanacearum Strains Isolated from Fujian Province and Evaluation of Tomato Rootstocks for Resistance to Bacterial Wilt

WANG Rongbo¹, WANG Haiyun², ZHANG Qianrong³, CAI Songling¹, LI Benjin¹, ZHANG Meixiang⁴, LIU Peiqing¹^,^*()

¹ Fujian Key Laboratory for Monitoring and Integrated Management of Crop Pests，Institute of Plant Protection，Fujian Academy of Agricultural Sciences，Fuzhou 350013，China
² Yongtai County Crop Cultivation Service Center，Fuzhou 350799，China
³ Crops Research Institute，Fujian Academy of Agricultural Sciences，Fuzhou 350013，China
⁴ College of Life Sciences，Shaanxi Normal University，Xi’an 710119，China

Received:2024-05-28 Revised:2024-09-08 Published:2024-12-12 Online:2024-11-25

摘要/Abstract

摘要：

2021—2022年，从中国福建省5个主要番茄种植区分离得到62株青枯病菌株（Ralstonia solanacearum）。通过演化型特异性多重聚合酶链式反应（phylotype-specific multiplex polymerase chain reaction，Pmx-PCR）、生理小种和生化型的鉴定，发现所有菌株均属于演化型Ⅰ和生理小种1，其中61株为生化型Ⅲ，1株为非典型生化型。基于egl基因部分序列的系统发育分析进一步将演化型Ⅰ细分为序列变种14、15、16、17和34，其中序列变种15是主要优势变种。对13个番茄砧木品种进行了青枯菌抗性评估，结果显示，不同砧木品种对不同序列变种的青枯菌表现出明显的抗性差异，‘桂砧1号’和‘威尔神根’对福建省的优势菌系表现出广谱抗性。

关键词: 番茄, 青枯劳尔氏菌, 遗传多样性, 砧木, 抗病性

Abstract:

In this study，62 strains of Ralstonia solanacearum were isolated in 2021 to 2022 from five tomato-growing areas in Fujian，China. Using phylotype-specific multiplex PCR（Pmx-PCR），the biochemical and race tests，all strains were identified as phylotype I and race 1. Of these，61 strains were identified as belonging to biovar Ⅲ，while one strain was classified as atypical biovar. The strains were further divided five different sequevars（14，15，16，17，and 34）based on polymorphisms in the endoglucanase（egl）gene. Among the five sequevars，sequevar 15 was the predominant type in Fujian Province. In addition，the resistance of 13 tomato rootstock cultivars to R. solanacearum was evaluated，revealing significant differences in resistance among the cultivars to different sequence types of the pathogen. It is noteworthy that two rootstock cultivars，‘Guizhen 1’and‘Weier Shengen’，exhibited broad-spectrum resistance to the predominant strains in Fujian Province.

Key words: tomato, Ralstonia solanacearum, genetic diversity, rootstock, disease resistance

王荣波, 王海云, 张前荣, 蔡松龄, 李本金, 张美祥, 刘裴清. 福建省番茄青枯病菌遗传多样性分析及其抗性砧木的筛选[J]. 园艺学报, 2024, 51(11): 2540-2554.

WANG Rongbo, WANG Haiyun, ZHANG Qianrong, CAI Songling, LI Benjin, ZHANG Meixiang, LIU Peiqing. Genetic Diversity of Ralstonia solanacearum Strains Isolated from Fujian Province and Evaluation of Tomato Rootstocks for Resistance to Bacterial Wilt[J]. Acta Horticulturae Sinica, 2024, 51(11): 2540-2554.

