广东四季海棠叶斑病病原鉴定

doi:10.11924/j.issn.1000-6850.casb2024-0191

中国农学通报 ›› 2025, Vol. 41 ›› Issue (1): 126-131.doi: 10.11924/j.issn.1000-6850.casb2024-0191

广东四季海棠叶斑病病原鉴定

沈会芳(), 杨祁云, 张景欣, 蒲小明, 刘平平, 林壁润, 孙大元()

广东省农业科学院植物保护研究所/广东省植物保护新技术重点实验室，广州 510640

收稿日期:2024-03-18 修回日期:2024-11-08 出版日期:2025-01-05 发布日期:2025-01-01
通讯作者:
孙大元，男，1984年出生，河南信阳人，副研究员，博士，研究方向：作物病虫害防治。通信地址：510640 广东省广州市天河区金颖路7号广东省农业科学院植物保护研究所，E-mail：sundayuan@gdaas.cn。
作者简介:
沈会芳，女，1973年出生，安徽宿州人，研究员，硕士，研究方向：作物病虫害及防控技术研究。通信地址：510640 广东省广州市天河五山金颖路7号广东省农业科学院植物保护研究所，Tel：020-85161048，E-mail：951781658@qq.com。
基金资助:
广东省乡村振兴战略专项-种植业发展类项目“2023年名特优新农产品优质高效低耗生产新技术研究与示范”(2023 ZZ21); 广州市科技计划项目“菜心-玉米-菜心和菜心-水稻-菜心轮作模式的提质增效技术推广应用”(2024E04J1259)

Identification of Pathogen Causing Begonia Leaf Spot in Guangdong

SHEN Huifang(), YANG Qiyun, ZHANG Jingxin, PU Xiaoming, LIU Pingping, LIN Birun, SUN Dayuan()

Institute of Plant Protection, Guangdong Academy of Agricultural Sciences/Key Laboratory of New Technique for Plant Protection in Guangdong, Guangzhou 510640

Received:2024-03-18 Revised:2024-11-08 Published:2025-01-05 Online:2025-01-01

摘要/Abstract

摘要：

为明确广东省四季海棠叶斑病的病原菌种类，从广东省广州市番禺区采集病害样本，采用组织分离法分离病原菌，基于病菌致病性、形态和分子生物学特征确定其分类地位，获得13株形态特征相似的单孢分离物。供试菌株可侵染四季海棠，产生与田间相似的叶斑病症状。在PDA上，菌落浅灰绿色，气生菌丝丰富。分生孢子梗多单生，(36.4~102.8) μm×(4.6~8.6) μm(n=50)。分生孢子褐色，单生，倒棒形、倒梨形、卵形或椭圆形，(25.6~72.5) μm×(15.4~26.3) μm(n=50)，横隔3~7个，纵隔0~3个。基于21株链格孢属内种的ITS、TEF-1α、GAPDH、Alt a1和RPB2基因序列构建系统发育树，病原菌与番茄链格菌的4个菌株单独聚成一支。研究表明，广东四季海棠叶斑病菌为番茄链格孢(Alternaria tomato)，这是番茄链格孢侵染四季海棠的首次报道。

关键词: 四季海棠, 叶斑, 番茄链格孢, 病原鉴定, 分子鉴定

Abstract:

To clarify the pathogen causing leaf spot on Begonia semperflorens in Guangdong, samples of begonia leaf spot from Panyu District, Guangzhou in Guangdong were collected as test materials. Strains were isolated by tissue isolation method. The taxonomic status of pathogens was identified based on pathogenicity determination, morphological and molecular biological characteristics. A total of 13 single-conidium-isolates with similar morphological characteristics were obtained from diseased plant samples. Artificial inoculation results showed that the tested isolates could infect Begonia semperflorens and cause typical leaf spot symptoms observed in the field. Their colonies were light gray-green with abundant aerial hyphae on PDA medium. Conidiophores were brown, solitary, (36.4-102.8) μm×(4.6-8.6) μm (n=50). Conidia were brown, obclavate, obpyriform, oval, (25.6-72.5) μm×(15.4-26.3) μm (n=50), with three to seven transverse and zero to three longitudinal septa. Based on a phylogenetic tree obtained using multiplex alignments of concatenations of ITS, TEF-1α, GAPDH, Alt a1 and RPB2 genes sequences from 21 Alternaria species, two tested isolates were grouped together with four Alternaria tomato strains to form a single clade. This study clarified that the pathogen causing begonia leaf spot in Guangdong was determined as A. tomato, which was reported for the first time that A. tomato could cause begonia leaf spot.

