湖光岩玛珥湖浮游细菌与古菌多样性的研究
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摘要
浮游细菌在水生生态系统中分布广,数量巨大,具有丰富的遗传多样性和适应复杂环境的机制,其群落组成和结构的变化在生态系统中具有重要的功能。本论文基于培养与不依赖于培养的方法,对湖光岩采集来的水样分别通过两类通用培养基的纯系培养,结合BOX-PCR带型分析技术,以及通过构建了环境水样总DNA的10个主要微生物门类的文库,阐明了这一典型玛珥湖水层中的古菌多样性和浮游细菌多样性及其季节上的变化。
我们选取了湖光岩湖体的中心点水层(水体最深位点之一)作为研究对象的典型区域,分别在夏季和冬季各采集了1m、5m及13m的水样进行浮游细菌多样性的分析。首先,通过R2A培养基和过滤湖水配制的琼脂培养基培养分离各水层浮游细菌,共获得了340株细菌。通过碱裂解法提取DNA,并进行BOX-PCR反应及手动带型分析筛选差异条带的菌株;以PCR扩增获取筛选的细菌16S rRNA基因用于测序分析。RDP数据分析结果显示,340株菌株分属于变形菌门、放线菌门、厚壁菌门、拟杆菌门等几类湖泊中常出现的主要门类。其中变形菌门虽然跟其他各类细菌一样,在不同季节、水层及培养基上占有的比例不一致,但一般都是处于优势地位,占据着总细菌数的76%以上。另外,拟杆菌在分析的结果上,只出现在冬季的样品中,其数量占总细菌数不大。纯系菌株的16S rRNA基因经BLAST比对后的结果显示,纯系中的各类细菌16S rRNA的同源性大多大于90%,除了湖水配置的培养基分离的夏季水样中Shui5-7、Shui5-12、Shui13-6、Shui13-11和冬季水样中dS1-1、dS5-26,以及R2A培养基上分离的夏季水样中R13-5、R13-7和冬季水样中dR1-11,dR1-21,它们的同源性都不大于90%,可能代表至今尚未认知的新类群。
其次,在上述基于培养的方法研究浮游细菌多样性的基础上,进一步通过不依赖培养的方法,通过10个门类的通用引物对构建了湖光岩水体中古菌、细菌、放线菌、厚壁菌、蓝细菌、浮霉菌、拟杆菌、α-变形菌、β-变形菌和γ-变形菌16S rRNA基因的10个克隆文库。这一方面揭示出了与纯系分离结果不一样的特点,比如在结构上与纯系分离菌株相比,古菌和浮霉菌基本没在两类通用培养基上分离获得,而仅有在构建的文库中出现;另一方面也克隆出了相当一部分对应科属的新序列。这类序列在不同的文库中,比例不同,分别为:占古菌的6/30,占细菌的8/30,占放线菌的2/30,占厚壁菌的2/30,占浮霉菌的23/30,占拟杆菌的29/30,蓝细菌占3/30,占α-变形菌的8/30,占β-变形菌的11/31,占γ-变形菌的27/31。
Numerous planktonic bacteria are widely distributed in aquatic ecosystems. They have high genetic diversity and the capacity of living in almost complex environment on earth. Their community structures play an important role in ecosystem cycles. Here we based on the culture-independent and culture-dependent methods, aiming to study the planktonic bacterial diversity in a typical Maar lake-Hugangyan and furthermore, address its seasonal changes.Our work incledes the analysis of bacteria isolations, culturing by two types of common medium, and the construction of 10 gene clone libraries, being constructed with the lake's main microbial communities. As for the analysis of isolations, we use the BOX-PCR technique to type the various ones.
