贵州省白及叶褐斑病病原菌鉴定及其生物学特性
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摘要
为明确贵州省白及叶褐斑病病原菌及其生物学特性,采用组织分离法和离体接种法分别对其病原菌进行分离和致病性测定,利用形态学及ITS、EF-1α、β-tubulin基因序列分析对病原菌的分类进行鉴定,并对病原菌的生物学特性进行研究。结果表明:分离得到的白及叶褐斑病病原菌菌株编号为HGUP_(17)355;该菌菌丝白色、羊绒状,菌落中心隆起呈淡黄色、背面枫叶红色,大型分生孢子窄镰刀形,3～8个分隔,未见厚垣孢子,小型分生孢子卵圆形,1～2个分隔;在以ITS、EF-1α、β-tubulin三个基因序列构建的系统发育树中,菌株HGUP_(17)355与燕麦镰刀菌Fusarium avenaceum聚于一支,且支持率达100%,结合形态特征与分子生物学将其鉴定为燕麦镰刀菌。该菌菌丝最适生长温度为20℃,最适生长pH为6,在燕麦片琼脂培养基上生长最快,以麦芽糖为碳源、以L-苯丙氨酸为氮源时利用率最高,致死温度为45℃,对光照不敏感。
To clarify the pathogen causing leaf brown spots of Chinese ground orchid Bletilla striata in Guizhou Province and its biological characteristics, the pathogen was obtained by using the method of tissue isolation. Its pathogenicity was tested in vitro inoculated leaves, and the pathogen was classified and identified with morphology and phylogeny based on combined ITS, EF-1α and β-tubulin sequence analysis; the biological characteristics of pathogen were also studied. The results showed that the strain isolated from leaf brown spots of B. striata was named as HGUP17355. Its main diagnostic features were as followed: white mycelia, cashmere-like form, uplifted center of colonies, faint yellow, maple-leaf red dorsal surface, oval microconidia with one-two septa, narrow sickle-like macroconidia with three-eight septa, and no chlamydospores. The phylogenetic tree based on combined ITS, EF-1α, β-tubulin sequences showed that HGUP17355 strain was clustered together with Fusarium avenaceum, supported by 100%. According to the combined morphological characteristics and phylogenetic tree, the pathogen was identified as F. avenaceum. The mycelia of HGUP17355 strain grew fastest on oatmeal agar medium with maltose and L-phenylalanine as the carbon and nitrogen sources, respectively, at 20℃, pH 6.0, and its fatal temperature was 45℃ and it was insensitive to light.
To clarify the pathogen causing leaf brown spots of Chinese ground orchid Bletilla striata in Guizhou Province and its biological characteristics, the pathogen was obtained by using the method of tissue isolation. Its pathogenicity was tested in vitro inoculated leaves, and the pathogen was classified and identified with morphology and phylogeny based on combined ITS, EF-1α and β-tubulin sequence analysis; the biological characteristics of pathogen were also studied. The results showed that the strain isolated from leaf brown spots of B. striata was named as HGUP17355. Its main diagnostic features were as followed: white mycelia, cashmere-like form, uplifted center of colonies, faint yellow, maple-leaf red dorsal surface, oval microconidia with one-two septa, narrow sickle-like macroconidia with three-eight septa, and no chlamydospores. The phylogenetic tree based on combined ITS, EF-1α, β-tubulin sequences showed that HGUP17355 strain was clustered together with Fusarium avenaceum, supported by 100%. According to the combined morphological characteristics and phylogenetic tree, the pathogen was identified as F. avenaceum. The mycelia of HGUP17355 strain grew fastest on oatmeal agar medium with maltose and L-phenylalanine as the carbon and nitrogen sources, respectively, at 20℃, pH 6.0, and its fatal temperature was 45℃ and it was insensitive to light.
