Isolation and identification of the pathogen causing tomato canker in Zhejiang Province of China

doi:10.3969/j.issn.1004-1524.20240240

Acta Agriculturae Zhejiangensis ›› 2025, Vol. 37 ›› Issue (6): 1293-1300.DOI: 10.3969/j.issn.1004-1524.20240240

• Plant Protection • Previous Articles Next Articles

Isolation and identification of the pathogen causing tomato canker in Zhejiang Province of China

LIU Pengfei¹^,²(), ZHANG Shuhan¹^,², HONG Kai¹^,², SHAO Yue³, LOU Binggan¹^,²^,^*()

1. Institute of Biotechnology, Zhejiang University, Hangzhou 310058, China
2. Key Laboratory of Molecular Biology of Crop Pathogens and Insects, Ministry of Agriculture and Rural Affairs, Hangzhou 310058, China
3. The Plant Protection and Soil Fertilizer Management Station of Wenzhou, Wenzhou 325088, Zhejiang, China

Received:2024-03-15 Online:2025-06-25 Published:2025-07-08

Abstract

Abstract:

Tomato bacterial canker, a devastating bacterial disease affecting tomatoes, has not been previously reported in Zhejiang Province. This study focused on the first occurrence of tomato bacterial canker in Zhejiang, conducting pathogen isolation, purification, morphological identification, and pathogenicity testing. Seven pathogenic strains were isolated, exhibiting yellow circular smooth colonies on 523 medium. The bacterial cells were rod-shaped, measuring (0.40-0.75) μm×(1.20-3.30) μm, and non-flagellated. Inoculation caused leaf curling and desiccation, stem cankers, and bird’s eye spots on tomato fruits. Further sequencing, comparison, and phylogenetic analysis of 16S rRNA and six housekeeping genes revealed that all seven pathogenic strains showed high consistency with Clavibacter michiganensis subsp. michiganensis, the causal agent of tomato bacterial canker. Comprehensive morphological, pathogenic, and molecular identification results demonstrated that the newly emerged disease on tomatoes in Wenzhou, Zhejiang, is tomato bacterial canker.

Key words: tomato bacterial canker, pathogen identification, Clavibacter michiganensis subsp. michiganensis

CLC Number:

S436.412

LIU Pengfei, ZHANG Shuhan, HONG Kai, SHAO Yue, LOU Binggan. Isolation and identification of the pathogen causing tomato canker in Zhejiang Province of China[J]. Acta Agriculturae Zhejiangensis, 2025, 37(6): 1293-1300.

Figures/Tables 6

References 26

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基因 Gene	引物序列 Primer sequence	片段大小 Length/ bp	参考文献 Reference
16S rRNA	F:5'-CGGTCTACAACGCCCTCAAGA-3' R:5'- TGCGTGAAGCGGAAGATGTTG-3'	1 465	[9]
atpD	F:5'-CGGTCTACAACGCCCTCAAGA-3' R:5'-TGCGTGAAGCGGAAGATGTTG-3'	697	[10]
kdpA	F:5'-GTGCAGAACTTCGTCTCGG-3' R:5'-GAGCATCATGTTGATCATCG-3'	693	[11]
ppK	F:5'-GAGAACCTCATCCAGGCCCT-3' R:5'-GAGAACCTCATCCAGGCCCT-3'	604	[10]
sdhA	F:5'-CCTGGATGTTCGTGTACC-3' R:5'-GAGGACATGGAGTTCTTCC-3'	778	[11]
recA	F:5'-TCGGCAAGGGCTCGGTCATGC-3' R:5'-GGTCGCCRTCGTASGTGTACCA-3'	830	[12]
rpoB	F:5'-CATCATCAACGGCACCGAGC-3' R:5'-AAGCCGAAGGGGTTGATGCG-3'	1 037	[12]

基因 Gene	引物序列 Primer sequence	片段大小 Length/ bp	参考文献 Reference
16S rRNA	F:5'-CGGTCTACAACGCCCTCAAGA-3' R:5'- TGCGTGAAGCGGAAGATGTTG-3'	1 465	[9]
atpD	F:5'-CGGTCTACAACGCCCTCAAGA-3' R:5'-TGCGTGAAGCGGAAGATGTTG-3'	697	[10]
kdpA	F:5'-GTGCAGAACTTCGTCTCGG-3' R:5'-GAGCATCATGTTGATCATCG-3'	693	[11]
ppK	F:5'-GAGAACCTCATCCAGGCCCT-3' R:5'-GAGAACCTCATCCAGGCCCT-3'	604	[10]
sdhA	F:5'-CCTGGATGTTCGTGTACC-3' R:5'-GAGGACATGGAGTTCTTCC-3'	778	[11]
recA	F:5'-TCGGCAAGGGCTCGGTCATGC-3' R:5'-GGTCGCCRTCGTASGTGTACCA-3'	830	[12]
rpoB	F:5'-CATCATCAACGGCACCGAGC-3' R:5'-AAGCCGAAGGGGTTGATGCG-3'	1 037	[12]

种名 Species name	菌株 Strain	来源 Origin
Clavibacter michiganensis subsp. michiganensis	CN0102	浙江温州(本研究) Wenzhou, Zhejiang, China
C. m. subsp. michiganensis	NCPPB2979	匈牙利Hungary
C. m. subsp. michiganensis	ATCC4450	未知Unknow
C. m. subsp. michiganensis	7291-3	甘肃酒泉 Jiuquan, Gansu, China
C. m. subsp. michiganensis	3315-3	中国China
C. m. subsp. michiganensis	UF1	美国America
C. m. subsp. michiganensis	VL527	智利Chile
C. m. subsp. michiganensis	A5747*	美国America
C. m. subsp. michiganensis	N.P.	白俄罗斯
		Republic of Belarus
C. m. subsp. michiganensis	A4758	中国China
C. m. subsp. sepedonicus	K496	美国America
C. m. subsp. insidiosus	R1-1	美国America
C. m. subsp. nebraskensis	CNK-2	美国America
C. m. subsp. tessellarius	ATCC33566	美国America
Ralstonia solanacearum	UW251	哥伦比亚Columbia

种名 Species name	菌株 Strain	来源 Origin
Clavibacter michiganensis subsp. michiganensis	CN0102	浙江温州(本研究) Wenzhou, Zhejiang, China
C. m. subsp. michiganensis	NCPPB2979	匈牙利Hungary
C. m. subsp. michiganensis	ATCC4450	未知Unknow
C. m. subsp. michiganensis	7291-3	甘肃酒泉 Jiuquan, Gansu, China
C. m. subsp. michiganensis	3315-3	中国China
C. m. subsp. michiganensis	UF1	美国America
C. m. subsp. michiganensis	VL527	智利Chile
C. m. subsp. michiganensis	A5747*	美国America
C. m. subsp. michiganensis	N.P.	白俄罗斯
		Republic of Belarus
C. m. subsp. michiganensis	A4758	中国China
C. m. subsp. sepedonicus	K496	美国America
C. m. subsp. insidiosus	R1-1	美国America
C. m. subsp. nebraskensis	CNK-2	美国America
C. m. subsp. tessellarius	ATCC33566	美国America
Ralstonia solanacearum	UW251	哥伦比亚Columbia

Isolation and identification of the pathogen causing tomato canker in Zhejiang Province of China

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