First, we chose the center vertical watermass of Lake Huguangyan, one of the deepest site, as its typical region and investigated the microbial diversities on 1m, 5m and 13m water layers, respectively, in summer and winter. First of all, 340 bacterial strains were obtained by culturing on R2A medium and agar medium that were prepared by filtered lake water and agar. Then,for the analysis of BOX-PCR we extracted the DNA from each strain by alkaline lysis. Next, strains were distinguished by manual screening on different bands. Later, RDP data analysis showed that 340 strains belonged to Proteobacteria, Actinobacteria, Firmicutes, Bacteroides, which are the main microbial categories in lakes. We found that the proportion of types of bacteria was inconsistent in different seasons, water levels and mediums. But at the same time Proteobacteria is constantly as a superior, occupying more than 76% of the total number of bacteria. In addition, the results of the analysis reveal that Bacteroides was only found in winter samples. The data by BLAST comparison of 16S rRNA genes showed that various types of bacterial 16S rRNA homology are above 90%, excepting the strains of the summer sample from agar medium and R2A medium:Shui5-7, Shui5-12, Shui13-6, Shui13-11, R13-5, R13-7 and dS1-1, dS5-26, dR1-11, dR1-21 in winter. They are not exceeding 90% homology, which may represent a new species in the correspondent categories.
Secondly, in addition to the analysis of bacterial diversity by culture-dependent methods, we also employed culture-independent methods to study the diversity. By using 10 categories of universal primers, we constructed 10 clone libraries of 16S rRNA, including Archaea, Bacteria, Actinomycetes, Firmicutes, Cyanobacteria, Bacteroides, Planctomycetes,α-Proteobacteria,β-Proteobacteria andγ-Proteobacteria. The results revealed different characteristics with the above culturing results. And sequences of Archaea and Planctomycetes were only cloned in these libraries. Besides, there are some more new sequences of other categories from the libraries. The value is 6/30 in Archaea, 8/30 in Bacteria, 2/30 in Actinomycetes, 2/30 in Firmicutes, 23/30 in Planctomycetes, 29/30 in Bacteroides, 3/30 in Cyanobacteria, 8/30 inα-Proteobacteria, 11/31 inβ-Proteobacteria, 27/31 inγ-Proteobacteria, respectively.
我们选取了湖光岩湖体的中心点水层(水体最深位点之一)作为研究对象的典型区域,分别在夏季和冬季各采集了1m、5m及13m的水样进行浮游细菌多样性的分析。首先,通过R2A培养基和过滤湖水配制的琼脂培养基培养分离各水层浮游细菌,共获得了340株细菌。通过碱裂解法提取DNA,并进行BOX-PCR反应及手动带型分析筛选差异条带的菌株;以PCR扩增获取筛选的细菌16S rRNA基因用于测序分析。RDP数据分析结果显示,340株菌株分属于变形菌门、放线菌门、厚壁菌门、拟杆菌门等几类湖泊中常出现的主要门类。其中变形菌门虽然跟其他各类细菌一样,在不同季节、水层及培养基上占有的比例不一致,但一般都是处于优势地位,占据着总细菌数的76%以上。另外,拟杆菌在分析的结果上,只出现在冬季的样品中,其数量占总细菌数不大。纯系菌株的16S rRNA基因经BLAST比对后的结果显示,纯系中的各类细菌16S rRNA的同源性大多大于90%,除了湖水配置的培养基分离的夏季水样中Shui5-7、Shui5-12、Shui13-6、Shui13-11和冬季水样中dS1-1、dS5-26,以及R2A培养基上分离的夏季水样中R13-5、R13-7和冬季水样中dR1-11,dR1-21,它们的同源性都不大于90%,可能代表至今尚未认知的新类群。
其次,在上述基于培养的方法研究浮游细菌多样性的基础上,进一步通过不依赖培养的方法,通过10个门类的通用引物对构建了湖光岩水体中古菌、细菌、放线菌、厚壁菌、蓝细菌、浮霉菌、拟杆菌、α-变形菌、β-变形菌和γ-变形菌16S rRNA基因的10个克隆文库。这一方面揭示出了与纯系分离结果不一样的特点,比如在结构上与纯系分离菌株相比,古菌和浮霉菌基本没在两类通用培养基上分离获得,而仅有在构建的文库中出现;另一方面也克隆出了相当一部分对应科属的新序列。这类序列在不同的文库中,比例不同,分别为:占古菌的6/30,占细菌的8/30,占放线菌的2/30,占厚壁菌的2/30,占浮霉菌的23/30,占拟杆菌的29/30,蓝细菌占3/30,占α-变形菌的8/30,占β-变形菌的11/31,占γ-变形菌的27/31。
Numerous planktonic bacteria are widely distributed in aquatic ecosystems. They have high genetic diversity and the capacity of living in almost complex environment on earth. Their community structures play an important role in ecosystem cycles. Here we based on the culture-independent and culture-dependent methods, aiming to study the planktonic bacterial diversity in a typical Maar lake-Hugangyan and furthermore, address its seasonal changes.Our work incledes the analysis of bacteria isolations, culturing by two types of common medium, and the construction of 10 gene clone libraries, being constructed with the lake's main microbial communities. As for the analysis of isolations, we use the BOX-PCR technique to type the various ones.
