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The diversity of denitrifying bacteria in the alpine meadow soil of Sanjiangyuan natural reserve in Tibet Plateau

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Chinese Science Bulletin

Abstract

This is the first time to describe the diversity of denitrifying bacteria in Sanjiangyuan natural reserve in Tibet Plateau by investigating the molecular diversity and phylogenetic analysis of nirK and nosZ genes using PCR-RFLP and sequencing. Four soil samples were collected from alpine meadow communities from over an altitude of 4600 m which had different physicochemical properties by principal component analysis (PCA). For the genes fragment of nirK and nosZ, the diverse PCR products were characterized by cloning, restriction fragment length polymorphism (RFLP) analysis and sequenced. A total of 253 nirK clones and 283 nosZ clones were received in four samples, and 78 operational taxonomic units (OTUSs) of nirK and 120 OTUs of nosZ by the restriction enzymes Mspl and Rsal digested. The analysis of environmental factors showed that altitude and C/N ratio in soil may be the key factors to the denitrifying bacteria community. 36 nirK clones and 17 nosZ clones were sequenced, and their levels of nucleotide identity were from 69% to 98% and 57% to 97%, respectively. The phylogenetic tree was constructed using the Clustal W and Mega softwares, and all the sequenced clones could be subdivided into 4 groups. Some of clone sequences were related to the nirK and nosZ genes belonging to three phylogenetic subdivisions (α-, β-and γ subclasses of the Proteobacteria), while most of the clones were closely related to the genes of the uncultured bacteria. The sequence distributions were not clear relating to the sample sites in the tree.

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References

  1. Firestone M K. Biological denitrification. In: Stevenson F J, ed. Nitrogen in Agricultural Soils. Madison: American Society for Agronomy, 1982. 289–326

    Google Scholar 

  2. Seitzinger S P. Denitrification in aquatic sediments. In: Revsbech N P, Sorensen J. eds. Denitrification in Soil and Sediment, FEMS Symposium. New York: Plenum Press, 1990. 301–322

    Google Scholar 

  3. Conrad R. Soil microorganisms as controllers of atmospheric trace gases (H2, CO, CH4, OCS, N2O and NO). Microbiol Rev, 1996, 60: 609–640

    Google Scholar 

  4. Zehnder A B. Biology of Anaerobic Microorganisms. New York: John Wiley & Sons, Inc, 1988. 179–244

    Google Scholar 

  5. Rosch C, Mergel A, Bothe H. Biodiversity of denitrifying and dinitrogen-fixing bacteria in an acid forest soil, Appl Environ Microbiol, 2002, 68(8): 3818–3829

    Article  Google Scholar 

  6. Braker G, Zhou J Z, Wu L Y, et al. Nitrite reductase genes (nirK and nirS) as functional markers to investigate diversity of denitrifying bacteria in Pacific Northwest marine sediment communities. Appl Environ Microbiol, 2000, 66(5): 2096–2104

    Article  Google Scholar 

  7. Scala D J, Kerkhof L J. Horizontal heterogeneity of denitrifying bacterial communities in marine sediments by terminal restriction fragment length polymorphism analysis. Appl Environ Microbiol, 2000, 66(5): 1980–1986

    Article  Google Scholar 

  8. Liu X D, Tiquia S M, Holguin G, et al. Molecular diversity of denitrifying genes in continental margin sediments within the oxygen deficient zone of the Pacific Coast of Mexico. Appl Environ Microbiol, 2003, 69(6): 3549–3560

    Article  Google Scholar 

  9. Scala D J, Kerkhof L J. Nitrous oxide reductase (nosZ) gene-specific PCR primers for detection of denitrifiers and three nosZ genes from marine sediments. FEMS Microbiol Lett, 1998, 162: 61–68

    Google Scholar 

  10. Borneman J. Culture-independent identification of microorganisms that respond to specified stimuli. Appl Environ Microbiol, 1999, 65(8): 3398–3400

    Google Scholar 

  11. Stres B, Mahne I, Avgustin G, et al. Nitrous oxide reductase (nosZ) gene fragments differ between native and cultivated Michigan soils. Appl Environ Microbiol, 2004, 70(1): 301–309