Key words: Begonia semperflorens, leaf spot, Alternaria tomato, pathogen identification, molecular identification

沈会芳, 杨祁云, 张景欣, 蒲小明, 刘平平, 林壁润, 孙大元. 广东四季海棠叶斑病病原鉴定[J]. 中国农学通报, 2025, 41(1): 126-131.

SHEN Huifang, YANG Qiyun, ZHANG Jingxin, PU Xiaoming, LIU Pingping, LIN Birun, SUN Dayuan. Identification of Pathogen Causing Begonia Leaf Spot in Guangdong[J]. Chinese Agricultural Science Bulletin, 2025, 41(1): 126-131.

图/表 6

参考文献 29

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目的基因	正反向引物	引物序列(5'-3')	文献
ITS	ITS1	TCCGTAGGTGAACCTGCGG	[13]
ITS	ITS4	TCCTCCGCTTATTGATATGC	[13]
TEF-1ɑ	TEF1-728F	CATCGAGAAGTTCGAGAAGG	[14]
TEF-1ɑ	TEF1-986R	TACTTGAAGGAACCCTTACC	[14]
GAPDH	GDF1	CAACGGCTTCGGTCGCATTG	[15]
GAPDH	GPD-2	GCCAAGCAGTTGGTTGTGC	[15]
Alt a 1	Alta1F	ATGCAGTTCACCACCATCGC	[16]
Alt a 1	Alta1R	ACGAGGGTGATGTAGGCGTC	[16]
RPB2	RPB2-5F2	GGGGWGAYCAGAAGA AGGC	[17]
RPB2	RPB2-7cR	CCCATRGCTTGTYYRCCC	[17]

目的基因	正反向引物	引物序列(5'-3')	文献
ITS	ITS1	TCCGTAGGTGAACCTGCGG	[13]
ITS	ITS4	TCCTCCGCTTATTGATATGC	[13]
TEF-1ɑ	TEF1-728F	CATCGAGAAGTTCGAGAAGG	[14]
TEF-1ɑ	TEF1-986R	TACTTGAAGGAACCCTTACC	[14]
GAPDH	GDF1	CAACGGCTTCGGTCGCATTG	[15]
GAPDH	GPD-2	GCCAAGCAGTTGGTTGTGC	[15]
Alt a 1	Alta1F	ATGCAGTTCACCACCATCGC	[16]
Alt a 1	Alta1R	ACGAGGGTGATGTAGGCGTC	[16]
RPB2	RPB2-5F2	GGGGWGAYCAGAAGA AGGC	[17]
RPB2	RPB2-7cR	CCCATRGCTTGTYYRCCC	[17]