First, we chose the center vertical watermass of Lake Huguangyan, one of the deepest site, as its typical region and investigated the microbial diversities on 1m, 5m and 13m water layers, respectively, in summer and winter. First of all, 340 bacterial strains were obtained by culturing on R2A medium and agar medium that were prepared by filtered lake water and agar. Then,for the analysis of BOX-PCR we extracted the DNA from each strain by alkaline lysis. Next, strains were distinguished by manual screening on different bands. Later, RDP data analysis showed that 340 strains belonged to Proteobacteria, Actinobacteria, Firmicutes, Bacteroides, which are the main microbial categories in lakes. We found that the proportion of types of bacteria was inconsistent in different seasons, water levels and mediums. But at the same time Proteobacteria is constantly as a superior, occupying more than 76% of the total number of bacteria. In addition, the results of the analysis reveal that Bacteroides was only found in winter samples. The data by BLAST comparison of 16S rRNA genes showed that various types of bacterial 16S rRNA homology are above 90%, excepting the strains of the summer sample from agar medium and R2A medium:Shui5-7, Shui5-12, Shui13-6, Shui13-11, R13-5, R13-7 and dS1-1, dS5-26, dR1-11, dR1-21 in winter. They are not exceeding 90% homology, which may represent a new species in the correspondent categories.
Secondly, in addition to the analysis of bacterial diversity by culture-dependent methods, we also employed culture-independent methods to study the diversity. By using 10 categories of universal primers, we constructed 10 clone libraries of 16S rRNA, including Archaea, Bacteria, Actinomycetes, Firmicutes, Cyanobacteria, Bacteroides, Planctomycetes,α-Proteobacteria,β-Proteobacteria andγ-Proteobacteria. The results revealed different characteristics with the above culturing results. And sequences of Archaea and Planctomycetes were only cloned in these libraries. Besides, there are some more new sequences of other categories from the libraries. The value is 6/30 in Archaea, 8/30 in Bacteria, 2/30 in Actinomycetes, 2/30 in Firmicutes, 23/30 in Planctomycetes, 29/30 in Bacteroides, 3/30 in Cyanobacteria, 8/30 inα-Proteobacteria, 11/31 inβ-Proteobacteria, 27/31 inγ-Proteobacteria, respectively.
引文
[1]王文远,刘嘉麒.玛珥湖与过去全球变化研究[J].地球科学进展,2000,15(5).
[2]刘嘉麒等,中国玛珥湖的时空分布与地质特征[J].第四纪研究,2000,20(1).
[3]Buchel G.Maars of the Westeifil[J].Lecture Notes in Earth Sciences,1993.
[4]Zolitschka B,Negendank J F W. High-resolution records from European Lakes [J].Quaternary Science Reviews.1999,18:885-888.
[5]Lorenz V. On the formation of maar[J]. Bulletin Volcanologique. 1973, 37:183-204.
[6]Nies DH,Silver S,Bacterial Transition Metal Homeostasis In Molecular Microbiology of Heavy Metals[J].Berlin Heidelberg:Springer-Verlag,2007,117-142.
[7]Bruins MR,Kapil S,Oehme FW.Environment Ecotoxicology and Environmental Safety, 2000,45:198-207.
[8]Binkley D.Interaction of site fertility and red alderon ecosysten production in Douglasfir plantation[J].Forest Ecology and Management,1983,5(1):215-227.
[9]Dazzo F B,Hubbefl DH.Cross-reactive antigens and leetin as determinants of symbiotic specificity in the Rhizobium-clover associatiorl.[J].Microbiol,1975,30(2):1017- 1033.
[10]Vitousek P M,Walker L R,Whiteaker L D,et a1.Biological invasion by Myricafaya alters ecosystem development in Hawaii[J].Science,1987.
[11]Wright R F,Schindler DW.Interaction of acid rain and global changes:effects on terrestrial and aquatic ecosystems [J].Water Airand Soil Pollution,1995,8(2):89-99.
