明永冰川垂直气候带中可培养低温细菌多样性分析
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摘要
本研究对位于云南滇西北的明永冰川地区暖温带、中温带和寒温带三个不同垂直气候带中可培养低温细菌的多样性进行了研究。利用四种不同培养基对该地区可培养低温细菌进行了分离纯化,共得到细菌37 513株,根据菌落形态特征分为了391种,其中LB培养基分离到99种,Organic培养基分离到78种,PSG培养基分离到96种,PYGV培养基分离得到118种,可以看出寡营养培养基PYGV分离得到的细菌种类多于LB和Organnic等富营养培养基,表明PYGV针对冰川地区细菌的分离与鉴定更为合适;通过革兰氏染色和扫描电镜观察表明大部分菌株为革兰阴性杆菌;对已分离得到的优势菌进行了16S rRNA基因测序并构建系统发育树,分析得出:假单胞菌属(Pseudomonas)、耶尔森氏菌属(Yersinia)和黄杆菌属(Flavobacterium)在明永冰川不同垂直气候带上均有分布,其中假单胞菌属最多占据35%;而寡养单胞菌属(Stenotrophomonas)是寒温带上特有的菌属。本研究证明明永冰川地区垂直气候带中可培养低温细菌多样性非常丰富,也为下一步了解这一特殊地理生态环境下微生物的群落演替规律、研究冰川环境中微生物群落如何响应气候变化提供了参考。
In this study, we studied the diversity of culturable low-temperature bacteria in three vertical climatic zones that included warm temperate, middle temperate and cold temperate zones distributed along the Mingyong glacier in northwest of Yunnan province. Using four different media, 37 513 bacterial isolates were obtained.According to the morphological characteristics of isolate colony, 391 bacterial trains form the isolates were identified. Among them, the strains coded No. 99, 78, 96 and 118 were cultured with LB, Organic, PSG and PYGV medium respectively. PYGV medium showed more effective than that of eutrophic LB and Organnic medium in bacteria isolation in this area, which indicated that oligotrophic medium should be suitable to isolate bacteria from the glacier region. Most of isolates were gram-negative bacilli according to the results of Gram staining and scanning electron microscopic observation; parts of dominant bacterial 16 S rRNA sequences were sequenced and the phylogenetic trees were constructed as well. The studies have shown that the species of Pseudomonas, Yersinia and Flavobacterium were presence in all three vertical climatic zones in Mingyong glacier. Among them,Pseudomonas is the dominant population and occupies up to 35%, and Stenotrophomonas is a unique species distributed in cold temperate zone. Through studying the diversity of culturable low-temperature bacteria in Mingyong glacier, it would provide valuable information for the further study of microbial community in this special geographical environment as well as the reference for the understanding of how the microbial community of the glacier environment response to climate changes.
In this study, we studied the diversity of culturable low-temperature bacteria in three vertical climatic zones that included warm temperate, middle temperate and cold temperate zones distributed along the Mingyong glacier in northwest of Yunnan province. Using four different media, 37 513 bacterial isolates were obtained.According to the morphological characteristics of isolate colony, 391 bacterial trains form the isolates were identified. Among them, the strains coded No. 99, 78, 96 and 118 were cultured with LB, Organic, PSG and PYGV medium respectively. PYGV medium showed more effective than that of eutrophic LB and Organnic medium in bacteria isolation in this area, which indicated that oligotrophic medium should be suitable to isolate bacteria from the glacier region. Most of isolates were gram-negative bacilli according to the results of Gram staining and scanning electron microscopic observation; parts of dominant bacterial 16 S rRNA sequences were sequenced and the phylogenetic trees were constructed as well. The studies have shown that the species of Pseudomonas, Yersinia and Flavobacterium were presence in all three vertical climatic zones in Mingyong glacier. Among them,Pseudomonas is the dominant population and occupies up to 35%, and Stenotrophomonas is a unique species distributed in cold temperate zone. Through studying the diversity of culturable low-temperature bacteria in Mingyong glacier, it would provide valuable information for the further study of microbial community in this special geographical environment as well as the reference for the understanding of how the microbial community of the glacier environment response to climate changes.
