温郁金根茎腐烂病的病原鉴定

doi:10.3969/j.issn.1004-1524.20230797

浙江农业学报 ›› 2024, Vol. 36 ›› Issue (5): 1086-1093.DOI: 10.3969/j.issn.1004-1524.20230797

温郁金根茎腐烂病的病原鉴定

李亚妮^a(), 陈卫良^a^,^*(), 毛碧增^b^,^*()

浙江大学 a.生物技术研究所,农业农村部作物病虫分子生物学重点实验室,浙江省作物病虫生物学重点实验室; b.新农村发展研究院,浙江杭州 310058

收稿日期:2023-06-27 出版日期:2024-05-25 发布日期:2024-05-29
作者简介:李亚妮(1998—),女,四川广元人,硕士研究生,研究方向为温郁金的病害防治与健康种苗培育。E-mail:1013870281@qq.com
通讯作者: * 陈卫良,E-mail: chenwl@zju.edu.cn;毛碧增,E-mail: maobz@zju.edu.cn
基金资助:
农业农村部农业重大技术协同推广项目(2020XTTGZYC02);浙江省“三农六方”科技协作计划(CTZB-F160728AWZ-SNY1-37)

Pathogen identification of root rot of Curcuma wenyujin

LI Yani^a(), CHEN Weiliang^a^,^*(), MAO Bizeng^b^,^*()

a. Institute of Biotechnology, Key Lab of Molecular Biology of Crop Pathogens and Insects, Ministry of Agriculture and Rural Affairs, Key Laboratory of Biology of Crop Pathogens and Insects of Zhejiang Province; b. The Rural Development Academy, Zhejiang University, Hangzhou 310058, China

Received:2023-06-27 Online:2024-05-25 Published:2024-05-29

摘要/Abstract

摘要：

在浙江省瑞安市陶山镇温郁金主产区,部分田块温郁金发生大规模倒伏、地下根茎严重腐烂。为明确引起该病害的病原物,利用LB培养基和TTC(2,3,5-氯化三苯基四氮唑)培养基从温郁金腐烂根茎中分离获得细菌菌株JF1,采用灌根法、芽侵染法和根茎侵染法确定该菌株对温郁金的致病性。通过形态学特征观察、生理生化测定、多基因序列(16S rDNA、rpoB、gyrB)系统发育分析,明确引起田间温郁金根茎腐烂的病原菌为桑肠杆菌(Enterobacter mori)。

关键词: 温郁金, 根茎腐烂, 病原鉴定, 桑肠杆菌

Abstract:

At main production areas of Curcuma wenyujin in Taoshan Town, Rui’an City, Zhejiang Province, large-scale lodging of C. wenyujin and severe rot of their underground tubers occurred in some fields. To identify the pathogen that causes the disease, the bacteria strain JF1 was isolated from the rotten tuber of C. wenyujin by using LB and TTC (2,3,5-triphenyltetrazolium chloride) medium. The pathogenicity of isolated strain JF1 was confirmed by using three inoculation methods, including root irrigation, bud infection and rhizome infection. The taxonomic status of pathogenic bacteria was identified according to the morphological characteristics, physiological and biochemical tests, phylogenetic analysis of multiple gene sequences (16S rDNA, rpoB, gyrB). The results showed that the pathogenic bacteria causing root rot of C. wenyujin was Enterobacter mori.

Key words: Curcuma wenyujin, root rot, pathogen identification, Enterobacter mori

中图分类号:

S435.672

李亚妮, 陈卫良, 毛碧增. 温郁金根茎腐烂病的病原鉴定[J]. 浙江农业学报, 2024, 36(5): 1086-1093.

LI Yani, CHEN Weiliang, MAO Bizeng. Pathogen identification of root rot of Curcuma wenyujin[J]. Acta Agriculturae Zhejiangensis, 2024, 36(5): 1086-1093.

图/表 7

参考文献 18

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基因 Gene	引物名称 Primer	引物序列 Primer sequence(5'→3')	片段大小 Fragment size/bp
16S rDNA	27F	AGAGTTTGATCCTGGCTCAG	1 500
	1492R	TACGGCTACCTTGTTACGACTT
rpoB	CM7	AACCAGTTCCGCGTTGGCCTGG	1 009
	CM31	CCTGAACAACACGCTCGGA
gyrB	UP-1	GAAGTCATCATGACCGTTCTGCAYGCNGGNGGNAARTTYGA	1 200
	UP-2r	AGCAGGGTACGGATGTGCGAGCCRTCNACRTCNGCRTCNGTCAT

基因 Gene	引物名称 Primer	引物序列 Primer sequence(5'→3')	片段大小 Fragment size/bp
16S rDNA	27F	AGAGTTTGATCCTGGCTCAG	1 500
	1492R	TACGGCTACCTTGTTACGACTT
rpoB	CM7	AACCAGTTCCGCGTTGGCCTGG	1 009
	CM31	CCTGAACAACACGCTCGGA
gyrB	UP-1	GAAGTCATCATGACCGTTCTGCAYGCNGGNGGNAARTTYGA	1 200
	UP-2r	AGCAGGGTACGGATGTGCGAGCCRTCNACRTCNGCRTCNGTCAT