图/表 12

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用途 Application	材料 Material	来源 Source
番茄砧木 Tomato rootstock	桂砧1号 Guizhen 1	广西南宁赛绿农业科技有限公司 Guangxi Nanning Sailü Agricultural Technology Co., Ltd.
	丽砧1号 Lizhen 1	大连市北蔬农业有限公司 Dalian Beishu Agriculture Co.，Ltd.
	强力 Qiangli	山东寿光市春秋种业总经销 Shandong Shouguang Spring and Autumn Seed Industry general distributor
	朗达240 Langda 240	北京君川种业科技有限公司 Beijing Junchuan Seed Technology Co.，Ltd.
	威尔神根 Weier Shengen	山东威尔种子有限公司 Shandong Weier Seed Co.，Ltd.
	番茄砧木6号 Tomato rootstock 6	以色列尼瑞特种业有限公司 Israel Nirait Seed Industry Co.，Ltd.
	番茄砧木405 Tomato rootstock 405	上海惠和种业有限公司 Shanghai Huihe Seed Industry Co.，Ltd.
	乾德番砧 Qiande tomato rootstock	上海乾德种业有限公司 Shanghai Qiande Seed Industry Co.，Ltd.
	浙砧1号 Zhezhen 1	浙江依农种业有限公司 Zhejiang Yinong Seed Industry Co.，Ltd.
番茄、茄子两用砧木 Tomato and eggplant rootstock	西茄砧木Xiqie rootstock	北京络芙特科贸有限公司 Beijing Luofute Science and Trade Co.，Ltd.
番茄、茄子两用砧木 Tomato and eggplant rootstock	亚拉 Yala	贵州络芙特生物科技有限公司 Guizhou Lofute Biotechnology Co.，Ltd.
多用砧木Multipurpose rootstock	大力神砧木F₁ Dalishen rootstock F₁	寿光市宏伟种业有限公司 Shouguang Hongwei Seed Industry Co.，Ltd.
茄子砧木 Eggplant rootstock	京丰808 Jingfeng 808	山东威尔种子有限公司 Shandong Weier Seed Co.，Ltd.

用途 Application	材料 Material	来源 Source
番茄砧木 Tomato rootstock	桂砧1号 Guizhen 1	广西南宁赛绿农业科技有限公司 Guangxi Nanning Sailü Agricultural Technology Co., Ltd.
	丽砧1号 Lizhen 1	大连市北蔬农业有限公司 Dalian Beishu Agriculture Co.，Ltd.
	强力 Qiangli	山东寿光市春秋种业总经销 Shandong Shouguang Spring and Autumn Seed Industry general distributor
	朗达240 Langda 240	北京君川种业科技有限公司 Beijing Junchuan Seed Technology Co.，Ltd.
	威尔神根 Weier Shengen	山东威尔种子有限公司 Shandong Weier Seed Co.，Ltd.
	番茄砧木6号 Tomato rootstock 6	以色列尼瑞特种业有限公司 Israel Nirait Seed Industry Co.，Ltd.
	番茄砧木405 Tomato rootstock 405	上海惠和种业有限公司 Shanghai Huihe Seed Industry Co.，Ltd.
	乾德番砧 Qiande tomato rootstock	上海乾德种业有限公司 Shanghai Qiande Seed Industry Co.，Ltd.
	浙砧1号 Zhezhen 1	浙江依农种业有限公司 Zhejiang Yinong Seed Industry Co.，Ltd.
番茄、茄子两用砧木 Tomato and eggplant rootstock	西茄砧木Xiqie rootstock	北京络芙特科贸有限公司 Beijing Luofute Science and Trade Co.，Ltd.
番茄、茄子两用砧木 Tomato and eggplant rootstock	亚拉 Yala	贵州络芙特生物科技有限公司 Guizhou Lofute Biotechnology Co.，Ltd.
多用砧木Multipurpose rootstock	大力神砧木F₁ Dalishen rootstock F₁	寿光市宏伟种业有限公司 Shouguang Hongwei Seed Industry Co.，Ltd.
茄子砧木 Eggplant rootstock	京丰808 Jingfeng 808	山东威尔种子有限公司 Shandong Weier Seed Co.，Ltd.

鉴定用途 Application	引物名称 Primer name	引物序列（5′-3′） Primer sequence	大小/bp Size
青枯菌 Ralstonia solanacearum	AU759	GTCGCCGTCAACTCACTTTCC	281
青枯菌 Ralstonia solanacearum	AU760	GTCGCCGTCAGCAATGCGGAATCG
演化型Ⅰ上游 Phylotype I-F	Nmult21:1F	CGTTGATGAGGCGCGCAATTT	144
演化型Ⅱ上游 Phylotype Ⅱ-F	Nmult21:2F	AAGTTATGGACGGTGGAAGTC	372
演化型Ⅲ上游 Phylotype Ⅲ-F	Nmult23:AF	ATTACAGAGCAATCGAAAGATT	91
演化型Ⅳ上游 Phylotype Ⅳ-F	Nmult22:InF	ATTGCCAAGACGAGAGAAGTA	213
演化型下游 Phylotype-R	Nmult22:RR	TCGCTTGACCCTATAACGAGTA
生理小种1 Race 1	PS-IS	F：CGCAACGCTGGATGAACCC；R：CAGACGATGCGAAGCCTGAC	1 070
生理小种2 Race 2	ISRso19	F：TGGGAGAGGATGGCGGCTTT；R：TGACCCGCCTTTCGGTGTTT	1 884
生理小种4 Race 4	21	F：CGACGCTGACGAAGGGACTC；R：CTGACACGGCAAGCGCTCA	125
egl	Endo	F：ATGCATGCCGCTGGTCGCCGC；R：GCGTTGCCCGGCACGAACACC	698

