水环境细菌16S rDNA限制性片段长度多型性及群落结构分析

李志岗, 杨官品, 朱艳红

李志岗, 杨官品, 朱艳红. 水环境细菌16S rDNA限制性片段长度多型性及群落结构分析[J]. 水生生物学报, 2001, 25(2): 111-115.
引用本文: 李志岗, 杨官品, 朱艳红. 水环境细菌16S rDNA限制性片段长度多型性及群落结构分析[J]. 水生生物学报, 2001, 25(2): 111-115.
LI Zhi-gang, YANG Guan-pin, ZHU Yan-hong. THE RELATIONSHIP BETWEEN POLYMORPHISM OF BACTERIAL 16S rDNA RESTRICTION FRAGMENT LENGTH AND COMMUNITY STRUCTURE OF AQUATIC BACTERIA[J]. ACTA HYDROBIOLOGICA SINICA, 2001, 25(2): 111-115.
Citation: LI Zhi-gang, YANG Guan-pin, ZHU Yan-hong. THE RELATIONSHIP BETWEEN POLYMORPHISM OF BACTERIAL 16S rDNA RESTRICTION FRAGMENT LENGTH AND COMMUNITY STRUCTURE OF AQUATIC BACTERIA[J]. ACTA HYDROBIOLOGICA SINICA, 2001, 25(2): 111-115.

水环境细菌16S rDNA限制性片段长度多型性及群落结构分析

基金项目: 

武汉市科委晨光计划项目

湖北省自然科学基金资助

THE RELATIONSHIP BETWEEN POLYMORPHISM OF BACTERIAL 16S rDNA RESTRICTION FRAGMENT LENGTH AND COMMUNITY STRUCTURE OF AQUATIC BACTERIA

  • 摘要: 用细菌16S核糖体RNA基因(rDNA)限制性片段长度多型性描述了水环境细菌群落结构.从环境水样中直接分离 DNA,以细菌特异的引物扩增 165 rDNA,构建质粒文库,随机分离重组质粒,用限制性内切酶消化获得 16S rDNA基因型,用基因型的种类及频率描述特定水体生境的细菌群落结构.该方法在分析水体隐含遗传多样性、揭示污染的生物学效应和评价水环境质量等方面具有重要的应用价值.
    Abstract: In this study, a technique describing aquatic bacterial community structure has been developed by using restriction fragment length polymorphism of 16S ribosomal RNA gene(rDNA). The technique includes isolation of DNA directly from lake-water, amplification of rDNA using bacteria-specific primers, construction of plasmid library and digestion of randomly extracted recombinant plasmids with restriction endonuclease. The structure of bacterial community is determined using rDNA genotypes and their frequencies. This method has potentials in analyzing hidden genetic diversity of aquatic environments, tracing biological effectiveness of pollutants and evaluating environmental quality.
  • [1] Amann R I,Ludwig W,Schleifer K H.Phylogenetic identification and in situ detection of individual microbial cells without cultivation [J].Microbiology Reviews.1995,59:143—169[2] Marchesi J R,Sato T,Weightman A J,et al.Design and examination of useful bacterium specific PCR primers that amplify genes coding for bacterial 16S rRNA [J].Appl.Environ.Microbiol.,1998,64:795—799[3] Field K G,Gordon D,Wright T,et al.Diversity and depth specific distribution of SAR 11 cluster rRNA genes from marine planktonic bacteria [J].Appl.Environ.Microbiol.,1997,63:63—70[4] Wise M G,McArther J V,Shimkets L J.Bacterial diversity of a Carolina bay as determined by 16S rRNA gene analysis:Confirmation of novel taxa [J].Appl.Environ.Microbiol.,1997,63:1505—1514[5] Collins M D,East A K.Phylogeny and taxonomy of the food-borne pathogen Clostridium botulinum and its neurotoxins [J].J.of Appl.Mocrobiol.,1998,84:5—17[6] Laguerre G,Allard M R,Revoy F,et al.Rapid identification of rhizobia by restriction fragment length polymorphism analysis of PCR-amplified 16S rRNA genes [J].Appl.Environ.Microbiol.,1994,60:56—63[7] Brunel B,Givaudan A,Lanois A et al.Fast and accurate identification of Xenorhabdus and Photorhabdus species by restriction analysis of PCR-amplified 16S rRNA genes [J].Appl.Environ.Microbiol.,1997,63:574—580[8] 谭志远,陈文新.根瘤菌新类群的全细胞蛋白电泳及16S rDNA全序列分析[J].应用与环境生物学报,1998,4:65—69[9] Britschgi T B,Giovannoni S.Analysis of a natural marine bacterioplankton population by rRNA cloning and sequencing [J].Applied and Environmental Microbiology,1991,57:1707—1713[10] Lane D J,Field K G,Olsen G J,et al.Reverse transcriptase sequencing of ribosomal RNA for phylogenetic analysis [J].Methods Enzymol.,1998,167:138—144[11] 杨官品,朱艳红.土壤细菌基因资源的直接分离:16S核糖体RNA基因模式[J].湖北大学学报,1998,4:383—385[12] Yang G P,Saghai Maroof M A,Zhang Q F,et al.Comparative analysis of microsatellite DNA polymorphism in landraces and cultivars of rice [J].Mol.Gen.Genet.,1994,245:178—194[13] Southern E M.Measurement of DNA length by gel electrophoresis [J].Anal.Biochem.,1979,100:319—323[14] Wang H,Kohalmi S E,et al.An improved method for polymerase chain reaction using whole yeast cells [J].Analytical Biochemistry,1996,237:145—146[15] Redecker D,Thierfelder H,et al.Restriction analysis of PCR-amplified internal transcribed spacers of ribosomal DNA as a tool for species identification in different genera of the order glomals [J].Applied and Environmental Microbiology,1997,63:1756—1761

