吉林省大豆茎点霉叶斑病病原鉴定
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摘要
从吉林省白山市靖宇县的大豆叶片上分离到一种真菌分离物,根据菌落形态特征、分生孢子形态特征、ITS、ACT、tub2、rpb2、TEF、LSU序列、致病性测试结果以及上述6种引物的系统发育树构建分析,将其鉴定为Boeremia exigua var.exigua,这是在我国首次发现该真菌感染大豆。系统发育树分析结果中,rpb2和TEF基因在种间关系上显示出了较好的区分度,更适用于Boeremia属的鉴定及研究种间关系。
The strain was identified as Boeremia exigua var.exigua, which was collected form Jingyu County of Baishan City in Jilin, based on the morphological characteristics of the colony and conidia,rDNA-ITS,partial actin gene,beta-tubulin gene,RNA polymerase II second largest subunit gene,translation elongation factor 1 alpha gene,large subunit ribosomal RNA gene, the pathogenicity to young soybean, phylogenetic tree construction and analysis of the above 6 primers. It was the first report that Boeremia exigua var.exigua infected soybean in China.In the phylogenetic tree analysis results, rpb2 and TEF gene show a good discrimination in interspecies relationship, they are more suitable for identifying and studying intergenetic relationships of Boeremia.
The strain was identified as Boeremia exigua var.exigua, which was collected form Jingyu County of Baishan City in Jilin, based on the morphological characteristics of the colony and conidia,rDNA-ITS,partial actin gene,beta-tubulin gene,RNA polymerase II second largest subunit gene,translation elongation factor 1 alpha gene,large subunit ribosomal RNA gene, the pathogenicity to young soybean, phylogenetic tree construction and analysis of the above 6 primers. It was the first report that Boeremia exigua var.exigua infected soybean in China.In the phylogenetic tree analysis results, rpb2 and TEF gene show a good discrimination in interspecies relationship, they are more suitable for identifying and studying intergenetic relationships of Boeremia.
引文
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[4] 李绍兰,周斌,杨丽源,等.真菌DNA提取方法的改良[J].云南大学学报:自然科学版,2002,24 (6):471-472.(Li S L,Zhou B,Yang L Y,et al.An improved method for extracting fungal DNA[J].Journal of Yunnan University:Natural Sciences Edition,2002,24 (6):471-472.)
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[9] Groenewald J Z,Nakashima C,Nishikawa J,et al.Species concepts in Cercospora:Spotting the weeds among the roses[J].Studies in Mycology,2013,75(1):115-170.
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[11] Carbone I,Kohn L M.A method for designing primer sets for speciation studies in Filamentous Ascomycetes[J].Mycologia,1999,91(3):553-556.
[12] Aveskamp M M,de Gruyter J,Woudenberg J H,et al.Highlights of the Didymellaceae:A polyphasic approach to characterise Phoma and related pleosporalean genera[J].Studies in Mycology,2010,65(65):1-60.
[13] Schoch C L,Crous P W,Groenewald J Z,et al.A class-wide phylogenetic assessment of Dothideomycetes[J].Studies in Mycology,2009,64(64):1-15.
[14] Michel V V,Daepp M,Woudenberg J H C,et al.First report of Boeremia exigua var.exigua causing stem and leaf spot on common speedwell in Switzerland [J].Plant Disease,2018,102(2):440.
[15] Aveskamp M M,Verkley G J M,Gruyter J D,et al.DNA phylogeny reveals polyphyly of Phoma Section Peyronellaea and multiple Taxonomic Novelties[J].Mycologia,2009,101(3):363-382.
[16] de G J,Aveskamp M M,Woudenberg J H,et al.Molecular phylogeny of Phoma and allied anamorph genera:Towards a reclassification of the Phoma complex[J].Mycological Research,2009,113(4):508-519.
[17] Vu D,Groenewald M,Vries M D,et al.Large-scale generation and analysis of filamentous fungal DNA barcodes boosts coverage for kingdom Fungi and reveals thresholds for fungal species and higher taxon delimitation[J].Studies in Mycology,2019,92:135-154.