引文
An esio A.M.,and Laybourn-Parry J.,2012,Glaciers and ice sheets as a biome,Trends Ecol.Evol.,27(4):219-225
Carney K.M.,and Matson P.A.,2005,Plant communities,soi microorganisms,and soil carbon cycling:does altering the world belowground matter to ecosystem functioning,Ecosystems,8(8):928-940
Balkwill D.L.,1989,Numbers,diversity,and morphologica characteristics of aerobic,chemoheterotrophic bacteria in deep subsurface sediments from a site in South Carolina Geomicrobiol.J.,7(1-2):33-52
Doyle S.M.,Montross S.N.,Skidmore M.L.,and Christner B.C.2013,Characterizing microbial diversity and the potentia for metabolic function at-15℃in the basal ice of Taylor Glacier,Antarctica,Biology,2(3):1034-1053
Gu Y.F.,Zheng Y.K.,Penttinen P.,Wang X.B.,and Zhang X.P.2014,Phylogeny of psychrophilic bacteria from peat swamp soil in Zoigêplateau,Shidi Kexue(Wetland Science),12(5)631-637(辜运富,郑有坤,Penttinen P.,王宪斌,张小平2014,若尔盖高原泥炭沼泽土嗜冷细菌系统发育分析,湿地科学,12(5):631-637)
Li H.Y.,2016,Bacterial community diversity in the vertical climate zones of Mingyong Glacier and retreat region,Dessertation for Ph.D.,Kunming University of Science and Technology,Supersivor:Wei Y.L.,pp.31-49(李浩宇,2016,明永冰川及其退却地区垂直气候带的细菌群落多样性研究,博士学位论文,昆明理工大学,导师:魏云林,pp.31-49)
Li M.Y.,Wang J.L.,and Sawut G.,2015,Culturable bacterial diversity in snow,ice and meltwater of the Yangbark Glacier,Muztag Ata,Bingchuan Dongtu(Journal of Glaciology and Geocryology),37(6):1634-1641(李明源,王继莲,古丽巴哈尔·萨吾提,2015,新疆东帕米尔高原慕士塔格峰洋布拉克冰川雪冰及融水中可培养细菌多样性分析,冰川冻土,37(6):1634-1641)
Liu Y.Q.,Yao T.D.,Jiao N.Z.,Kang S.C.,Huang S.G.,Li Q.,Wang K.J.,and Liu X.B.,2009,Culturable bacteria in glacial meltwater at 6 350 m on the East Rongbuk Glacier,Mount Everest,Extremophiles,13(1):89-99
Li Y.Q.,Zhang C.B.,Zhao H.K.,Tan H.,Fei X.Y.,Zhao L.M.,and Dong Y.S.,2015,Extraction of high purity mitochondrial DNA from Glycine max for genome sequencing,Jiyinzuxue Yu Yingyong Shengwuxue(Genomics and Applied Biology),34(8):1734-1738(李玉秋,张春宝,赵洪锟,谭化,费晓燕,赵丽梅,董英山,2015,适于基因组测序的高纯度的大豆线粒体基因组DNA提取方法,基因组学与应用生物学,34(8):1734-1738)
Lay C.Y.,Mykytczuk N.C.,Niederberger T.D.,Martineau C.,Greer C.W.,and Whyte L.G.,2012,Microbial diversity and activity in hypersaline high Arctic spring channels,Extremophiles,16(2):177-191
Philippot L.,Tscherko D.,Bru D.,and Kandeler E.,2011,Distribution of high bacterial taxa across the chronosequence of two alpine glacier forelands,Microb.Ecol.,61(2):303-312
Simu K.,and Hagstr?m A.,2004,Oligotrophic bacterioplankton with a novel single-cell life strategy,Appl.Environ.Microbiol.,70(4):2445-2251
Steven B.,Pollard W.H.,Greer C.W.,and Whyte L.G.,2008,Microbial diversity and activity through a permafrost/ground ice core profile from the Canadian high Arctic,Environ.Microbiol.,10(12):3388-3403
Christner B.C.,Mosley-Thompson E.,Thompson L.G.,Zagorodnov V.,Sandman K.,and Reeve J.N.,2000,Recovery and identification of viable bacteria immured in Glacial Ice,I-carus,144(2):479-485
