浙江雁荡山山脉叶栖酵母菌资源与物种多样性
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摘要
对北雁荡山、中雁荡山、南雁荡山、西雁荡山和东雁荡山等广阔区域内的叶栖酵母菌资源进行考察,旨在弄清雁荡山山脉叶栖酵母菌资源和多样性。采用PDA平板对雁荡山地区的叶栖酵母菌资源进行分离和纯化,根据酵母菌培养物26S rDNA D1/D2序列和系统发育关系明确酵母菌的分类地位,分析该地区叶栖酵母菌群落的多样性、组成和分布情况。结果发现,分离自雁荡山山脉的1118株叶栖酵母菌可以被鉴定为56个已知类群和19个疑似新种,其中38种酵母菌在该地区比较常见。常见的酵母菌类群中,包括Derxomyces属5种,Sporobolomyces属3种,Bannoa、Bulleribasidium、Rhodosporidiobolus、Symmetrospora、Taphrina、Tilletiopsi和Udeniomyces属各2种,Bullera、Coniochaeta、Coniosporium、Cryptococcus、Elsinoe、Erythrobasidium、Fellozyma、Golubevia、Kockovaella、Kondoa、Leucosporidium、Moesziomyces、Oberwinklerozyma、Phyllozyma、Ruinenia和Saitozyma属各1种。根据常见酵母菌类群的相对频率,在该地区分离到的叶栖酵母菌中Derxomyces mrakii具有明显分布优势,并探讨了Derxomyces mrakii株间26S rDNA D1/D2序列的稳定性和变异幅度。根据雁荡山地区常见叶栖酵母菌组成和相似程度,可以推测地理上相近地区的酵母菌物种分布具有更大相似性。基于雁荡山山脉常见叶栖酵母菌种类和数量而绘制出的物种累积曲线,证明雁荡山地区绝大多数的叶栖酵母菌种类已经被分离到,研究结果可以真实反映该地区叶栖酵母菌资源和物种多样性的存在状况。
Phyllosphere yeasts from the Yandang Mountains were investigated to reveal their resources and species diversity.The phyllosphere yeasts from the focal region were isolated and purified in a potato dextrose agar( PDA) plate and their classification statuses were defined according to their 26 S rDNA D1/D2 sequences and the phylogenetic relationships of thecultures,and then the composition,distribution,and diversity of the phyllosphere yeasts were analyzed in detail. In total,1118 phyllosphere yeasts were isolated from the Yandang Mountains and identified as 56 known groups and 19 proposed new species,among which 38 groups were common. The common yeast species were widely distributed among 25 genera. Of these, the Derxomyces consisted of five species, the Sporobolomyces consisted of three species, the Bannoa,Bulleribasidium,Rhodosporidiobolus, Symmetrospora, Taphrina, Tilletiopsis, and Udeniomyces each consisted of two species,whereas the Bullera,Coniochaeta,Coniosporium,Cryptococcus,Elsinoe,Erythrobasidium,Fellozyma,Golubevia,Kockovaella,Kondoa, Leucosporidium, Moesziomyces, Oberwinklerozyma, Phyllozyma, Ruinenia, and Saitozyma each consisted of one species. According to the relative frequency of the common groups,Derxomyces mrakii had an obvious distribution dominance. At the same time,the stability and variation range of the 26 S rDNA D1/D2 sequence among the different strains of D. mrakii were discussed. Based on a comparison of the composition and similarity of common phyllosphere yeasts in the Yandang Mountains, it can be speculated that the distribution of yeast species within geographically close regions might have more similarities. The majority of the phyllosphere yeasts in Yandang Mountains were isolated through analyzing the species accumulation curves based on the species and numbers of common phyllosphere yeasts. Therefore,this study reflects resources and species diversity of the phyllosphere yeasts from the Yandang Mountains.