鉴定用途 Application	引物名称 Primer name	引物序列（5′-3′） Primer sequence	大小/bp Size
青枯菌 Ralstonia solanacearum	AU759	GTCGCCGTCAACTCACTTTCC	281
青枯菌 Ralstonia solanacearum	AU760	GTCGCCGTCAGCAATGCGGAATCG
演化型Ⅰ上游 Phylotype I-F	Nmult21:1F	CGTTGATGAGGCGCGCAATTT	144
演化型Ⅱ上游 Phylotype Ⅱ-F	Nmult21:2F	AAGTTATGGACGGTGGAAGTC	372
演化型Ⅲ上游 Phylotype Ⅲ-F	Nmult23:AF	ATTACAGAGCAATCGAAAGATT	91
演化型Ⅳ上游 Phylotype Ⅳ-F	Nmult22:InF	ATTGCCAAGACGAGAGAAGTA	213
演化型下游 Phylotype-R	Nmult22:RR	TCGCTTGACCCTATAACGAGTA
生理小种1 Race 1	PS-IS	F：CGCAACGCTGGATGAACCC；R：CAGACGATGCGAAGCCTGAC	1 070
生理小种2 Race 2	ISRso19	F：TGGGAGAGGATGGCGGCTTT；R：TGACCCGCCTTTCGGTGTTT	1 884
生理小种4 Race 4	21	F：CGACGCTGACGAAGGGACTC；R：CTGACACGGCAAGCGCTCA	125
egl	Endo	F：ATGCATGCCGCTGGTCGCCGC；R：GCGTTGCCCGGCACGAACACC	698

生化型 Biovar	乳糖 Lactose	麦芽糖 Maltose	纤维二糖 Cellobiose	甘露醇 Mannitol	山梨醇 Sorbitol	甜醇 Dulcitol
Ⅰ	-	-	-	-	-	-
Ⅱ	+	+	+	-	-	-
Ⅲ	+	+	+	+	+	+
Ⅲ-1	+	+	+	+	+	-
Ⅲ-2	+	-	+	+	+	-
Ⅳ	-	-	-	+	+	+
Ⅴ	+	+	+	+	-	-

福建省番茄青枯病菌遗传多样性分析及其抗性砧木的筛选

Genetic Diversity of Ralstonia solanacearum Strains Isolated from Fujian Province and Evaluation of Tomato Rootstocks for Resistance to Bacterial Wilt

