北京水系中腐霉和疫腐霉种类调查
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摘要
为研究北京地区水系中腐霉和疫腐霉种类分布,采用叶饵法从北京水系中诱集菌株,共分离到腐霉、疫腐霉182株,利用Single-strand conformation polymorphism (SSCP)进行分析,筛选出不同基因型的代表菌株,并依据形态学特征及ITS序列对这些菌株进行种类鉴定。结果共发现6个已知种和3个可能的新种,其中:湖滨疫腐霉Phytopythium litorale和果胶裂解腐霉Pythium pectinolyticum为国内新记录种;旋柄疫腐霉Pp.helicoides、异丝腐霉P.diclinum、簇囊腐霉P.torulosum和袋囊腐霉P.marsipium为北京地区新记录种。研究结果可为未来监测北京地区水系中病原菌的分布情况提供基础数据。
To investigate the species composition of Pythium and Phytopythium in Beijing water system,strains of Pythium and Phytopythium were collected from the Beijing river system by leaf baiting method.A total of 182 strains were isolated and analyzed by Single-strand conformation polymorphism(SSCP).Representative strains of different genotypes were then selected and identified base on their morphological characteristics and ITS sequences.The resulted showed that six known species and three suspected new species were identified.Among them,Phytopythium litotale and Pythium pectinolyticum were new records in China,while Pp.helicoides,P.diclinum,P.torulosum and P.marsipium were new records in Beijing.This study provided basic data for the future monitoring of the plant pathogen in Beijing water system.
To investigate the species composition of Pythium and Phytopythium in Beijing water system,strains of Pythium and Phytopythium were collected from the Beijing river system by leaf baiting method.A total of 182 strains were isolated and analyzed by Single-strand conformation polymorphism(SSCP).Representative strains of different genotypes were then selected and identified base on their morphological characteristics and ITS sequences.The resulted showed that six known species and three suspected new species were identified.Among them,Phytopythium litotale and Pythium pectinolyticum were new records in China,while Pp.helicoides,P.diclinum,P.torulosum and P.marsipium were new records in Beijing.This study provided basic data for the future monitoring of the plant pathogen in Beijing water system.
引文
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[2]Kirk P M,Cannon P F,Minter D W,Stalpers J A.Dictionary of the Fungi[M].10th ed.Walling ford:Center for Applied Biological International,2008
[3]Sanogo S,Moorman G W.Transmission and control of Pythium aphanidermatum in an ebb-and-flow subirrigation system[J].Plant Disease,1993,77(3):287-290
[4]Cock A W,Lodhi A M,Rintoul T L,Bala K,Robideau G P,Gloria A Z,Coffey M D,Shahzad S,Lévesque C A.Phytopythium:Molecular phylogeny and systematics[J].Persoonia Molecular Phylogeny and Evolution of Fungi,2015,34:25-39
[5]Ho H H.The genus Pythiumin Taiwan,China:A synoptic review[J].Frontiers of Biology in China,2009,4(1):15-28
[6]Ho H H.The genus Pythium in mainland China[J].Mycosystema,2013,32(1):21-44
[7]Bush E A,Hong C X,Stromberg E L.Fluctuations of Phytophthoraand Pythiumspp in components of a recycling irrigation system[J].Plant Disease,2003,87(12):1500-1506
[8]Dick M W.Straminopilous fungi:Systematics of the Peronosporomycetes including accounts of the marine of novel filamentous morphology[J].Journal of General Virology,2001,75:3585-3590
[9]Ho H H.The taxonomy and biology of Phytophthora and Pythium[J].Journal of Bacteriology and Mycology,2018,6(2):1-7
[10]Middleton J T.The taxonomy,host range and geographic distribution of the genus Pythium[J].Memoirs of the Torrey Botanical Club,1943,20(1):1-171