Phyllosphere yeasts from the Yandang Mountains were investigated to reveal their resources and species diversity.The phyllosphere yeasts from the focal region were isolated and purified in a potato dextrose agar( PDA) plate and their classification statuses were defined according to their 26 S rDNA D1/D2 sequences and the phylogenetic relationships of thecultures,and then the composition,distribution,and diversity of the phyllosphere yeasts were analyzed in detail. In total,1118 phyllosphere yeasts were isolated from the Yandang Mountains and identified as 56 known groups and 19 proposed new species,among which 38 groups were common. The common yeast species were widely distributed among 25 genera. Of these, the Derxomyces consisted of five species, the Sporobolomyces consisted of three species, the Bannoa,Bulleribasidium,Rhodosporidiobolus, Symmetrospora, Taphrina, Tilletiopsis, and Udeniomyces each consisted of two species,whereas the Bullera,Coniochaeta,Coniosporium,Cryptococcus,Elsinoe,Erythrobasidium,Fellozyma,Golubevia,Kockovaella,Kondoa, Leucosporidium, Moesziomyces, Oberwinklerozyma, Phyllozyma, Ruinenia, and Saitozyma each consisted of one species. According to the relative frequency of the common groups,Derxomyces mrakii had an obvious distribution dominance. At the same time,the stability and variation range of the 26 S rDNA D1/D2 sequence among the different strains of D. mrakii were discussed. Based on a comparison of the composition and similarity of common phyllosphere yeasts in the Yandang Mountains, it can be speculated that the distribution of yeast species within geographically close regions might have more similarities. The majority of the phyllosphere yeasts in Yandang Mountains were isolated through analyzing the species accumulation curves based on the species and numbers of common phyllosphere yeasts. Therefore,this study reflects resources and species diversity of the phyllosphere yeasts from the Yandang Mountains.
引文
[1]Kurtzman C P,Fell J W,Boekhout T.Definition,classification and nomenclature of the yeasts//Kurtzman C P,Fell J W,Boekhout T,eds.The yeasts:A Taxonomic Study.5th ed.Amsterdam:Elsevier,2011:3-5.
[2]Alexopoulos C J,Mims C W,Blackwell M M.Introductory Mycology.4th ed.New York:John Wiley&Sons Inc.,1996:1-880.
[3]Bhadra B,Rao R S,Singh P K,Sarkar P K,Shivaji S.Yeasts and yeast-like fungi associated with tree bark:diversity and identification of yeasts producing extracellular endoxylanases.Current Microbiology,2008,56(5):489-494.
[4]Valdez A V,Garcia L S,Kirchmayr M,Rodríguez P R,Esquinca A G,Coria R,Mathis A G.Yeast communities associated with artisanal mezcal fermentations from Agave salmiana.Antonie van Leeuwenhoek,2011,100(4):497-506.
[5]Bezerra-Bussoli C,Baffi M A,Gomes E,Da-Silva R.Yeast diversity isolated from grape musts during spontaneous fermentation from a Brazilian winery.Current Microbiology,2013,67(3):356-361.
[6]?eni?ováK,ChovanováK,Chebeňová-TurcovskáV,GodálováZ,KrakováL,Kuchta T,Pangallo D,Bre6náB.Mapping of wine yeast and fungal diversity in the Small Carpathian wine-producing region(Slovakia):evaluation of phenotypic,genotypic and culture-independent approaches.Annals of Microbiology,2014,64(4):1819-1828.
[7]Liu R,Zhang Q H,Chen F S,Zhang X Y.Analysis of culturable yeast diversity in spontaneously fermented orange wine,orange peel and orangery soil of a Ponkan plantation in China.Annals of Microbiology,2015,65(4):2387-2391.
[8]Pathania N,Kanwar S S,Jhang T,Koundal K R,Sharma T R.Application of different molecular techniques for deciphering genetic diversity among yeast isolates of traditional fermented food products of Western Himalayas.World Journal of Microbiology and Biotechnology,2010,26(9):1539-1547.
[9]Oguntoyinbo F A.Culture-independent analysis for determination of yeast diversity during solid substrate fermentation of grated cassava for gari production.World Journal of Microbiology and Biotechnology,2011,27(10):2461-2465.
[10]Haque M A,Seo W T,Hwang C E,Lee H Y,Ahn M J,Cho K M.Culture-independent analysis of yeast diversity in Korean traditional fermented soybean foods(doenjang and kanjang)based on 26S rRNA sequence.Journal of the Korean Society for Applied Biological Chemistry,2015,58(3):377-385.