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参考菌株 Reference strain	寄主 Host	来源 Origin	演化型 Phylotype	序列变种 Sequevar	egl登录号 Accession number
R288	桑树Morus alba L.	中国 China	Ⅰ	12	GQ907153
JT523	马铃薯 Solanum tuberosum L.	留尼汪岛 Reunion Island	Ⅰ	13	AF295252
PSS81	番茄 Solanum lycopersicum L.	中国 China	Ⅰ	14	FJ561066
PSS358	番茄 Solanum lycopersicum L.	中国 China	Ⅰ	15	EU407298
UW151	姜 Zingiber officinale Roscoe	澳大利亚 Australia	Ⅰ	16	AF295254
P11	花生 Arachis hypogaea L.	中国 China	Ⅰ	17	FJ561068
GMI1000	番茄 Solanum lycopersicum L.	法国 France	Ⅰ	18	AF295251
JT519	天竺葵 Pelargonium hortorum Bailey	留尼汪岛 Reunion Island	Ⅰ	31	GU295032
PSS219	番茄 Solanum lycopersicum L.	中国 China	Ⅰ	34	FJ561167
O3	橄榄树 Canarium album Rauesch	中国 China	Ⅰ	44	FJ561069
Tb28	烟草 Nicotiana tabacum L.	中国 China	Ⅰ	44	FJ561127
Tb43	烟草 Nicotiana tabacum L.	中国 China	Ⅰ	44	FJ561129
Bd11	木槿 Hibiscus syriacus L.	中国 China	Ⅰ	44	FJ561098
CIP365	马铃薯 Solanum tuberosum L.	菲律宾 Philippines	Ⅰ	45	GQ907151
MAD17	辣椒 Capsicum annuum L.	马达加斯加 Madagascar	Ⅰ	46	GU295040
GMI8254	番茄 Solanum lycopersicum L.	印度尼西亚 Indonesia	Ⅰ	47	GU295014
M2	桑树 Morus alba L.	中国 China	Ⅰ	48	FJ561067
CMR87	番茄 Solanum lycopersicum L.	喀麦隆 Cameroon	Ⅱ	35	EF439727
CMR121	番茄 Solanum lycopersicum L.	喀麦隆 Cameroon	Ⅱ	52	EF439725
CMR39	番茄 Solanum lycopersicum L.	喀麦隆 Cameroon	Ⅱ	41	EF439726
CFBP2972	马铃薯 Solanum tuberosum L.	马提尼克 Martinique	Ⅱ	35	EF371809
UW551	天竺葵 Pelargonium hortorum Bailey	肯尼亚 Kenya	Ⅱ	1	DQ657596
ICMP7963	马铃薯 Solanum tuberosum L.	肯尼亚 Kenya	Ⅱ	7	AF295263
UW162	香蕉 Musa nana Lour.	秘鲁 Peru	Ⅱ	4	AF295256
MOLK2	香蕉 Musa nana Lour.	菲律宾Philippines	Ⅱ	3	EF371841
CMR66	木龙葵 Solanum scabrum Mill.	喀麦隆Cameroon	Ⅲ	49	EF439729
JT525	天竺葵 Pelargonium hortorum Bailey	留尼汪岛 Reunion Island	Ⅲ	19	AF295272
CFBP3059	茄子 Solanum melongena L.	布基纳法索The Burkina Faso	Ⅲ	23	AF295270
NCPPB332	马铃薯 Solanum tuberosum L.	津巴布韦 Zimbabwe	Ⅲ	22	DQ657649
MAFF301558	马铃薯 Solanum tuberosum L.	日本 Japan	Ⅳ	8	AY465002
Psi7	番茄 Solanum lycopersicum L.	印度尼西亚 Indonesia	Ⅳ	10	EF371804
ACH732	番茄 Solanum lycopersicum L.	澳大利亚 Australia	Ⅳ	11	GQ907150