菌株	ITS	TEF1-α	GAPDH	Alt a 1	RPB2
GBS-4	OR183708	OR192135	OR192139	OR192137	OR192141
GBS-9	OR183687	OR192136	OR192140	OR192138	OR192142
A.a aff. arborescens strain CBS 112633	KP124400	KP125178	KP124252	KP123947	KP124870
A. alstroemeriae strain CBS 118808	KP124296	KP125071	KP124153	KP123845	KP124764
A. alternata strain CBS 795.72	KP124309	KP125085	KP124166	KP123862	KP124778
A. alternata strain CBS 115199	KP124351	KP125127	KP124205	KP123899	KP124819
A. alternata strain CBS 117143	KP124355	KP125131	KP124209	KP123903	KP124823
A. burnsii strain CBS 110.50	KP124421	KP125199	KP124271	KP123968	KP124890
A. burnsii strain CBS 879.95	KP124422	KP125200	KP124272	KP123969	KP124891
A. burnsii strain CBS 118817	KP124424	KP125202	KP124274	KP123971	KP124893
A. gaisen strain CBS 118488	KP124427	KP125206	KP124278	KP123975	KP124897
A. gossypina strain CBS 104.32	KP124430	KP125209	JQ646312	JQ646395	KP124900
A. iridiaustralis strain CBS 118487	KP124436	KP125215	KP124285	KP123982	KP124906
A. jacinthicola strain CBS 133751	KP124438	KP125217	KP124287	KP123984	KP124908
A. jacinthicola strain YZU 171616	MG781011	MG781010	MG781012	MG781013	MG781015
A. longipes strain CBS 121333	KP124444	KP125223	KP124293	KP123990	KP124914
A. senecionicola strain CBS 119545	KP124409	KP125187	KP124260	KP123956	KP124879
A.tenuissima isolate 11	OP476716	OP609768	OP609771	OP609775	OP604538
A. tenuissima isolate 13	OP476718	OP609770	OP609773	OP609776	OP604540
A. tomato strain CBS 103.30	KP124445	KP125224	KP124294	KP123991	KP124915
A. tomato strain CBS 114.35	KP124446	KP125225	KP124295	KP123992	KP124916
A. tomato strain COUFAL0254	MT522410	MT530412	MT530370	MT530384	MT530398
A. tomato strain COUFAL0301	MT522411	MT530413	MT530371	MT530385	MT530399

菌株	ITS	TEF1-α	GAPDH	Alt a 1	RPB2
GBS-4	OR183708	OR192135	OR192139	OR192137	OR192141
GBS-9	OR183687	OR192136	OR192140	OR192138	OR192142
A.a aff. arborescens strain CBS 112633	KP124400	KP125178	KP124252	KP123947	KP124870
A. alstroemeriae strain CBS 118808	KP124296	KP125071	KP124153	KP123845	KP124764
A. alternata strain CBS 795.72	KP124309	KP125085	KP124166	KP123862	KP124778
A. alternata strain CBS 115199	KP124351	KP125127	KP124205	KP123899	KP124819
A. alternata strain CBS 117143	KP124355	KP125131	KP124209	KP123903	KP124823
A. burnsii strain CBS 110.50	KP124421	KP125199	KP124271	KP123968	KP124890
A. burnsii strain CBS 879.95	KP124422	KP125200	KP124272	KP123969	KP124891
A. burnsii strain CBS 118817	KP124424	KP125202	KP124274	KP123971	KP124893
A. gaisen strain CBS 118488	KP124427	KP125206	KP124278	KP123975	KP124897
A. gossypina strain CBS 104.32	KP124430	KP125209	JQ646312	JQ646395	KP124900
A. iridiaustralis strain CBS 118487	KP124436	KP125215	KP124285	KP123982	KP124906
A. jacinthicola strain CBS 133751	KP124438	KP125217	KP124287	KP123984	KP124908
A. jacinthicola strain YZU 171616	MG781011	MG781010	MG781012	MG781013	MG781015
A. longipes strain CBS 121333	KP124444	KP125223	KP124293	KP123990	KP124914
A. senecionicola strain CBS 119545	KP124409	KP125187	KP124260	KP123956	KP124879
A.tenuissima isolate 11	OP476716	OP609768	OP609771	OP609775	OP604538
A. tenuissima isolate 13	OP476718	OP609770	OP609773	OP609776	OP604540
A. tomato strain CBS 103.30	KP124445	KP125224	KP124294	KP123991	KP124915
A. tomato strain CBS 114.35	KP124446	KP125225	KP124295	KP123992	KP124916
A. tomato strain COUFAL0254	MT522410	MT530412	MT530370	MT530384	MT530398
A. tomato strain COUFAL0301	MT522411	MT530413	MT530371	MT530385	MT530399

广东四季海棠叶斑病病原鉴定

Identification of Pathogen Causing Begonia Leaf Spot in Guangdong

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