[11]Rosa C A,Resende M A,Barbosa F A R,Morais P B,Franzot S P.Yeast diversity in a mesotrophic lake on the karstic plateau of Lagoa Santa,MG-Brazil.Hydrobiologia,1995,308(2):103-108.
[12]Yurkov A M,Wehde T,Federici J,Sch?fer A M,Ebinghaus M,Lotze-Engelhard S,Mittelbach M,Prior R,Richter C,R9hl O,Begerow D.Yeast diversity and species recovery rates from beech forest soils.Mycological Progress,2016,15(8):845-859.
[13]Pozo M I,Herrera C M,Bazaga P.Species richness of yeast communities in floral nectar of southern Spanish plants.Microbial Ecology,2011,61(1):82-91.
[14]Kachalkin A V,Yurkov A M.Yeast communities in Sphagnum phyllosphere along the temperature-moisture ecocline in the boreal forest-swamp ecosystem and description of Candida sphagnicola sp.nov.Antonie van Leeuwenhoek,2012,102(1):29-43.
[15]Nasanit R,Jaibangyang S,Tantirungkij M,Limtong S.Yeast diversity and novel yeast D1/D2 sequences from corn phylloplane obtained by a culture-independent approach.Antonie van Leeuwenhoek,2016,109(12):1615-1634.
[16]Wang H M,Xu J P,Guo H,Wu J Y,Yi G H,Pei H,Niu L N,Li Y.Patterns of human oral yeast species distribution on Hainan Island in China.Mycopathologia,2013,176(5/6):359-368.
[17]Faria-Ramos I,Neves-Maia J,Ricardo E,Santos-Antunes J,Silva A T,Costa-de-Oliveira S,Cantón E,Rodrigues A G,Pina-Vaz C.Species distribution and in vitro antifungal susceptibility profiles of yeast isolates from invasive infections during a Portuguese multicenter survey.European Journal of Clinical Microbiology&Infectious Diseases,2014,33(12):2241-2247.
[18]Li W,Hu Y A,Li F Q,Shi L N,Shao H F,Huang M,Wang Y,Han D D,Liao H,Ma C F,Zhang G Y.Distribution of Yeast Isolates from Invasive Infections and Their in vitro susceptibility to antifungal agents:evidence from 299 cases in a 3-year(2010 to 2012)surveillance study.Mycopathologia,2015,179(5/6):397-405.
[19]Gadanho M,Libkind D,Sampaio J.Yeast diversity in the extreme acidic environments of the Iberian pyrite belt.Microbial Ecology,2006,52(3):552-563.
[20]Gümral R,Tümg?r A,Sara?l?M A,Y?ld?ranST,Ilkit M,de Hoog G S.Black yeast diversity on creosoted railway sleepers changes with ambient climatic conditions.Microbial Ecology,2014,68(4):699-707.
[21]Nakase T,Takashima M.A simple procedure for the high frequency isolation of new taxa of ballistosporous yeasts living on the surface of plants.RIKEN Review,1993,3:33-34.
[22]Makimura K,Murayama S Y,Yamaguchi H.Detection of a wide range of medically important fungi by the polymerase chain reaction.Journal of Medical Microbiology,1994,40(5):358-364.
[23]Kurtzman C P,Robnett C J.Identification of clinically important ascomycetous yeasts based on nucleotide divergence in the 5'end of the largesubunit(26S)ribosomal DNA gene.Journal of Clinical Microbiology,1997,35(5):1216-1223.
[24]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.Nucleic Acids Research,1997,25(24):4876-4882.
[25]Saitou N,Nei M.The neighbor-joining method:a new method for reconstructing phylogenetic trees.Molecular Biology and Evolution,1987,4(4):406-425.
[26]Page R D M.Tree View:an application to display phylogenetic trees on personal computers.Bioinformatics,1996,12(4):357-358.
[27]Kurtzman C P,Robnett C J.Identification and phylogeny of ascomycetous yeasts from analysis of nuclear large subunit(26S)ribosomal DNA partial sequences.Antonie van Leeuwenhoek,1998,73(4):331-371.
[28]Fell J W,Boekhout T,Fonseca A,Scorzetti G,Statzell-Tallman A.Biodiversity and systematics of basidiomycetous yeasts as determined by largesubunit r DNA D1/D2 domain sequence analysis.International Journal of Systematic and Evolutionary Microbiology,2000,50(3):1351-1371.