[11]Waterhouse G E.Key to Pythiumpringsheim:Commonwealth mycological institute[J].Mycological Papers,1967,109:1-15
[12]vander Plaats-Niterink A J.Monograph of the genus Pythium[J].Studies in Mycology,1981,5(21):1-242
[13]Dick M W.Keys to Pythium[M].Reading:University of Reading,1990
[14]Kroon L P N M,Bakker F T,Bosch G B M,Bonants P J M,Flier W G.Phylogenetic analysis of Phytophthora species based on mitochondrial and nuclear DNA sequences[J].Fungal Genetics and Biology,2004,41(8):766-782
[15]Lévesque C A,Cock A W.Molecular phylogeny and taxonomy of the genus Pythium[J].Mycological Research,2004,108(12):1363-1383
[16]Kong P,Richardson P A,Moorman G W,Hong C X.Singlestrand conformational polymorphism analysis of the ribosomal internal transcribed spacer 1for rapid species identification within the genus Pythium[J].FEMS Microbiology Letters,2004,240(2):229-236
[17]Ghimire S R,Richardson P A,Moorman G W,Lea-Cox J D,Ross D S,Hong C X.An in-situ baiting bioassay for detecting Phytophthora species in irrigation runoff containment basins[J].Plant Pathology,2009,58(3):577-583
[18]郑小波．疫霉菌及其研究技术[M]．北京：中国农业出版社，2000Zheng X B.Phytophthora and Its Research Technique[M].Beijing:China Agriculture Press,1997(in Chinese）
[19]Guo L Y,Ko W H.Two widely accessible media for growth and reproduction of Phytophthora and Pythium species[J].Applied Environmental Microbiology,1993,59(7):2323-2325
[20]Kong P,Richardson P A,Hong C X.Direct colony PCR-SSCPfor detection of multiplepythiaceous oomycetes in environmental samples[J].Journal of Microbiological Methods,2005,61(1):25-32
[21]Wang Q M,Li J,Wang S A,Bai F Y.Rapid differentiation of phenotypically similar yeast species by single-strand conformation polymorphism of ribosomal DNA[J].Applied and Environmental Microbiology,2008,74(9):2604-2611
[22]Ribeiro O K.A Source Book of the Genus Phytophthora[M].Washington:Quarterly Review of Biology,1978．
[23]Guo L Y,Zhu X Q,Hu C H,Ristaino J B.Genetic structure of Phytophthora infestans populations in China indicates multiple migration events[J].Phytopathology,2010,100(10):997-1006
[24]Cooke D E L,Duncan J M.Phylogenetic analysis of Phytophthoraspecies based on ITS1and ITS2sequences of the ribosomal RNA gene repeat[J].Mycological Research,1997,101(6):667-677
[25]曹欣然，朱小琼，国立耘．分离自北京水系的疫霉属三个中国新记录种[J]．菌物学报，2017,36(12):1697-1707Cao X R,Zhu X Q,Guo L Y.Three Chinese new records of Phytophthora isolated from Beijing water system[J].Mycosystema,2017,36(12):1697-1707(in Chinese）
[26]Thompson J D,Gibson T J,Plewniak F,Jeanmougin F,Higgins D G.The Clustal X windows interface:Flexible strategies for multiple sequence alignment aided by quality analysis tools[J].Nucleic Acids Research,1997,25(24):4876-4882
[27]Kumar,S,Tamura K,Nei M.MEGA3:Integrated software for molecular evolutionary genetics analysis and sequence alignment[J].Briefings in Bioinformatics,2004,5(2):150-163
[28]Nechwatal J,Mendgen K.Pythium litorale sp nov,a new species from the littoral of lake constance Germany[J].FEMSMicrobiology Letters,2006,255(1):96-101
[29]Chenari Bouket A,Babai-Ahari A,Arzanlou M,Tojo M.Morphological and molecular characterization of Phytopythium litorale and Pp oedochilum from Iran[J].Nova Hedwigia,2016,102(1-2):257-270