[12]韩兴国,李凌浩,黄建辉,物地球化学循环[M],北京:高教出版社,1999.
[13]李慧蓉.生物监测技术及其研究进展[J].江苏石油化工学院学报.2002,(2).
[14]张春扬,益生菌剂作用机理的研究进展[J].预防兽医学进展,2000,2(4).
[15]NIJU N.ROYC HOUD HURY P K,SRIVASTAVE A.L-(+)lactic acid fermentation and its product polymerization [J].Electron J Biotechn,2004,(8):167- 179.
[16]DATTA R.Technological and economical potential of polylactic acid and lactic acid derivatives [J].FEMS Microbiol Rev,1995,16:221-231.
[17]杨显万,沈庆峰,郭玉霞.微生物湿法冶金[M].北京:冶金工业出版社,2003(09).
[18]邱木清,张卫民.微生物技术在矿产资源利用与环保中的应用[J].矿产保护与利用.2003(6).
[19]徐家振,哲男.金属冶金中的微生物技术[J].有色矿冶,2001(2):31-34.
[20]刘乐然,沿海微生物资源开发利用研究进展[J].河北渔业,2003,4.
[21]康德灿,开发微生物资源创造新型保健食品[J].食品研究与开发, 2001,22(3).
[22]Lindstrêm ES.Bacteriop lankt on community composition in five lakes differing introphic status and humic content[J].Microb Ecol,2000,40:104-113.
[23]Vander G K,Sabbe L K,Meester D,Vloemans N,Zwart G,Gillis M,Vyverman W.Contr- asting bacterioplankt on community composition and seasonal dynamics into neighbouring hypertrophic freshwater lakes [J].Environ Microbiol, 2001, 3: 680-690.
[24]Yannarell AC,Kent AD,Lauster G L,Kratz T K.Temporal patterns in bacterial commun- ities in three temperate lakes of different tr ophic status[J].Microb Ecol,2003, 46: 391-405.
[25]Shaheen B. Humayoun,N. B., et al.Depth Distribution of Microbial Diversity in Mono Lake,a Meromictic Soda Lake in California[J].Appl. Environ. Microbiol.,2003,69(2): 1030 -1042.
[26]Cowan, D.A., Microbial genomes-the untapped resource[J].Trends in Biotechnology, 2000. 18(1),18:14-16.
[27]Boone D R,Castenho1z R W,Garrity G M.BergeY’s Manua1 of Systematic Bacteriolo- gy (Vol.1) [M].New York:Springer,2001.
[28]Regan J.M,Harrington G.W.DiverSity of Nitrifying Bacteria in Fu11 sca1e Ch1oram- inated Distribution SyStems[J].Water ReSearch,2003,37(1):197-205.
[29]Ward DM,Weller R,Bateson MM.16S rRNA sequences reveal numerous uncultured microorganisms in a naturalcommunity[J].Nature,1990,345:3673 -3682.
[30]Streit W R,Schmitz R A.Metagenomies:the key to the uncultured micorbes.Current opinion in Microbiol,2004,7:492-498.
[31]孙鑫鑫,刘惠荣等.乌梁素海富营养化湖区浮游细菌多样性及系统发育分析[J].生物多样性.2009,17(5):490-498.
[32]刘晓波等,青藏高原纳木错湖细菌群落特征及其与高山湖泊的对比[J].冰川冻土.2008.30(6).
[33]孙超,吴敏,史锋.新疆两湖细菌的非培多样性研究[J].浙江大学学报(理学版),2008,35(4).
[34]吴鑫,奚万艳,杨虹,太湖梅梁湾冬季浮游细菌的多样性[J].生态学杂志2006,25(10):1196-1200.
[35]吴根福.杭州西湖水域微生物结构与功能的研究[D].浙江大学,2004.
[36]HideyukiTamaki,et al.Comparative Analysis of Bacterial Diversityin Freshwater Sedi- ment of a Shallow Eutrophic Lake by Molecular and Improved Cultivation-Based Techni- Ques[J].Appl. Environ. Microbiol,2005,4:2162-2169.
[37]KathleenA.Page,et al.Representative Freshwater Bacterioplankton Isolated from Crater Lake,Oregon[J].App. Environ. Microbiol,2004,70(11):6542-6550.