[29]Scorzetti G,Fell J W,Fonseca A,Statzell-Tallman A.Systematics of basidiomycetous yeasts:a comparison of large subunit D1/D2 and internal transcribed spacer r DNA regions.FEMS Yeast Research,2002,2(4):495-571.
[30]Dong L,Zhang J,Sun Y,Liu Y,Zhang Y Y,Zheng G M.Phylogeographic patterns and conservation units of a vulnerable species,Cabot's tragopan(Tragopan caboti),endemic to Southeast China.Conservation Genetics,2010,11(6):2231-2242.
[31]Glushakova A M,Chernov I Y.Seasonal dynamics of the structure of epiphytic yeast communities.Microbiology,2010,79(6):830-839.
[32]Han P J,Li A H,Wang Q M,Bai F Y.Ballistosporomyces changbaiensis sp.nov.and Ballistosporomyces bomiensis sp.nov.,two novel species isolated from shrub plant leaves.Antonie van Leeuwenhoek,2016,109(7):965-970.
[33]Glushakova A M,Chernov I Y.Seasonal dynamics in a yeast population on leaves of the common wood sorrel Oxalis acetosella L.Microbiology,2004,73(2):184-188.
[34]Liu X Z,Wang Q M,G9ker M,Groenewald M,Kachalkin A V,Lumbsch H T,Millanes A M,Wedin M,Yurkov A M,Boekhout T,Bai F Y.Towards an integrated phylogenetic classification of the Tremellomycetes.Studies in Mycology,2015,81:85-147.
[35]Wang Q M,Begerow D,Groenewald M,Liu X Z,Theelen B,Bai F Y,Boekhout T.Multigene phylogeny and taxonomic revision of yeasts and related fungi in the Ustilaginomycotina.Studies in Mycology,2015,81:55-83.
[36]Wang Q M,Yurkov A M,G9ker M,Lumbsch H T,Leavitt S D,Groenewald M,Theelen B,Liu X Z,Boekhout T,Bai F Y.Phylogenetic classification of yeasts and related taxa within Pucciniomycotina.Studies in Mycology,2015,81:149-189.
[37]Hamamoto M,Nakase T.Ballistosporous yeasts found on the surface of plant materials collected in New Zealand.Antonie van Leeuwenhoek,1996,69(3):279-291.
[38]Wang Q M,Bai F Y.Molecular phylogeny of basidiomycetous yeasts in the Cryptococcus luteolus lineage(Tremellales)based on nuclear rRNA and mitochondrial cytochrome b gene sequence analyses:proposal of Derxomyces gen.nov.and Hannaella gen.nov.,and description of eight novel Derxomyces species.FEMS Yeast Research,2008,8(5):799-814.
[39]Smith E P,van Belle G.Nonparametric estimation of species richness.Biometrics,1984,40(1):119-129.
[2]Alexopoulos C J,Mims C W,Blackwell M M.Introductory Mycology.4th ed.New York:John Wiley&Sons Inc.,1996:1-880.
[3]Bhadra B,Rao R S,Singh P K,Sarkar P K,Shivaji S.Yeasts and yeast-like fungi associated with tree bark:diversity and identification of yeasts producing extracellular endoxylanases.Current Microbiology,2008,56(5):489-494.
[4]Valdez A V,Garcia L S,Kirchmayr M,Rodríguez P R,Esquinca A G,Coria R,Mathis A G.Yeast communities associated with artisanal mezcal fermentations from Agave salmiana.Antonie van Leeuwenhoek,2011,100(4):497-506.
[5]Bezerra-Bussoli C,Baffi M A,Gomes E,Da-Silva R.Yeast diversity isolated from grape musts during spontaneous fermentation from a Brazilian winery.Current Microbiology,2013,67(3):356-361.
[6]?eni?ováK,ChovanováK,Chebeňová-TurcovskáV,GodálováZ,KrakováL,Kuchta T,Pangallo D,Bre6náB.Mapping of wine yeast and fungal diversity in the Small Carpathian wine-producing region(Slovakia):evaluation of phenotypic,genotypic and culture-independent approaches.Annals of Microbiology,2014,64(4):1819-1828.