[18] Aveskamp M M,Woudenberg J H C,Gruyter J D,et al.Development of taxon-specific sequence characterized amplified region (SCAR) markers based on actin sequences and DNA amplification fingerprinting (DAF):A case study in the Phomaexigua species complex[J].Molecular Plant Pathology,2010,10(3):403-414.
[19] Kirk P,Cannon P,Minter D,et al.Dictionary of the fungi[J].Commonwealth Mycological Institute,2008:17-19.
[20] Boerema G H,Degruyter J,Noordeloos M E.Contributions towards a monograph of Phoma (Coelomycetes)—IV.Section Heterospora:Taxa with large sized conidial dimorphs,in vivo sometimes as Stagonosporopsis synanamorphs[J].Persoonia - Molecular Phylogeny and Evolution of Fungi,1997,16 (3):335-371.
[21] Chen Q,Jiang J R,Zhang G Z,et al.Resolving the Phoma enigma[J].Studies in Mycology,2015,82:137-217.
[22] Marcinkowska J,Roze-Katuny I,Katuny W.Pathogenicity of some Phomaexigua var.exigua isolates[J].Phytopathologia Polonica,2005(38):35-44.
[23] Zhao Q,Xie X W,Shi Y X,et al.Boeremia leaf and fruit spot of okra caused by Boeremia exigua in China[J].Canadian Journal of Plant Pathology,2016,38(3):395-399.
[24] Gai Y P,Ma H J,Chen X L,et al.Boeremia tuber rot of sweet potato caused by Boeremia exigua,a new post-harvest storage disease in China[J].Canadian Journal of Plant Pathology,2016,38(2):243-249.
[25] Thomidis T,Zambounis A,Prodromou I.Occurrence of a postharvest fruit rot disease of apples caused by Boeremia exigua var.exigua in Ptolemaida Kozani,Greece[J].Plant Disease,2016,100(11):2333.
[26] Samouel S,Iacovides T,Evangelides S,et al.First report of Boeremia exigua var.exigua causing stem rot of Origanum dubium in Cyprus[J].Plant Disease,2015,100(2):529.
[27] Gorny A M,Kikkert J R,Dunn A R,et al.Tan spot of lima bean caused by Boeremia exigua var.exigua in New York State,USA[J].Canadian Journal of Plant Pathology,2015,37(4):523-528.
[28] Li Y P,You M P,Finnegan P M,et al.First report of black spot caused by Boeremia exigua var.exigua on field pea in Australia[J].Plant Disease,2012,96(1):148-148.
[29] Jones S J,Hay F S,Harrington T C,et al.First report of boeremia blight caused by Boeremia exigua var.exigua on pyrethrum in Australia[J].Plant Disease,2011,95(11):1478-1478.
[30] 梁力哲.多变茎点菌的一个新变种[J].微生物学报,1991(2):79-81.(Liang L Z.A new variety of Phomaexigua[J].Actamicro Biologica Sinica,1991(2):79-81.)
[31] Mengistu A ,Sinclair J B .Seedborne micro-organisms of Ethiopian-grown soybean and chickpea seeds[J].Plant Disease Reporter,1979,63(7):616-619.
[32] Mendes M A S,daSilva V L,Dianese J C,et al.Fungos em plantas no Brasil[J].Brasillia:Embrapa,1998:555.
[33] Dingley J M,Fullerton R A,Mckenzie E H C.Survey of agricultural pests and diseases.Records of fungi,bacteria,algae and angiosperms pathogenic on plants in Cook Islands,Fiji,Kuribati,Niue,Tonga,Tuvalu and Western Samoa[J].Technical Report,1981,2.
[34] Rajak R,Rai M.Species of Phoma from legumes [J].Indian Phytopathol,1982,35:609-612.
[35] Alaka P,Rao V G.Compendium of fungi on legumes from India[J].Scientific Publishers (India),1998.