引文
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Kang X,Jin PF,Feng X,Liu WB,Zheng FC,Miao WG.2017.Identification and biological characteristics of the pathogen causing shoot blight of Moringa oleifera.Journal of Plant Protection,44(3):481-487(in Chinese)[康迅,靳鹏飞,冯霞,刘文波,郑服丛,缪卫国.2017.辣木枝枯病病原菌鉴定及其生物学特性.植物保护学报,44(3):481-487]
Kong Q,Yang SY,Wang CM,Xue CL,Guo JW,Li X,Zhou YL,Wu GM.2018.Pathogen identification and biological characteristics of black spot from Dendrobium officinale.Southwest China Journal of Agricultural Sciences,31(3):500-506(in Chinese)[孔琼,袁盛勇,王传铭,薛春丽,郭建伟,李珣,周银丽,吴光梅.2018.铁皮石斛黑斑病病原鉴定及生物学特性研究.西南农业学报,31(3):500-506]
Mackay GJ,Brown AE,Bjourson AJ,Mercer PC.1999.Molecular characterisation of Alternaria linicola and its detection in linseed.European Journal of Plant Pathology,105(2):157-166
O’Donnell K,Kistler HC,Cigelnik E,Ploetz RC.1998.Multiple evolutionary origins of the fungus causing Panama disease of banana:concordant evidence from nuclear and mitochondrial gene genealogies.Proceedings of the National Academy of Sciences of the United States of America,95(5):2044-2049
Parkunan V,Ji P.2013.First report of leaf spot on onion caused by Fusarium acuminatum in Georgia.Plant Disease,97(10):1384
Peever TL,Su G,Carpenter-Boggs L,Timmer LW.2004.Molecular systematics of citrus-associated Alternaria species.Mycologia,96(1):119-134
Ramdial HA,Rampersad SN.2010.First report of Fusarium solani causing fruit rot of sweet pepper in Trinidad.Plant Disease,94(11):1375
Ren J,Zeng GP,Song LS,Zhao Z,Li Z.2018.Pathogen identification and biological characteristics of Polygonum capitatum stem wilt.Acta Phytopathologica Sinica,48(2):271-274(in Chinese)[任静,曾桂萍,宋莉莎,赵致,李忠.2018.头花蓼茎枯病病原的鉴定及生物学特性.植物病理学报,48(2):271-274]
Ren J,Zeng GP,Zhao Z,Li Z.2016.A new species of Pilidiella on the Polygonaceae.Mycosystema,35(9):1056-1060(in Chinese)[任静,曾桂萍,赵致,李忠.2016.生于蓼科植物上的小帽壳孢属一新种.菌物学报,35(9):1056-1060]
Shi LJ.2014.The research of interaction mechanism of the main pathogenic fungi of dry rot in Heilongjiang Province.Master Thesis.Harbin:Northeast Agricultural University(in Chinese)[史丽娟.2014.黑龙江省马铃薯干腐病主要致病菌互作机制的研究.硕士学位论文.哈尔滨:东北农业大学]
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Stefańczyk E,Sobkowiak S,Brylińska M,?liwka J.2016.Diversity of Fusarium spp.associated with dry rot of potato tubers in Poland.European Journal of Plant Pathology,145(4):871-884
Su D.2013.The etiology and pathogenic mechanisms of medicinal plants windflower leaf spot.Master Thesis.Shenyang:Shenyang Agricultural University(in Chinese)[苏丹.2013.新归圃药用植物白头翁叶斑病病原学及其致病机理研究.硕士学位论文.沈阳:沈阳农业大学]
Su HR,He H,Huang QZ,Lu NH.2014.First report of black spot of Acanthus ilicifolius caused by Fusarium solani in China.Plant Disease,98(10):1438
Sun LL,Yang YH,Liu JK,Zhou M.2013.Bletilla striata pseudobulb rot pathogen identification and its biological characteristic.Journal of Chinese Medicinal Materials,36(3):341-345(in Chinese)[孙乐乐,杨永红,刘军凯,周敏.2013.白及根腐病病原鉴定和生物学特性研究.中药材,36(3):341-345]
Swofford DL.2002.PAUP*:phylogenetic analysis using parsimony(*and other methods).Sunderland,Massachusetts:Sinauer Associates,pp.54-56
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