    Article  Google Scholar 

  12. Robertson G P, Paul E A, Harwood R R. Greenhouse gases in intensive agriculture: contributions of individual gases to the radiative forcing of the atmosphere. Science, 2000, 289: 1922–1925

    Article  Google Scholar 

  13. Li D Q, Li J W. The Biodiversity in Sanjiangyuan Natural Reserve (in Chinese). Beijing: China Science and Technology Press, 2002. 84–85

    Google Scholar 

  14. Bao S D. Soil and Agricultural Chemistry Analysis (in Chinese). Beijing: China Agriculture Press, 1999. 25–150

    Google Scholar 

  15. Richard A H, Qiu X Y, Wu L Y, et al. Simultaneous recovery of RNA and DNA from soils and sediments. Appl Environ Microbiol, 2001, 67(10): 4495–4503

    Article  Google Scholar 

  16. Zhang Y G, Wang H M, Li D Q, et al. Molecular diversity and phylogenetic analysis of nitrogen-fixing (nifH) genes in alp prairie soil of Sanjiangyuan natural reserve. Acta Microbiologica Sinica (in Chinese), 2005, 45(2: 166–171

    Google Scholar 

  17. Braker G, Fesefeldt A, Witzel K P. Development of PCR primer systems for amplification of nitrite reductase genes (nirK and nirS) to detect denitrifying bacteria in environmental samples. Appl Environ Microbiol, 1998, 64(10): 3769–3775

    Google Scholar 

  18. Zhang Y G, Li D Q, Rao L Q et al. Identification of polymorphic microsatellite DNA locus and paternity test of Amur tiger. Acta Zoologica Sinica (in Chinese), 2003, 49(1): 118–123

    Google Scholar 

  19. Casciotti K L, Ward B B. Dissimilatory nitrite reductase genes from autotrophic ammonia-oxidizing bacteria. Appl Environ Microbiol, 2001, 67(5): 2213–2221

    Article  Google Scholar 

  20. Prieme A, Braker G, Tiedje J M. Diversity of nitrite reductase (nirK and nirS) gene fragments in forested upland and wetland soils. Appl Environ Microbiol, 2002, 68(4): 1893–1900

    Article  Google Scholar 

  21. Song B, Ward B B. Nitrite reductase genes in halobenzoate degrading denitrifying bacteria and related species. FEMS Microbiol Ecol, 2002, 43: 349–357

    Google Scholar 

  22. Cavigelli M, Robertson G P. The functional significance of denitrifier community composition in terrestrial ecosystem. Ecology, 2000, 81: 1402–1414

    Article  Google Scholar 

  23. Cavigelli M, Robertson G P. Role of denitrifier diversity in rates of nitrous oxide consumption in a terrestrial ecosystem. Soil Biol Biochem, 2001, 33: 297–310

    Article  Google Scholar 

  24. Muller C, Martin M, Stevens R J, et al. Processes leading to N2O emissions in grassland soil during freezing and thawing. Soil Biol Biochem, 2002, 34: 1325–1331

    Article  Google Scholar 

  25. Rover M, Heinemeyer O, Kaiser E A. Microbial induced nitrous oxide emissions from arable soil during winter Soil. Biol Biochem, 1998, 30: 1859–1865

    Article  Google Scholar 

  26. Wieland G, Neumann R, Backhaus H. Variation of microbial communities in soil, rhizosphere, and rhizoplane in response to crop species, soil type, and crop development. Appl Environ Microbiol, 2001, 67(12): 5849–5854

    Article  Google Scholar 

  27. Zhou J, Xia B, Treves D S, et al. Spatial and resource factors influencing high microbial diversity in soil. Appl Environ Microbiol, 2002, 68(1): 326–334

    Article  Google Scholar 

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Correspondence to Li Diqiang.

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Zhang, Y., Li, D., Wang, H. et al. The diversity of denitrifying bacteria in the alpine meadow soil of Sanjiangyuan natural reserve in Tibet Plateau. CHINESE SCI BULL 51, 1245–1254 (2006). https://doi.org/10.1007/s11434-006-1245-7

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  • DOI: https://doi.org/10.1007/s11434-006-1245-7

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