[7]Liu R,Zhang Q H,Chen F S,Zhang X Y.Analysis of culturable yeast diversity in spontaneously fermented orange wine,orange peel and orangery soil of a Ponkan plantation in China.Annals of Microbiology,2015,65(4):2387-2391.
[8]Pathania N,Kanwar S S,Jhang T,Koundal K R,Sharma T R.Application of different molecular techniques for deciphering genetic diversity among yeast isolates of traditional fermented food products of Western Himalayas.World Journal of Microbiology and Biotechnology,2010,26(9):1539-1547.
[9]Oguntoyinbo F A.Culture-independent analysis for determination of yeast diversity during solid substrate fermentation of grated cassava for gari production.World Journal of Microbiology and Biotechnology,2011,27(10):2461-2465.
[10]Haque M A,Seo W T,Hwang C E,Lee H Y,Ahn M J,Cho K M.Culture-independent analysis of yeast diversity in Korean traditional fermented soybean foods(doenjang and kanjang)based on 26S rRNA sequence.Journal of the Korean Society for Applied Biological Chemistry,2015,58(3):377-385.
[11]Rosa C A,Resende M A,Barbosa F A R,Morais P B,Franzot S P.Yeast diversity in a mesotrophic lake on the karstic plateau of Lagoa Santa,MG-Brazil.Hydrobiologia,1995,308(2):103-108.
[12]Yurkov A M,Wehde T,Federici J,Sch?fer A M,Ebinghaus M,Lotze-Engelhard S,Mittelbach M,Prior R,Richter C,R9hl O,Begerow D.Yeast diversity and species recovery rates from beech forest soils.Mycological Progress,2016,15(8):845-859.
[13]Pozo M I,Herrera C M,Bazaga P.Species richness of yeast communities in floral nectar of southern Spanish plants.Microbial Ecology,2011,61(1):82-91.
[14]Kachalkin A V,Yurkov A M.Yeast communities in Sphagnum phyllosphere along the temperature-moisture ecocline in the boreal forest-swamp ecosystem and description of Candida sphagnicola sp.nov.Antonie van Leeuwenhoek,2012,102(1):29-43.
[15]Nasanit R,Jaibangyang S,Tantirungkij M,Limtong S.Yeast diversity and novel yeast D1/D2 sequences from corn phylloplane obtained by a culture-independent approach.Antonie van Leeuwenhoek,2016,109(12):1615-1634.
[16]Wang H M,Xu J P,Guo H,Wu J Y,Yi G H,Pei H,Niu L N,Li Y.Patterns of human oral yeast species distribution on Hainan Island in China.Mycopathologia,2013,176(5/6):359-368.
[17]Faria-Ramos I,Neves-Maia J,Ricardo E,Santos-Antunes J,Silva A T,Costa-de-Oliveira S,Cantón E,Rodrigues A G,Pina-Vaz C.Species distribution and in vitro antifungal susceptibility profiles of yeast isolates from invasive infections during a Portuguese multicenter survey.European Journal of Clinical Microbiology&Infectious Diseases,2014,33(12):2241-2247.
[18]Li W,Hu Y A,Li F Q,Shi L N,Shao H F,Huang M,Wang Y,Han D D,Liao H,Ma C F,Zhang G Y.Distribution of Yeast Isolates from Invasive Infections and Their in vitro susceptibility to antifungal agents:evidence from 299 cases in a 3-year(2010 to 2012)surveillance study.Mycopathologia,2015,179(5/6):397-405.
[19]Gadanho M,Libkind D,Sampaio J.Yeast diversity in the extreme acidic environments of the Iberian pyrite belt.Microbial Ecology,2006,52(3):552-563.
[20]Gümral R,Tümg?r A,Sara?l?M A,Y?ld?ranST,Ilkit M,de Hoog G S.Black yeast diversity on creosoted railway sleepers changes with ambient climatic conditions.Microbial Ecology,2014,68(4):699-707.
[21]Nakase T,Takashima M.A simple procedure for the high frequency isolation of new taxa of ballistosporous yeasts living on the surface of plants.RIKEN Review,1993,3:33-34.