[38]Jakob P,et al.Seasonal Community and Population Dynamics of Pelagic Bactria and Archaea in a High Mountain Lake[J].Appl. Environ. Microbiol,1998,64(11): 4299-4306.
[39]Gl?ckner F O,et al.Comprative 16S rRNA Analysis of Lake Bacterioplankton Reveals Globally Distributed Phylogenetic Clusters Including an Abundant Group of Actinobacter- ia[J].Appl. Environ. Microbiol ,2000, 66(11):5033-5065.
[40]Takai K, Moser DP, Deflaun M, et al. Archaeal diversity in waters from deep south African gold mines[J]. Appl. Environ. Microbiol, 2001, 67(12): 5750?5760.
[41]Anne-C.Lehours,Corinne Bardot,et a1.Anaerobic Microbial Communities in Lake Pav- in,a Unique Meromictic Lake in France[J].Appl.Environ.Microbiol,2005,71(11): 7389- 7400.
[42]Julia I.K Schwarz,Wemer Eckea,et a1.Systematic and Applied Microbiology, 2007, 30:239-254.
[43]Aijaz Ahmad Wani,Venkata Prasad Surakasi,et a1.Research in Microbiology, 2006, 157:928-937.
[44]Santini J M, S1y LI, Schnagl R D et a1.A New Chemo1itho-autotrophie Arsenite- oxidizing Bacterium Iso1ated from a Go1d Mine:Phy1ogenetie,Physiological,and Pre1im- inary Biochemical Studies[J].App1. Environ. Microbiol,2006,66:92-97.
[45]Coates J D,Co1e K A.Diversit Y and Ubiquity of Bacteria Capab1e of Uti1izing Humic Substanees as E1eetron Donors for Anaerobic Respiration[J].App1. Environ. Microbio1, 2002, 68:2445 -2452.
[46]周晓杨等,分子生物学方法在微生物生态学研究中的应用[J].重庆文理学院学报(自然科学版),2007,26(4).
[47]Muyzer G,de Weel E C,Uittrlindern A G.Profiling of complex microbial populations by denaturing gradient gelelectrophoresis analysis of polymerase chain reacti on amplified genes coding for 16S rRNA [J].Appl. Environ. Microbiol,1993,59(3):695-700.
[48]张旋,闫志勇,王斌.UP - PCR结合RFLP快速检测体液标本中病原菌的初步研究和应用[J].首都医药,2003,10:49-51.
[49]Vincent D,Roy D,Mondon F,et al.Characterizati on of bifidobacteria by random DNA amp lification [J].International Journal of Food Microbiology,1998,43(3):185-193.
[50]Hopkins KL,DesaiM,Frost J A,et al.Fluorescent amplified fragment length polymor- phism genotyping of Campylobater jejuni and Campylobacter coli strains and relationship with host specificity,serotyping and phage typing[J].J Clin Microbiol. 2004 Jan;42(1):229- 35.
[51]Vaisanen, R.K., Roberts, M.S., Garland, J.L., Frey, S.D., Dawson, L.A.,Physiological and molecular characterization of microbial communities associated with different water- stable aggregate size fractions[J].Soil Biology and Biochemistry 200537, 2007–2016.
[52]王岩等.PCR-SSCP技术在微生物群落多态性分析中的应用进展[J].生物技术2009,19(3).
[53]李冰冰等.FISH技术及其在环境微生物监测中的应用[J].生物技术.2007,17(5).
[54]PRISCILLAE DOMBEK et al.Use of Repetitive DNA Sequences and the PCR To Differentiate Escherichia coli Isolates from Human and Animal Sources[J].Appl..environ. Microbiol,2000,2572-2577.
[55]杨凤环等,BOX.PCR技术在微生物多样性研究中的应用[J].微生物学通.2008,3 5(8):1282-1286.
[56]Thearith Koeuth,et al,Differential Subsequence Conservation of Interspersed Repetit- ive Streptococcus pneumoniae BOX Elements in Diverse Bacteria[J].Genome Research, 1995(5): 408-418.
[57]Marta Tacao,et al.BOX-PCR is an adequate tool for typing Aeromonas spp. [J].Antoni- evan Leeuwenhoek.2005,88:173-179.