菌株 Strain	GenBank登录号GenBank accession No.
菌株 Strain	16S rDNA	rpoB	gyrB
Enterobacter mori LMG 25706 Enterobacter quasiroggenkampii WCHECL1060	MG846019 KY979139	EU579858 KY979141	JX424992 KY979140
Enterobacter roggenkampii DSM 16690	OP818082	CP017184	CP017184
Enterobacter chengduensis WCHECl-C4	NR179167	KY979144	KY979143
Enterobacter wuhouensis WCHEs120002	MK567957	MK613102	MK613100
Enterobacter chuandaensis 090028	MK049966	MK056243	MK056241
Enterobacter sichuanensis WCHECL1597	MG832788	MG832790	MG832789
Enterobacter bugandensis EB-247	MK603177	LT992502	LT992502
Enterobacter quasihormaechei WCHEs120003	NR180451	MK613106	MK613104
Enterobacter kobei DSM 13645	LT547823	CP017181	CP017181
Enterobacter hormaechei ATCC 49162	OP818074	AJ543724	AY370840
Enterobacter asburiae ATCC 35953	OP818065	AJ543727	CP011863
Enterobacter ludwigii EN-119	MK603179	CP017279	CP017279
Enterobacter cancerogenus LMG 2693	NR116756	AJ566943	FJ617354
Enterobacter oligotrophicus CCA6	OP818079	AP019007	AP019007
Enterobacter cloacae ATCC 13047	NR118568	AJ543726	AB084011
Enterobacter huaxiensis 090008	NR180236	MK056242	MK056248
Enterobacter xiangfangensis 10-17	NR126208	HF679054	CP017183

菌株 Strain	GenBank登录号GenBank accession No.
菌株 Strain	16S rDNA	rpoB	gyrB
Enterobacter mori LMG 25706 Enterobacter quasiroggenkampii WCHECL1060	MG846019 KY979139	EU579858 KY979141	JX424992 KY979140
Enterobacter roggenkampii DSM 16690	OP818082	CP017184	CP017184
Enterobacter chengduensis WCHECl-C4	NR179167	KY979144	KY979143
Enterobacter wuhouensis WCHEs120002	MK567957	MK613102	MK613100
Enterobacter chuandaensis 090028	MK049966	MK056243	MK056241
Enterobacter sichuanensis WCHECL1597	MG832788	MG832790	MG832789
Enterobacter bugandensis EB-247	MK603177	LT992502	LT992502
Enterobacter quasihormaechei WCHEs120003	NR180451	MK613106	MK613104
Enterobacter kobei DSM 13645	LT547823	CP017181	CP017181
Enterobacter hormaechei ATCC 49162	OP818074	AJ543724	AY370840
Enterobacter asburiae ATCC 35953	OP818065	AJ543727	CP011863
Enterobacter ludwigii EN-119	MK603179	CP017279	CP017279
Enterobacter cancerogenus LMG 2693	NR116756	AJ566943	FJ617354
Enterobacter oligotrophicus CCA6	OP818079	AP019007	AP019007
Enterobacter cloacae ATCC 13047	NR118568	AJ543726	AB084011
Enterobacter huaxiensis 090008	NR180236	MK056242	MK056248
Enterobacter xiangfangensis 10-17	NR126208	HF679054	CP017183

编号 Number	特性 Property	结果 Result	分离菌株JF1 Isolated strain JF1	模式菌株E. mori Type strain E. mori	阴性对照 Negative control
1	流动性Motility	穿刺线周围呈雾状Foggy puncture line	+	+	-
2	鸟氨酸脱羧酶Ornithine decarboxylase	紫色Purple	+	+	-
3	赖氨酸脱羧酶Lysine decarboxylase	黄色Yellow	-	-	-
4	甲基红试验Methyl red test	不变色Unchanged	-	-	-
5	V-P试验Voges-Proskauer test	红色Red	+	+	-
6	西蒙氏枸橼酸盐Simmons citrate	蓝色Blue	+	+	-
7	硫化氢生成H₂S production	不变色Unchanged	-	-	-
8	尿素酶Urease	黄色Yellow	-	-	-
9	色氨酸Tryptophan	不变色Unchanged	-	-	-
10	苯丙氨酸Phenylalanine	不变色Unchanged	-	-	-
11	甘露醇Mannitol	黄色Yellow	+	+	-
12	肌醇Inositol	绿色Green	-	-	-
13	山梨醇Sorbitol	黄色Yellow	+	+	-
14	蜜二糖Melibiose	黄色Yellow	+	+	-
15	核糖醇Ribitol	绿色Green	-	-	-
16	棉子糖Raffinose	黄色Yellow	+	+	-

温郁金根茎腐烂病的病原鉴定

Pathogen identification of root rot of Curcuma wenyujin

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