[22]Makimura K,Murayama S Y,Yamaguchi H.Detection of a wide range of medically important fungi by the polymerase chain reaction.Journal of Medical Microbiology,1994,40(5):358-364.
[23]Kurtzman C P,Robnett C J.Identification of clinically important ascomycetous yeasts based on nucleotide divergence in the 5'end of the largesubunit(26S)ribosomal DNA gene.Journal of Clinical Microbiology,1997,35(5):1216-1223.
[24]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.Nucleic Acids Research,1997,25(24):4876-4882.
[25]Saitou N,Nei M.The neighbor-joining method:a new method for reconstructing phylogenetic trees.Molecular Biology and Evolution,1987,4(4):406-425.
[26]Page R D M.Tree View:an application to display phylogenetic trees on personal computers.Bioinformatics,1996,12(4):357-358.
[27]Kurtzman C P,Robnett C J.Identification and phylogeny of ascomycetous yeasts from analysis of nuclear large subunit(26S)ribosomal DNA partial sequences.Antonie van Leeuwenhoek,1998,73(4):331-371.
[28]Fell J W,Boekhout T,Fonseca A,Scorzetti G,Statzell-Tallman A.Biodiversity and systematics of basidiomycetous yeasts as determined by largesubunit r DNA D1/D2 domain sequence analysis.International Journal of Systematic and Evolutionary Microbiology,2000,50(3):1351-1371.
[29]Scorzetti G,Fell J W,Fonseca A,Statzell-Tallman A.Systematics of basidiomycetous yeasts:a comparison of large subunit D1/D2 and internal transcribed spacer r DNA regions.FEMS Yeast Research,2002,2(4):495-571.
[30]Dong L,Zhang J,Sun Y,Liu Y,Zhang Y Y,Zheng G M.Phylogeographic patterns and conservation units of a vulnerable species,Cabot's tragopan(Tragopan caboti),endemic to Southeast China.Conservation Genetics,2010,11(6):2231-2242.
[31]Glushakova A M,Chernov I Y.Seasonal dynamics of the structure of epiphytic yeast communities.Microbiology,2010,79(6):830-839.
[32]Han P J,Li A H,Wang Q M,Bai F Y.Ballistosporomyces changbaiensis sp.nov.and Ballistosporomyces bomiensis sp.nov.,two novel species isolated from shrub plant leaves.Antonie van Leeuwenhoek,2016,109(7):965-970.
[33]Glushakova A M,Chernov I Y.Seasonal dynamics in a yeast population on leaves of the common wood sorrel Oxalis acetosella L.Microbiology,2004,73(2):184-188.
[34]Liu X Z,Wang Q M,G9ker M,Groenewald M,Kachalkin A V,Lumbsch H T,Millanes A M,Wedin M,Yurkov A M,Boekhout T,Bai F Y.Towards an integrated phylogenetic classification of the Tremellomycetes.Studies in Mycology,2015,81:85-147.
[35]Wang Q M,Begerow D,Groenewald M,Liu X Z,Theelen B,Bai F Y,Boekhout T.Multigene phylogeny and taxonomic revision of yeasts and related fungi in the Ustilaginomycotina.Studies in Mycology,2015,81:55-83.
[36]Wang Q M,Yurkov A M,G9ker M,Lumbsch H T,Leavitt S D,Groenewald M,Theelen B,Liu X Z,Boekhout T,Bai F Y.Phylogenetic classification of yeasts and related taxa within Pucciniomycotina.Studies in Mycology,2015,81:149-189.
[37]Hamamoto M,Nakase T.Ballistosporous yeasts found on the surface of plant materials collected in New Zealand.Antonie van Leeuwenhoek,1996,69(3):279-291.
[38]Wang Q M,Bai F Y.Molecular phylogeny of basidiomycetous yeasts in the Cryptococcus luteolus lineage(Tremellales)based on nuclear rRNA and mitochondrial cytochrome b gene sequence analyses:proposal of Derxomyces gen.nov.and Hannaella gen.nov.,and description of eight novel Derxomyces species.FEMS Yeast Research,2008,8(5):799-814.
[39]Smith E P,van Belle G.Nonparametric estimation of species richness.Biometrics,1984,40(1):119-129.