[58]Yasuhiro Oda, et al.Genotypic and Phenotypic Diversity within Species of Purple Nonsulfur Bacteria Isolated from Aquatic Sediments[J].Appl.environ.Microbiol, 2000, 3467-3477.
[59]张才学等,湖光岩玛珥湖的浮游植物[J].水生生物学报.2008,32(5).
[60]刘颖等,湛江湖光岩土壤放线菌分离与抗菌活性测定[J].热带农业工程. 2009,33(1).
[61]丁燏,简纪常,吴灶和等,湖光岩拮抗菌的研究[J].海洋与湖沼.2009,40(3).
[62]顾孔珍,用R2A养基提高饮用水中细菌总数检出率[J],净水技术,2004,23(1).
[63]O’Sullivan L A, Fuller K E et al.Distribution and culturability of the uncultivated 'AGG58 cluster' of the Bacteriodetes phylum in aquatic environments[J]. FEMS Microbiol Ecol 2004, 47:359-370.
[64]金冬雁,黎孟枫等译.分子克隆实验指南[M].第二版.北京:科学出版社,49-55.
[65]白晓慧等,两种培养基对饮水机出水中细菌总数的测定效果比较[J].中国环境监测.2006,22(6).
[66]赵兴青等,基于不同引物的湖泊沉积物中细菌群落结构多样性的比较研究[J].南京大学学报(自然科学)[J].2008,44(3).
[2]刘嘉麒等,中国玛珥湖的时空分布与地质特征[J].第四纪研究,2000,20(1).
[3]Buchel G.Maars of the Westeifil[J].Lecture Notes in Earth Sciences,1993.
[4]Zolitschka B,Negendank J F W. High-resolution records from European Lakes [J].Quaternary Science Reviews.1999,18:885-888.
[5]Lorenz V. On the formation of maar[J]. Bulletin Volcanologique. 1973, 37:183-204.
[6]Nies DH,Silver S,Bacterial Transition Metal Homeostasis In Molecular Microbiology of Heavy Metals[J].Berlin Heidelberg:Springer-Verlag,2007,117-142.
[7]Bruins MR,Kapil S,Oehme FW.Environment Ecotoxicology and Environmental Safety, 2000,45:198-207.
[8]Binkley D.Interaction of site fertility and red alderon ecosysten production in Douglasfir plantation[J].Forest Ecology and Management,1983,5(1):215-227.
[9]Dazzo F B,Hubbefl DH.Cross-reactive antigens and leetin as determinants of symbiotic specificity in the Rhizobium-clover associatiorl.[J].Microbiol,1975,30(2):1017- 1033.
[10]Vitousek P M,Walker L R,Whiteaker L D,et a1.Biological invasion by Myricafaya alters ecosystem development in Hawaii[J].Science,1987.
[11]Wright R F,Schindler DW.Interaction of acid rain and global changes:effects on terrestrial and aquatic ecosystems [J].Water Airand Soil Pollution,1995,8(2):89-99.
[12]韩兴国,李凌浩,黄建辉,物地球化学循环[M],北京:高教出版社,1999.
[13]李慧蓉.生物监测技术及其研究进展[J].江苏石油化工学院学报.2002,(2).
[14]张春扬,益生菌剂作用机理的研究进展[J].预防兽医学进展,2000,2(4).
[15]NIJU N.ROYC HOUD HURY P K,SRIVASTAVE A.L-(+)lactic acid fermentation and its product polymerization [J].Electron J Biotechn,2004,(8):167- 179.
[16]DATTA R.Technological and economical potential of polylactic acid and lactic acid derivatives [J].FEMS Microbiol Rev,1995,16:221-231.
[17]杨显万,沈庆峰,郭玉霞.微生物湿法冶金[M].北京:冶金工业出版社,2003(09).
[18]邱木清,张卫民.微生物技术在矿产资源利用与环保中的应用[J].矿产保护与利用.2003(6).
[19]徐家振,哲男.金属冶金中的微生物技术[J].有色矿冶,2001(2):31-34.
[20]刘乐然,沿海微生物资源开发利用研究进展[J].河北渔业,2003,4.
[21]康德灿,开发微生物资源创造新型保健食品[J].食品研究与开发, 2001,22(3).