[36] Mutenko W,Majewski T,Ruszkiewicz-Michalska M,et al.A preliminary checklist of micromycetes in Poland[M]// Institute of Botany,Polish Academy of Sciences,2008,9:752.
[2] 马淑梅,丁俊杰,顾鑫,等.黑龙江省大豆主要病害发生危害调査[J].黑龙江农业科学,2005(6):48-52.(Ma S M,Ding J J,Gu X,et al.Survey on the occurrence and damage in main disease of soybean in Heilongjiang province[J].Heilongjiang Agricultural Science,2005(6):48-52.)
[3] 李沐慧,王媛媛,陈井生,等.2015年东北地区大豆田病害种类与危害程度调查研究[J].大豆科学,2016,35(4):643-648.(Li M H,Wang Y Y,Chen J S,et al.Incidence and disease index of soybean diseases in the Northeast of China in 2015[J].Soybean Science,2016,35(4):643-648.)
[4] 李绍兰,周斌,杨丽源,等.真菌DNA提取方法的改良[J].云南大学学报:自然科学版,2002,24 (6):471-472.(Li S L,Zhou B,Yang L Y,et al.An improved method for extracting fungal DNA[J].Journal of Yunnan University:Natural Sciences Edition,2002,24 (6):471-472.)
[5] White T J,Bruns T,Lee S,et al.Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics[M]// Innis M A,Gelfand D H,Sninsky J J,et al.Pcr Protocols:A guide to methods and applications.California:Academic Press,1990:315-322.
[6] Carbone I,Kohn L M.A method for designing primer sets for speciation studies in filamentous ascomycetes[J].Mycologia,1999:553-556.
[7] Vilgalys R,Hester M.Rapid genetic identification and mapping of enzymatically amplified ribosomal DNA from several Cryptococcus species[J].Journal of Bacteriology,1990,172(8):4238-4246.
[8] Rehner S A,Samuels G J.Taxonomy and phylogeny of Gliocladium,analysed from nuclear large subunit ribosomal DNA sequences[J].Mycological Research,1994,98(6):625-634.
[9] Groenewald J Z,Nakashima C,Nishikawa J,et al.Species concepts in Cercospora:Spotting the weeds among the roses[J].Studies in Mycology,2013,75(1):115-170.
[10] Liu Y J,Whelen S,Hall B D.Phylogenetic relationships among ascomycetes:Evidence from an RNA polymerse II subunit[J].Molecular Biology & Evolution,1999,16(12):1799.
[11] Carbone I,Kohn L M.A method for designing primer sets for speciation studies in Filamentous Ascomycetes[J].Mycologia,1999,91(3):553-556.
[12] Aveskamp M M,de Gruyter J,Woudenberg J H,et al.Highlights of the Didymellaceae:A polyphasic approach to characterise Phoma and related pleosporalean genera[J].Studies in Mycology,2010,65(65):1-60.
[13] Schoch C L,Crous P W,Groenewald J Z,et al.A class-wide phylogenetic assessment of Dothideomycetes[J].Studies in Mycology,2009,64(64):1-15.
[14] Michel V V,Daepp M,Woudenberg J H C,et al.First report of Boeremia exigua var.exigua causing stem and leaf spot on common speedwell in Switzerland [J].Plant Disease,2018,102(2):440.
[15] Aveskamp M M,Verkley G J M,Gruyter J D,et al.DNA phylogeny reveals polyphyly of Phoma Section Peyronellaea and multiple Taxonomic Novelties[J].Mycologia,2009,101(3):363-382.
[16] de G J,Aveskamp M M,Woudenberg J H,et al.Molecular phylogeny of Phoma and allied anamorph genera:Towards a reclassification of the Phoma complex[J].Mycological Research,2009,113(4):508-519.
[17] Vu D,Groenewald M,Vries M D,et al.Large-scale generation and analysis of filamentous fungal DNA barcodes boosts coverage for kingdom Fungi and reveals thresholds for fungal species and higher taxon delimitation[J].Studies in Mycology,2019,92:135-154.