Tian W.,Wang L.,Li G.,Li Y.,and Zhang J.B.,2014,Isolation of thermophilic cellulose-decomposing bacteria and cloning and expression of an endoglucanase gene,Nongye Huanjing Kexue Xuebao(Journal of Agro-Environment Science),33(4):788-793(田伟,王磊,李刚,李妍,张纪兵,2014,高温纤维素降解菌的筛选及内切酶基因的克隆表达,农业环境科学学报,33(4):788-793)
Wei Y.L.,Zhou C.T.,Ji X.L.,Lin L.B.,and Jing S.R.,2007,Phylogenetic analysis of several pseudomonas strains isolated from Mingyong glacier,Yunnan Daxue Xuebao(Journal of Yunnan University(Natural Science)),27(S3):408-412(魏云林,周长涛,季秀玲,林连兵,井申荣,2007,明永冰川地区假单胞菌的分离及系统发育分析,云南大学学报(自然科学版),27(S3):408-412)
Xu J.,and Wilkes A.,2004,Biodiversity impact analysis in northwest Yunnan,southwest China,Biodivers.Conser.,13(5)959-983
Zhang S.H.,Hou S.G.,Qin X.,Du W.T.,Li Z.G.,Zhao L.F.,and Liang F.,2014,Progress in research on resources of glacia microbiology,Huanjingkexue Yu Jishu(Environmental Science and Technology),37(12):62-67(张淑红,侯书贵,秦翔,杜文涛,李治国,赵龙飞,梁峰,2014,冰川微生物资源研究进展,环境科学与技术,37(12):62-67)
Zhang Z.,Ou X.,Wang C.,and Wu Y.,2004,Accuracy analysis of vegetation mapping for Meili Snow Mountain area,northwest Yunnan,China,Chinese Journal of Applied Ecology,15(9):1517-1522
Zhang S.H.,Hou S.G.,and Qin D.H.,2007,Advances in researches of microorganisms in glacial snow and ice on the Qinghai-Tibet Plateau,Yingyong Yu Huangjing Shengwu Xuebao(Chinese Journal of Applied and Environmental Biology),13(4):592-596(张淑红,侯书贵,秦大河,2007,青藏高原冰川雪冰微生物研究进展,应用与环境生物学报,13(4):592-596)
Carney K.M.,and Matson P.A.,2005,Plant communities,soi microorganisms,and soil carbon cycling:does altering the world belowground matter to ecosystem functioning,Ecosystems,8(8):928-940
Balkwill D.L.,1989,Numbers,diversity,and morphologica characteristics of aerobic,chemoheterotrophic bacteria in deep subsurface sediments from a site in South Carolina Geomicrobiol.J.,7(1-2):33-52
Doyle S.M.,Montross S.N.,Skidmore M.L.,and Christner B.C.2013,Characterizing microbial diversity and the potentia for metabolic function at-15℃in the basal ice of Taylor Glacier,Antarctica,Biology,2(3):1034-1053
Gu Y.F.,Zheng Y.K.,Penttinen P.,Wang X.B.,and Zhang X.P.2014,Phylogeny of psychrophilic bacteria from peat swamp soil in Zoigêplateau,Shidi Kexue(Wetland Science),12(5)631-637(辜运富,郑有坤,Penttinen P.,王宪斌,张小平2014,若尔盖高原泥炭沼泽土嗜冷细菌系统发育分析,湿地科学,12(5):631-637)
Li H.Y.,2016,Bacterial community diversity in the vertical climate zones of Mingyong Glacier and retreat region,Dessertation for Ph.D.,Kunming University of Science and Technology,Supersivor:Wei Y.L.,pp.31-49(李浩宇,2016,明永冰川及其退却地区垂直气候带的细菌群落多样性研究,博士学位论文,昆明理工大学,导师:魏云林,pp.31-49)
Li M.Y.,Wang J.L.,and Sawut G.,2015,Culturable bacterial diversity in snow,ice and meltwater of the Yangbark Glacier,Muztag Ata,Bingchuan Dongtu(Journal of Glaciology and Geocryology),37(6):1634-1641(李明源,王继莲,古丽巴哈尔·萨吾提,2015,新疆东帕米尔高原慕士塔格峰洋布拉克冰川雪冰及融水中可培养细菌多样性分析,冰川冻土,37(6):1634-1641)