[22]Lindstrêm ES.Bacteriop lankt on community composition in five lakes differing introphic status and humic content[J].Microb Ecol,2000,40:104-113.
[23]Vander G K,Sabbe L K,Meester D,Vloemans N,Zwart G,Gillis M,Vyverman W.Contr- asting bacterioplankt on community composition and seasonal dynamics into neighbouring hypertrophic freshwater lakes [J].Environ Microbiol, 2001, 3: 680-690.
[24]Yannarell AC,Kent AD,Lauster G L,Kratz T K.Temporal patterns in bacterial commun- ities in three temperate lakes of different tr ophic status[J].Microb Ecol,2003, 46: 391-405.
[25]Shaheen B. Humayoun,N. B., et al.Depth Distribution of Microbial Diversity in Mono Lake,a Meromictic Soda Lake in California[J].Appl. Environ. Microbiol.,2003,69(2): 1030 -1042.
[26]Cowan, D.A., Microbial genomes-the untapped resource[J].Trends in Biotechnology, 2000. 18(1),18:14-16.
[27]Boone D R,Castenho1z R W,Garrity G M.BergeY’s Manua1 of Systematic Bacteriolo- gy (Vol.1) [M].New York:Springer,2001.
[28]Regan J.M,Harrington G.W.DiverSity of Nitrifying Bacteria in Fu11 sca1e Ch1oram- inated Distribution SyStems[J].Water ReSearch,2003,37(1):197-205.
[29]Ward DM,Weller R,Bateson MM.16S rRNA sequences reveal numerous uncultured microorganisms in a naturalcommunity[J].Nature,1990,345:3673 -3682.
[30]Streit W R,Schmitz R A.Metagenomies:the key to the uncultured micorbes.Current opinion in Microbiol,2004,7:492-498.
[31]孙鑫鑫,刘惠荣等.乌梁素海富营养化湖区浮游细菌多样性及系统发育分析[J].生物多样性.2009,17(5):490-498.
[32]刘晓波等,青藏高原纳木错湖细菌群落特征及其与高山湖泊的对比[J].冰川冻土.2008.30(6).
[33]孙超,吴敏,史锋.新疆两湖细菌的非培多样性研究[J].浙江大学学报(理学版),2008,35(4).
[34]吴鑫,奚万艳,杨虹,太湖梅梁湾冬季浮游细菌的多样性[J].生态学杂志2006,25(10):1196-1200.
[35]吴根福.杭州西湖水域微生物结构与功能的研究[D].浙江大学,2004.
[36]HideyukiTamaki,et al.Comparative Analysis of Bacterial Diversityin Freshwater Sedi- ment of a Shallow Eutrophic Lake by Molecular and Improved Cultivation-Based Techni- Ques[J].Appl. Environ. Microbiol,2005,4:2162-2169.
[37]KathleenA.Page,et al.Representative Freshwater Bacterioplankton Isolated from Crater Lake,Oregon[J].App. Environ. Microbiol,2004,70(11):6542-6550.
[38]Jakob P,et al.Seasonal Community and Population Dynamics of Pelagic Bactria and Archaea in a High Mountain Lake[J].Appl. Environ. Microbiol,1998,64(11): 4299-4306.
[39]Gl?ckner F O,et al.Comprative 16S rRNA Analysis of Lake Bacterioplankton Reveals Globally Distributed Phylogenetic Clusters Including an Abundant Group of Actinobacter- ia[J].Appl. Environ. Microbiol ,2000, 66(11):5033-5065.
[40]Takai K, Moser DP, Deflaun M, et al. Archaeal diversity in waters from deep south African gold mines[J]. Appl. Environ. Microbiol, 2001, 67(12): 5750?5760.
[41]Anne-C.Lehours,Corinne Bardot,et a1.Anaerobic Microbial Communities in Lake Pav- in,a Unique Meromictic Lake in France[J].Appl.Environ.Microbiol,2005,71(11): 7389- 7400.
[42]Julia I.K Schwarz,Wemer Eckea,et a1.Systematic and Applied Microbiology, 2007, 30:239-254.
[43]Aijaz Ahmad Wani,Venkata Prasad Surakasi,et a1.Research in Microbiology, 2006, 157:928-937.