[30]Paul B.A new species of Pythium with filamentous sporangia having pectinolytic activities,isolated in the Burgundy region of France[J].FEMS Microbiology Letters,2001,199(1):55-59
[31]Uzuhashi S,Okada G,Ohkuma M.Four new Pythiumspecies from aquatic environments in Japan[J].Antonie van Leeuwenhoek,2015,107(2):375-391
[32]Ho HH,Chen X X,Zeng H C,Zheng F C.The occurrence and distribution of Pythium species on Hainan island of south China[J].Botanical Studies,2012,53(4):525-534
[33]余永年．中国真菌志第六卷霜霉目[M]．北京：科学出版社，1998Yu Y N.Sinomycota:The Sixth Volume Peronosporales[M].Beijing:Science Press,1998(in Chinese）
[34]Yu Y N,Ma G Z.The genus Pythium in China[J].Mycosystema,1989,2:1-110
[35]Al-Sheikh H.Two pathogenic species of Pythium:Paphanidermatum and P diclinum from a wheat field[J].Saudi Journal of Biological Sciences,2010,17(4):347-352
[36]龙艳艳．腐霉属的DNA条形码和分子系统学研究[D]．南宁：广西大学，2014Long Y Y.DNA Barcoding and molecular systematic of Pythium[D].Nanning:Guangxi University,2014(in Chinese）
[37]Ho H H.The genus Pythiumin Taiwan(2):An illustrated diagnostic key[J].Mycotaxon,2011,116(1):33-47
[2]Kirk P M,Cannon P F,Minter D W,Stalpers J A.Dictionary of the Fungi[M].10th ed.Walling ford:Center for Applied Biological International,2008
[3]Sanogo S,Moorman G W.Transmission and control of Pythium aphanidermatum in an ebb-and-flow subirrigation system[J].Plant Disease,1993,77(3):287-290
[4]Cock A W,Lodhi A M,Rintoul T L,Bala K,Robideau G P,Gloria A Z,Coffey M D,Shahzad S,Lévesque C A.Phytopythium:Molecular phylogeny and systematics[J].Persoonia Molecular Phylogeny and Evolution of Fungi,2015,34:25-39
[5]Ho H H.The genus Pythiumin Taiwan,China:A synoptic review[J].Frontiers of Biology in China,2009,4(1):15-28
[6]Ho H H.The genus Pythium in mainland China[J].Mycosystema,2013,32(1):21-44
[7]Bush E A,Hong C X,Stromberg E L.Fluctuations of Phytophthoraand Pythiumspp in components of a recycling irrigation system[J].Plant Disease,2003,87(12):1500-1506
[8]Dick M W.Straminopilous fungi:Systematics of the Peronosporomycetes including accounts of the marine of novel filamentous morphology[J].Journal of General Virology,2001,75:3585-3590
[9]Ho H H.The taxonomy and biology of Phytophthora and Pythium[J].Journal of Bacteriology and Mycology,2018,6(2):1-7
[10]Middleton J T.The taxonomy,host range and geographic distribution of the genus Pythium[J].Memoirs of the Torrey Botanical Club,1943,20(1):1-171
[11]Waterhouse G E.Key to Pythiumpringsheim:Commonwealth mycological institute[J].Mycological Papers,1967,109:1-15
[12]vander Plaats-Niterink A J.Monograph of the genus Pythium[J].Studies in Mycology,1981,5(21):1-242
[13]Dick M W.Keys to Pythium[M].Reading:University of Reading,1990
[14]Kroon L P N M,Bakker F T,Bosch G B M,Bonants P J M,Flier W G.Phylogenetic analysis of Phytophthora species based on mitochondrial and nuclear DNA sequences[J].Fungal Genetics and Biology,2004,41(8):766-782
[15]Lévesque C A,Cock A W.Molecular phylogeny and taxonomy of the genus Pythium[J].Mycological Research,2004,108(12):1363-1383
[16]Kong P,Richardson P A,Moorman G W,Hong C X.Singlestrand conformational polymorphism analysis of the ribosomal internal transcribed spacer 1for rapid species identification within the genus Pythium[J].FEMS Microbiology Letters,2004,240(2):229-236
[17]Ghimire S R,Richardson P A,Moorman G W,Lea-Cox J D,Ross D S,Hong C X.An in-situ baiting bioassay for detecting Phytophthora species in irrigation runoff containment basins[J].Plant Pathology,2009,58(3):577-583