[18] Aveskamp M M,Woudenberg J H C,Gruyter J D,et al.Development of taxon-specific sequence characterized amplified region (SCAR) markers based on actin sequences and DNA amplification fingerprinting (DAF):A case study in the Phomaexigua species complex[J].Molecular Plant Pathology,2010,10(3):403-414.
[19] Kirk P,Cannon P,Minter D,et al.Dictionary of the fungi[J].Commonwealth Mycological Institute,2008:17-19.
[20] Boerema G H,Degruyter J,Noordeloos M E.Contributions towards a monograph of Phoma (Coelomycetes)—IV.Section Heterospora:Taxa with large sized conidial dimorphs,in vivo sometimes as Stagonosporopsis synanamorphs[J].Persoonia - Molecular Phylogeny and Evolution of Fungi,1997,16 (3):335-371.
[21] Chen Q,Jiang J R,Zhang G Z,et al.Resolving the Phoma enigma[J].Studies in Mycology,2015,82:137-217.
[22] Marcinkowska J,Roze-Katuny I,Katuny W.Pathogenicity of some Phomaexigua var.exigua isolates[J].Phytopathologia Polonica,2005(38):35-44.
[23] Zhao Q,Xie X W,Shi Y X,et al.Boeremia leaf and fruit spot of okra caused by Boeremia exigua in China[J].Canadian Journal of Plant Pathology,2016,38(3):395-399.
[24] Gai Y P,Ma H J,Chen X L,et al.Boeremia tuber rot of sweet potato caused by Boeremia exigua,a new post-harvest storage disease in China[J].Canadian Journal of Plant Pathology,2016,38(2):243-249.
[25] Thomidis T,Zambounis A,Prodromou I.Occurrence of a postharvest fruit rot disease of apples caused by Boeremia exigua var.exigua in Ptolemaida Kozani,Greece[J].Plant Disease,2016,100(11):2333.
[26] Samouel S,Iacovides T,Evangelides S,et al.First report of Boeremia exigua var.exigua causing stem rot of Origanum dubium in Cyprus[J].Plant Disease,2015,100(2):529.
[27] Gorny A M,Kikkert J R,Dunn A R,et al.Tan spot of lima bean caused by Boeremia exigua var.exigua in New York State,USA[J].Canadian Journal of Plant Pathology,2015,37(4):523-528.
[28] Li Y P,You M P,Finnegan P M,et al.First report of black spot caused by Boeremia exigua var.exigua on field pea in Australia[J].Plant Disease,2012,96(1):148-148.
[29] Jones S J,Hay F S,Harrington T C,et al.First report of boeremia blight caused by Boeremia exigua var.exigua on pyrethrum in Australia[J].Plant Disease,2011,95(11):1478-1478.
[30] 梁力哲.多变茎点菌的一个新变种[J].微生物学报,1991(2):79-81.(Liang L Z.A new variety of Phomaexigua[J].Actamicro Biologica Sinica,1991(2):79-81.)
[31] Mengistu A ,Sinclair J B .Seedborne micro-organisms of Ethiopian-grown soybean and chickpea seeds[J].Plant Disease Reporter,1979,63(7):616-619.
[32] Mendes M A S,daSilva V L,Dianese J C,et al.Fungos em plantas no Brasil[J].Brasillia:Embrapa,1998:555.
[33] Dingley J M,Fullerton R A,Mckenzie E H C.Survey of agricultural pests and diseases.Records of fungi,bacteria,algae and angiosperms pathogenic on plants in Cook Islands,Fiji,Kuribati,Niue,Tonga,Tuvalu and Western Samoa[J].Technical Report,1981,2.
[34] Rajak R,Rai M.Species of Phoma from legumes [J].Indian Phytopathol,1982,35:609-612.
[35] Alaka P,Rao V G.Compendium of fungi on legumes from India[J].Scientific Publishers (India),1998.
[36] Mutenko W,Majewski T,Ruszkiewicz-Michalska M,et al.A preliminary checklist of micromycetes in Poland[M]// Institute of Botany,Polish Academy of Sciences,2008,9:752.