[44]Santini J M, S1y LI, Schnagl R D et a1.A New Chemo1itho-autotrophie Arsenite- oxidizing Bacterium Iso1ated from a Go1d Mine:Phy1ogenetie,Physiological,and Pre1im- inary Biochemical Studies[J].App1. Environ. Microbiol,2006,66:92-97.
[45]Coates J D,Co1e K A.Diversit Y and Ubiquity of Bacteria Capab1e of Uti1izing Humic Substanees as E1eetron Donors for Anaerobic Respiration[J].App1. Environ. Microbio1, 2002, 68:2445 -2452.
[46]周晓杨等,分子生物学方法在微生物生态学研究中的应用[J].重庆文理学院学报(自然科学版),2007,26(4).
[47]Muyzer G,de Weel E C,Uittrlindern A G.Profiling of complex microbial populations by denaturing gradient gelelectrophoresis analysis of polymerase chain reacti on amplified genes coding for 16S rRNA [J].Appl. Environ. Microbiol,1993,59(3):695-700.
[48]张旋,闫志勇,王斌.UP - PCR结合RFLP快速检测体液标本中病原菌的初步研究和应用[J].首都医药,2003,10:49-51.
[49]Vincent D,Roy D,Mondon F,et al.Characterizati on of bifidobacteria by random DNA amp lification [J].International Journal of Food Microbiology,1998,43(3):185-193.
[50]Hopkins KL,DesaiM,Frost J A,et al.Fluorescent amplified fragment length polymor- phism genotyping of Campylobater jejuni and Campylobacter coli strains and relationship with host specificity,serotyping and phage typing[J].J Clin Microbiol. 2004 Jan;42(1):229- 35.
[51]Vaisanen, R.K., Roberts, M.S., Garland, J.L., Frey, S.D., Dawson, L.A.,Physiological and molecular characterization of microbial communities associated with different water- stable aggregate size fractions[J].Soil Biology and Biochemistry 200537, 2007–2016.
[52]王岩等.PCR-SSCP技术在微生物群落多态性分析中的应用进展[J].生物技术2009,19(3).
[53]李冰冰等.FISH技术及其在环境微生物监测中的应用[J].生物技术.2007,17(5).
[54]PRISCILLAE DOMBEK et al.Use of Repetitive DNA Sequences and the PCR To Differentiate Escherichia coli Isolates from Human and Animal Sources[J].Appl..environ. Microbiol,2000,2572-2577.
[55]杨凤环等,BOX.PCR技术在微生物多样性研究中的应用[J].微生物学通.2008,3 5(8):1282-1286.
[56]Thearith Koeuth,et al,Differential Subsequence Conservation of Interspersed Repetit- ive Streptococcus pneumoniae BOX Elements in Diverse Bacteria[J].Genome Research, 1995(5): 408-418.
[57]Marta Tacao,et al.BOX-PCR is an adequate tool for typing Aeromonas spp. [J].Antoni- evan Leeuwenhoek.2005,88:173-179.
[58]Yasuhiro Oda, et al.Genotypic and Phenotypic Diversity within Species of Purple Nonsulfur Bacteria Isolated from Aquatic Sediments[J].Appl.environ.Microbiol, 2000, 3467-3477.
[59]张才学等,湖光岩玛珥湖的浮游植物[J].水生生物学报.2008,32(5).
[60]刘颖等,湛江湖光岩土壤放线菌分离与抗菌活性测定[J].热带农业工程. 2009,33(1).
[61]丁燏,简纪常,吴灶和等,湖光岩拮抗菌的研究[J].海洋与湖沼.2009,40(3).
[62]顾孔珍,用R2A养基提高饮用水中细菌总数检出率[J],净水技术,2004,23(1).
[63]O’Sullivan L A, Fuller K E et al.Distribution and culturability of the uncultivated 'AGG58 cluster' of the Bacteriodetes phylum in aquatic environments[J]. FEMS Microbiol Ecol 2004, 47:359-370.
[64]金冬雁,黎孟枫等译.分子克隆实验指南[M].第二版.北京:科学出版社,49-55.
[65]白晓慧等,两种培养基对饮水机出水中细菌总数的测定效果比较[J].中国环境监测.2006,22(6).
[66]赵兴青等,基于不同引物的湖泊沉积物中细菌群落结构多样性的比较研究[J].南京大学学报(自然科学)[J].2008,44(3).
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