[18]郑小波．疫霉菌及其研究技术[M]．北京：中国农业出版社，2000Zheng X B.Phytophthora and Its Research Technique[M].Beijing:China Agriculture Press,1997(in Chinese）
[19]Guo L Y,Ko W H.Two widely accessible media for growth and reproduction of Phytophthora and Pythium species[J].Applied Environmental Microbiology,1993,59(7):2323-2325
[20]Kong P,Richardson P A,Hong C X.Direct colony PCR-SSCPfor detection of multiplepythiaceous oomycetes in environmental samples[J].Journal of Microbiological Methods,2005,61(1):25-32
[21]Wang Q M,Li J,Wang S A,Bai F Y.Rapid differentiation of phenotypically similar yeast species by single-strand conformation polymorphism of ribosomal DNA[J].Applied and Environmental Microbiology,2008,74(9):2604-2611
[22]Ribeiro O K.A Source Book of the Genus Phytophthora[M].Washington:Quarterly Review of Biology,1978．
[23]Guo L Y,Zhu X Q,Hu C H,Ristaino J B.Genetic structure of Phytophthora infestans populations in China indicates multiple migration events[J].Phytopathology,2010,100(10):997-1006
[24]Cooke D E L,Duncan J M.Phylogenetic analysis of Phytophthoraspecies based on ITS1and ITS2sequences of the ribosomal RNA gene repeat[J].Mycological Research,1997,101(6):667-677
[25]曹欣然，朱小琼，国立耘．分离自北京水系的疫霉属三个中国新记录种[J]．菌物学报，2017,36(12):1697-1707Cao X R,Zhu X Q,Guo L Y.Three Chinese new records of Phytophthora isolated from Beijing water system[J].Mycosystema,2017,36(12):1697-1707(in Chinese）
[26]Thompson J D,Gibson T J,Plewniak F,Jeanmougin F,Higgins D G.The Clustal X windows interface:Flexible strategies for multiple sequence alignment aided by quality analysis tools[J].Nucleic Acids Research,1997,25(24):4876-4882
[27]Kumar,S,Tamura K,Nei M.MEGA3:Integrated software for molecular evolutionary genetics analysis and sequence alignment[J].Briefings in Bioinformatics,2004,5(2):150-163
[28]Nechwatal J,Mendgen K.Pythium litorale sp nov,a new species from the littoral of lake constance Germany[J].FEMSMicrobiology Letters,2006,255(1):96-101
[29]Chenari Bouket A,Babai-Ahari A,Arzanlou M,Tojo M.Morphological and molecular characterization of Phytopythium litorale and Pp oedochilum from Iran[J].Nova Hedwigia,2016,102(1-2):257-270
[30]Paul B.A new species of Pythium with filamentous sporangia having pectinolytic activities,isolated in the Burgundy region of France[J].FEMS Microbiology Letters,2001,199(1):55-59
[31]Uzuhashi S,Okada G,Ohkuma M.Four new Pythiumspecies from aquatic environments in Japan[J].Antonie van Leeuwenhoek,2015,107(2):375-391
[32]Ho HH,Chen X X,Zeng H C,Zheng F C.The occurrence and distribution of Pythium species on Hainan island of south China[J].Botanical Studies,2012,53(4):525-534
[33]余永年．中国真菌志第六卷霜霉目[M]．北京：科学出版社，1998Yu Y N.Sinomycota:The Sixth Volume Peronosporales[M].Beijing:Science Press,1998(in Chinese）
[34]Yu Y N,Ma G Z.The genus Pythium in China[J].Mycosystema,1989,2:1-110
[35]Al-Sheikh H.Two pathogenic species of Pythium:Paphanidermatum and P diclinum from a wheat field[J].Saudi Journal of Biological Sciences,2010,17(4):347-352
[36]龙艳艳．腐霉属的DNA条形码和分子系统学研究[D]．南宁：广西大学，2014Long Y Y.DNA Barcoding and molecular systematic of Pythium[D].Nanning:Guangxi University,2014(in Chinese）
[37]Ho H H.The genus Pythiumin Taiwan(2):An illustrated diagnostic key[J].Mycotaxon,2011,116(1):33-47
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