Identification of Pathogen Causing Begonia Leaf Spot in Guangdong

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

Abstract

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

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.

Figures/Tables 6

References 29

[1]	黄佳茹, 柏晓玉, 程喜悦, 等. NaCl胁迫对四季海棠幼苗生长及膜透性的影响[J]. 吉林农业, 2018(22):42-43.
[2]	容汉诠, 罗蕴芳, 王华荣, 等. 宁夏花卉植物病害名录[J]. 宁夏农学院学报, 1992, 13(4):30-39.
[3]	ZHOU L H, JI G H. First report of bacterial leaf spot of rieger Begonia caused by Xanthomonas campestris pv. begoniae in China[J]. Plant disease, 2013, 97(2):282.
[4]	周丽洪, 王永吉, 韩阳, 等. 丽格海棠细菌性叶斑病病原菌鉴定[J]. 西南师范大学学报(自然科学版), 2012, 37(12):56-61.
[5]	LIN F Y, FANG Z Q, LEE S, et al. First report of Xanthomonas axonopodis pv. begoniae causing bacterial leaf spot on rieger Begonias in Taiwan[J]. Plant disease, 2019, 130(11):2940.
[6]	WU F, LI S C, WANG Y B, et al. First report of Colletotrichum karsti causing anthracnose of Begonia lanternaria in China[J]. Plant disease, 2022, 106(11):2990.
[7]	ZHAI X X, ZHANG B Q, CAO H, et al. First report of Colletotrichum truncatum causing anthracnose of Begonia in China[J]. Plant disease, 2018, 102(6):1177.
[8]	陈玉森. 竹节秋海棠褐斑病原及发病规律观察[J]. 福建热作科技, 2003, 28(3):26.
[9]	BEN H Y, QU H Y, GAO W. First report of Myrothecium roridum causing brown spot disease on hiemalis Begonia in China[J]. Plant disease, 2015, 99(12):1866.
[10]	FUJINAWA M F, PONTES N C, VALE H M, et al. First report of Myrothecium roridum causing Myrothecium leaf spot on Begonia in Brazil[J]. Plant disease, 2016, 100(3):655.
[11]	ZHANG X, JIANG B W, WANG R T, et al. Occurrence of leaf spot of Begonia semperflorens caused by Alternaria tenuissima in China[J]. Plant disease, 2020, 104(8):2292.
[12]	方仲达. 植病研究方法[M]. 北京: 中国农业出版社, 1998:46-50.
[13]	王晋, 王亚茹, 王红宝, 等. 冀中南地区黄冠梨黑斑病病原菌鉴定[J]. 中国果树, 2023(2):68-73.
[14]	CARBONE I, KOHN L M. A method for designing primer sets for speciation studies in filamentous ascomycetes[J]. Mycologia, 1999, 91(3):553-556.
[15]	CARBONE I, KOHN L M. Cochliobolus phylogenetics and the origin of known, highly virulent pathogens, inferred from ITS and glyceraldehyde-3-phosphate dehydrogenase gene sequences[J]. Mycologia, 1999, 91(6):964-977.
[16]	HONG S G, CRAMER R A, LAWRENCE C B, et al. Alt a 1 allergen homologs from Alternaria and related taxa: Analysis of phylogenetic content and secondary structure[J]. Fungal genetics and biology, 2005, 42(2):119-129.
[17]	LIU Y J, WHELEN S, HALL B D. Phylogenetic relationships among ascomycetes: Evidence from an RNA polymerase II subunit[J]. Molecular biology and evolution, 1999, 16:1799-1808.
[18]	张天宇. 中国真菌志(第十六卷:链格孢属)[M]. 北京: 科学出版社, 2003:33-42.
[19]	FREIRE M G, MUSSI-DIAS V, MATTOSO T C, et al. Survey of entophytic Alternaria species isolated from plants in the Brazilian restinga biome[J]. IOSR Journal of pharmacy and biological sciences, 2017, 12(2):84-94.
[20]	石良红, 赵兰勇, 吴迪, 等. 山东牡丹黑斑病的病原菌鉴定与ITS序列分析[J]. 园艺学报, 2015, 42(3):585-590.
[21]	陶航, 扎依娜·玛合巴提, 张烨, 等. 芍药黑斑病病原菌鉴定及对杀菌剂敏感性分析[J]. 园艺学报, 2021, 48(1):173-182. doi: 10.16420/j.issn.0513-353x.2020-0236
[22]	鲁海菊, 潘柳君, 李河, 等. 云南万寿菊叶斑病病原菌的鉴定与ITS序列分析[J]. 西北农业学报, 2015, 24(6):116-119.
[23]	MATIĆ S, TABONE G, GARIBALDI A, et al. Alternaria leaf spot caused by Alternaria species: An emerging problem on ornamental plants in Italy[J]. Plant disease, 2020, 104(8):2275-2287.
[24]	王洪凯, 张天宇. 链格孢属小孢子种的RAPD分析[J]. 菌物系统, 2003, 22(1):35-41.
[25]	WOUDENBERG J H C, GROENEWALD J Z, BINDER M, et al. Alternaria redefined[J]. Studies in mycology, 2013, 75:171-212.
[26]	WOUDENBERG J H C, TRUTER M, GROENEWALD J Z, et al. Large-spored Alternaria pathogens in section Porri disentangled[J]. Studies in mycology, 2014, 79:1-47.
[27]	WOUDENBERG J H C, SEIDL M F, GROENEWALD J Z, et al. Alternaria section Alternaria: Species, formae speciales or pathotypes?[J]. Studies in mycology, 2015, 82:1-21.
[28]	CHANG Y C, YAO R B, CHEN G Y, et al. First report of new guinea Impatiens (Impatiens hawkeri) leaf spot caused by Alternaria burnsii-A. tomato species complex in Taiwan[J]. Plant disease, 2023, 107(7):2251.
[29]	POUDEL B, VALLE V D, HERNÁNDEZ-LAUZARDO A N, et al. First report of Alternaria tomato causing leaf spot on sunflower in Mexico[J]. Plant disease, 2019, 103(5):1029.

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