地点 Origin	菌株 Strain	生理小种 Race	演化型 Phylotype	序列变种 Sequevar	生化型 Biovar	GenBank序列号 GenBank accession number
福州市	RS210523F	1	Ⅰ	14	Ⅲ	PP639179
Fuzhou	RS210527F	1	Ⅰ	14	Ⅲ	PP639177
	RS210531F	1	Ⅰ	14	Ⅲ	PP639178
	RS20220502F	1	Ⅰ	15	Ⅲ	PP639152
	RS20220503F	1	Ⅰ	15	Ⅲ	PP639153
	RS20220504F	1	Ⅰ	15	Ⅲ	PP639154
	RS20220505FT	1	Ⅰ	15	Ⅲ	PP639155
	RS20220501F	1	Ⅰ	15	Ⅲ	PP639151
	RS210507F	1	Ⅰ	34	Ⅲ	PP639142
	RS210553FT	1	Ⅰ	34	Ⅲ	PP639143
	RS210557F	1	Ⅰ	34	Ⅲ	PP639144
	RS210561FT	1	Ⅰ	34	Ⅲ	PP639171
宁德市	RS202107129F	1	Ⅰ	14	Ⅲ	PP639182
Ningde	RS202107131F	1	Ⅰ	14	Ⅲ	PP639183
	RS202107132F	1	Ⅰ	14	Ⅲ	PP639184
	RS2107128F	1	Ⅰ	14	Ⅲ	PP639167
	RS2107130F	1	Ⅰ	14	Ⅲ	PP639168
	RS2107133F	1	Ⅰ	14	Ⅲ	PP639169
	RS2107139F	1	Ⅰ	15	Ⅲ	PP639166
	RS202107134F	1	Ⅰ	17	Ⅲ	PP639185
	RS202107137F	1	Ⅰ	17	Ⅲ	PP639186
	RS2107135F	1	Ⅰ	17	Ⅲ	PP639170
	RS2107136F	1	Ⅰ	17	Ⅲ	PP639164
	RS2107138F	1	Ⅰ	17	Ⅲ	PP639165
莆田市	RS210599FT	1	Ⅰ	14	Ⅲ	PP639173
Putian	RS2105107FT	1	Ⅰ	15	Ⅲ	PP639149
	RS2105115FT	1	Ⅰ	15	Ⅲ	PP639150
	RS210564F	1	Ⅰ	15	Ⅲ	PP639145
	RS210566F	1	Ⅰ	15	Ⅲ	PP639187
	RS210571F	1	Ⅰ	15	Ⅲ	PP639188
	RS210584F	1	Ⅰ	15	Ⅲ	PP639147
	RS210586F	1	Ⅰ	15	Ⅲ	PP639189
	RS210592F	1	Ⅰ	15	Ⅲ	PP639148
	RS210597F	1	Ⅰ	15	Ⅲ	PP639190
	RS210576F	1	Ⅰ	16	Ⅳ	PP639146
	RS2105108FT	1	Ⅰ	17	Ⅲ	PP639176
	RS210568F	1	Ⅰ	34	Ⅲ	PP639172
三明市	RS210422F	1	Ⅰ	15	Ⅲ	PP639180
Sanming	RS210437F	1	Ⅰ	16	Ⅲ	PP639198
	RS210438F	1	Ⅰ	16	Ⅲ	PP639175
	RS210439F	1	Ⅰ	16	Ⅲ	PP639199
	RS210417F	1	Ⅰ	17	Ⅲ	PP639191
	RS210418F	1	Ⅰ	17	Ⅲ	PP639192
	RS210420F	1	Ⅰ	17	Ⅲ	PP639193
	RS210424F	1	Ⅰ	17	Ⅲ	PP639194
	RS210426F	1	Ⅰ	17	Ⅲ	PP639181
	RS210428F	1	Ⅰ	17	Ⅲ	PP639174
	RS210431F	1	Ⅰ	17	Ⅲ	PP639195
	RS210433F	1	Ⅰ	17	Ⅲ	PP639196
	RS210435F	1	Ⅰ	17	Ⅲ	PP639197
漳州市	RS211205F	1	Ⅰ	15	Ⅲ	PP639158
Zhangzhou	RS2112115FT	1	Ⅰ	15	Ⅲ	PP639159
	RS2112124FT	1	Ⅰ	15	Ⅲ	PP639160
	RS211215F	1	Ⅰ	15	Ⅲ	PP639161
	RS211235F	1	Ⅰ	15	Ⅲ	PP639162
	RS211242F	1	Ⅰ	15	Ⅲ	PP639163
	RS211101F	1	Ⅰ	16	Ⅲ	PP639156
	RS211103F	1	Ⅰ	16	Ⅲ	PP639157
	RS202112112FT	1	Ⅰ	17	Ⅲ	PP639202
	RS202112121FT	1	Ⅰ	34	Ⅲ	PP639203
	RS20211219F	1	Ⅰ	34	Ⅲ	PP639200
	RS20211229F	1	Ⅰ	34	Ⅲ	PP639201

砧木 Rootstock	RS210507F （序列变种34 Sequevar 34）	RS2107128F （序列变种14 Sequevar 14）	RS210564F （序列变种15 Sequevar 15）	RS210426F （序列变种17 Sequevar 17）	RS211101F （序列变种16 Sequevar 16）	GMI1000 （对照，序列变种18 Control，sequevar 18）
小汤姆 MicroTom	S	S	MS	S	S	S
桂砧1号 Guizhen 1	R	R	MR	MR	R	MR
丽砧1号 Lizhen 1	MS	S	MS	MR	MR	MR
强力 Qiangli	S	S	MS	MS	MS	MR
朗达240 Langda 240	S	MS	S	S	S	S
威尔神根 Weier Shengen	R	MS	R	R	R	R
番茄砧木6号 Tomato rootstock 6	MS	MS	MR	MS	MS	MR
番茄砧木405 Tomato rootstock 405	S	S	MR	MR	R	MR
乾德番砧 Qiande tomato rootstock	MR	MS	MS	MS	MS	MS
浙砧1号 Zhezhen 1	MS	MR	MR	MS	MS	S
西茄砧木 Xiqie rootstock	MS	MS	MR	MR	R	MS
亚拉 Yala	MS	MS	MR	MR	S	MS
大力神砧木F₁Dalishen rootstock F₁	MS	S	S	MS	S	S
京丰808 Jingfeng 808	HS	HS	S	S	S	HS

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福建省番茄青枯病菌遗传多样性分析及其抗性砧木的筛选

Genetic Diversity of Ralstonia solanacearum Strains Isolated from Fujian Province and Evaluation of Tomato Rootstocks for Resistance to Bacterial Wilt

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