Liu Y.Q.,Yao T.D.,Jiao N.Z.,Kang S.C.,Huang S.G.,Li Q.,Wang K.J.,and Liu X.B.,2009,Culturable bacteria in glacial meltwater at 6 350 m on the East Rongbuk Glacier,Mount Everest,Extremophiles,13(1):89-99
Li Y.Q.,Zhang C.B.,Zhao H.K.,Tan H.,Fei X.Y.,Zhao L.M.,and Dong Y.S.,2015,Extraction of high purity mitochondrial DNA from Glycine max for genome sequencing,Jiyinzuxue Yu Yingyong Shengwuxue(Genomics and Applied Biology),34(8):1734-1738(李玉秋,张春宝,赵洪锟,谭化,费晓燕,赵丽梅,董英山,2015,适于基因组测序的高纯度的大豆线粒体基因组DNA提取方法,基因组学与应用生物学,34(8):1734-1738)
Lay C.Y.,Mykytczuk N.C.,Niederberger T.D.,Martineau C.,Greer C.W.,and Whyte L.G.,2012,Microbial diversity and activity in hypersaline high Arctic spring channels,Extremophiles,16(2):177-191
Philippot L.,Tscherko D.,Bru D.,and Kandeler E.,2011,Distribution of high bacterial taxa across the chronosequence of two alpine glacier forelands,Microb.Ecol.,61(2):303-312
Simu K.,and Hagstr?m A.,2004,Oligotrophic bacterioplankton with a novel single-cell life strategy,Appl.Environ.Microbiol.,70(4):2445-2251
Steven B.,Pollard W.H.,Greer C.W.,and Whyte L.G.,2008,Microbial diversity and activity through a permafrost/ground ice core profile from the Canadian high Arctic,Environ.Microbiol.,10(12):3388-3403
Christner B.C.,Mosley-Thompson E.,Thompson L.G.,Zagorodnov V.,Sandman K.,and Reeve J.N.,2000,Recovery and identification of viable bacteria immured in Glacial Ice,I-carus,144(2):479-485
Tian W.,Wang L.,Li G.,Li Y.,and Zhang J.B.,2014,Isolation of thermophilic cellulose-decomposing bacteria and cloning and expression of an endoglucanase gene,Nongye Huanjing Kexue Xuebao(Journal of Agro-Environment Science),33(4):788-793(田伟,王磊,李刚,李妍,张纪兵,2014,高温纤维素降解菌的筛选及内切酶基因的克隆表达,农业环境科学学报,33(4):788-793)
Wei Y.L.,Zhou C.T.,Ji X.L.,Lin L.B.,and Jing S.R.,2007,Phylogenetic analysis of several pseudomonas strains isolated from Mingyong glacier,Yunnan Daxue Xuebao(Journal of Yunnan University(Natural Science)),27(S3):408-412(魏云林,周长涛,季秀玲,林连兵,井申荣,2007,明永冰川地区假单胞菌的分离及系统发育分析,云南大学学报(自然科学版),27(S3):408-412)
Xu J.,and Wilkes A.,2004,Biodiversity impact analysis in northwest Yunnan,southwest China,Biodivers.Conser.,13(5)959-983
Zhang S.H.,Hou S.G.,Qin X.,Du W.T.,Li Z.G.,Zhao L.F.,and Liang F.,2014,Progress in research on resources of glacia microbiology,Huanjingkexue Yu Jishu(Environmental Science and Technology),37(12):62-67(张淑红,侯书贵,秦翔,杜文涛,李治国,赵龙飞,梁峰,2014,冰川微生物资源研究进展,环境科学与技术,37(12):62-67)
Zhang Z.,Ou X.,Wang C.,and Wu Y.,2004,Accuracy analysis of vegetation mapping for Meili Snow Mountain area,northwest Yunnan,China,Chinese Journal of Applied Ecology,15(9):1517-1522
Zhang S.H.,Hou S.G.,and Qin D.H.,2007,Advances in researches of microorganisms in glacial snow and ice on the Qinghai-Tibet Plateau,Yingyong Yu Huangjing Shengwu Xuebao(Chinese Journal of Applied and Environmental Biology),13(4):592-596(张淑红,侯书贵,秦大河,2007,青藏高原冰川雪冰微生物研究进展,应用与环境生物学报,13(4):592-596)
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