    Amann R I,Ludwig W,Schleifer K H.Phylogenetic identification and in situ detection of individual microbial cells without cultivation [J].Microbiology Reviews.1995,59:143—169[2] Marchesi J R,Sato T,Weightman A J,et al.Design and examination of useful bacterium specific PCR primers that amplify genes coding for bacterial 16S rRNA [J].Appl.Environ.Microbiol.,1998,64:795—799[3] Field K G,Gordon D,Wright T,et al.Diversity and depth specific distribution of SAR 11 cluster rRNA genes from marine planktonic bacteria [J].Appl.Environ.Microbiol.,1997,63:63—70[4] Wise M G,McArther J V,Shimkets L J.Bacterial diversity of a Carolina bay as determined by 16S rRNA gene analysis:Confirmation of novel taxa [J].Appl.Environ.Microbiol.,1997,63:1505—1514[5] Collins M D,East A K.Phylogeny and taxonomy of the food-borne pathogen Clostridium botulinum and its neurotoxins [J].J.of Appl.Mocrobiol.,1998,84:5—17[6] Laguerre G,Allard M R,Revoy F,et al.Rapid identification of rhizobia by restriction fragment length polymorphism analysis of PCR-amplified 16S rRNA genes [J].Appl.Environ.Microbiol.,1994,60:56—63[7] Brunel B,Givaudan A,Lanois A et al.Fast and accurate identification of Xenorhabdus and Photorhabdus species by restriction analysis of PCR-amplified 16S rRNA genes [J].Appl.Environ.Microbiol.,1997,63:574—580[8] 谭志远,陈文新.根瘤菌新类群的全细胞蛋白电泳及16S rDNA全序列分析[J].应用与环境生物学报,1998,4:65—69[9] Britschgi T B,Giovannoni S.Analysis of a natural marine bacterioplankton population by rRNA cloning and sequencing [J].Applied and Environmental Microbiology,1991,57:1707—1713[10] Lane D J,Field K G,Olsen G J,et al.Reverse transcriptase sequencing of ribosomal RNA for phylogenetic analysis [J].Methods Enzymol.,1998,167:138—144[11] 杨官品,朱艳红.土壤细菌基因资源的直接分离:16S核糖体RNA基因模式[J].湖北大学学报,1998,4:383—385[12] Yang G P,Saghai Maroof M A,Zhang Q F,et al.Comparative analysis of microsatellite DNA polymorphism in landraces and cultivars of rice [J].Mol.Gen.Genet.,1994,245:178—194[13] Southern E M.Measurement of DNA length by gel electrophoresis [J].Anal.Biochem.,1979,100:319—323[14] Wang H,Kohalmi S E,et al.An improved method for polymerase chain reaction using whole yeast cells [J].Analytical Biochemistry,1996,237:145—146[15] Redecker D,Thierfelder H,et al.Restriction analysis of PCR-amplified internal transcribed spacers of ribosomal DNA as a tool for species identification in different genera of the order glomals [J].Applied and Environmental Microbiology,1997,63:1756—1761

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出版历程
  • 收稿日期:  1998-06-19
  • 修回日期:  2000-07-29
  • 发布日期:  2001-03-24

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