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PCR-DGGE Comparison of Bacterial Community Structure in Fresh and Archived Soils Sampled along a Chihuahuan Desert Elevational Gradient

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Abstract

The polymerase chain reaction coupled with denaturing gradient gel electrophoresis (PCR-DGGE) has been used widely to determine species richness and structure of microbial communities in a variety of environments. Researchers commonly archive soil samples after routine chemical or microbial analyses, and applying PCR-DGGE technology to these historical samples offers evaluation of long-term patterns in bacterial species richness and community structure that was not available with previous technology. However, use of PCR-DGGE to analyze microbial communities of archived soils has been largely unexplored. To evaluate the stability of DGGE patterns in archived soils in comparison with fresh soils, fresh and archived soils from five sites along an elevational gradient in the Chihuahuan Desert were compared using PCR-DGGE of 16S rDNA. DNA from all archived samples was extracted reliably, but DNA in archived soils collected from a closed-canopy oak forest site could not be amplified. DNA extraction yields were lower for most archived soils, but minimal changes in bacterial species richness and structure due to archiving were noted in bacterial community profiles from four sites. Use of archived soils to determine long-term changes in bacterial community structure via PCR-DGGE appears to be a viable option for addressing microbial community dynamics for particular ecosystems or landscapes.

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References

  1. Blake GR, Hartage KH (1986) Bulk density. In: Klute A (ed) Methods of soil analysis: part 1: physical and mineralogical methods monograph no. 9. American Society of Agronomy and Soil Science Society of America, Madison, WI, pp 363–375

    Google Scholar 

  2. Boone RD, Grigal DF, Sollins P, Ahrens RJ, Armstrong DE (1999) Soil sampling, preparation, archiving, and quality control. In: Robertson GP, Coleman DC, Bledsoe CS, Sollins P (eds) Standard soil methods for long-term ecological research. Oxford University Press, New York, NY, pp 3–28

    Google Scholar 

  3. Cai P, Huang Q-Y, Zhang X-W (2006) Interactions of DNA with clay minerals and soil colloidal particles and protection against degradation by DNase. Environ Sci Technol 40:2971–2976

    Article  PubMed  CAS  Google Scholar 

  4. Dolfing J, Vos A, Bloem J, Ehlert PA, Naumova NB, Kuikman PJ (2004) Microbial diversity in archived soils. Science 306:813

    Article  PubMed  CAS  Google Scholar 

  5. Dong D, Yan A, Liu H, Zhang X, Xu Y (2006) Removal of humic substances from soil DNA using aluminium sulfate. J Microbiol Methods 66:217–222

    Article  PubMed  CAS  Google Scholar 

  6. Franchi M, Braanti E, Bonzi LM, Orioli PL, Vettori C, Gallori E (1997) Clay-nucleic acid complexes: characteristics and implications for the preservation of genetic material in primeval habitats. Orig Life Evol Biosph 29:297–315

    Article  Google Scholar 

  7. Moon-van der Staay S, Tzeneva VA, van der Staay GWM, de Vos WM, Smidt H, Hackstein JHP (2006) Eukaryotic diversity in historical soil samples. FEMS Microbiol Ecol 57:420–428

    Article  PubMed  CAS  Google Scholar 

  8. Muyzer G, de Waal EC, Uitterlinden AG (1993) Profiling of complex microbial populations by denaturing gradient gel electrophoresis analysis of polymerase chain reaction-amplified genes coding for 16S rRNA. Appl Environ Microbiol 59:695–700

    PubMed  CAS  Google Scholar 

  9. Nunan N, Morgan MA, Herlihy M (1998) Ultraviolet absorbance (280 nm) of compounds released from soil during chloroform fumigation as an estimate of the microbial biomass. Soil Biol Biochem 30:1599–1603

    Article  CAS  Google Scholar 

  10. Robertson GP, Sollins P, Boyd GE, Lajitha K (1999) Exchangeable ions, pH, and cation exchange capacity. In: Robertson GP, Coleman DC, Bledsoe CS, Sollins P (eds) Standard soil methods for long-term ecological research. Oxford University Press, New York, NY, pp 106–114

    Google Scholar 

  11. Sobek EA, Zak JC (2003) The soil FungiLog procedure: method and analytical approaches toward understanding fungal functional diversity. Mycologia 95:590–602

    Article  Google Scholar 

  12. Sollins P, Glassman C, Paul EA, Swanston C, Lajitha K, Heil JW, Elliot ET (1999) Soil carbon and nitrogen: pools and fractions. In: Robertson GP, Coleman DC, Bledsoe CS, Sollins P (eds) Standard soil methods for long-term ecological research. Oxford University Press, New York, NY, pp 89–105

    Google Scholar 

  13. Trevors JT (1996) DNA in soil: adsorption, genetic transformation, molecular evolution and genetic microchip. Antonie Van Leeuwenhoek 70:1–10

    Article  PubMed  CAS  Google Scholar 

  14. Trumbore SE (1993) Comparisons of carbon dynamics in tropical and temperate soils using radiocarbon measurements. Glob Biogeochem Cycles 7:275–290

    Article  CAS  Google Scholar 

  15. Turner AJ (1977) Soil survey of Jeff Davis County, Texas. USDA, Texas, pp 3–12

    Google Scholar 

  16. Tzeneva VA, Li Y, Felske ADM, de Vos WM, Akkermans ADL, Vaughan EE, Smidt H (2004) Development and application of a selective PCR-denaturing gradient gel electrophoresis approach to detect a recently cultivated Bacillus group predominant in soil. Appl Environ Microbiol 70:5801–5809

    Article  PubMed  CAS  Google Scholar 

  17. Van Miegroet H (1995) Inorganic nitrogen determined by laboratory and field extractions of two forest soils. Soil Sci Soc Am J 59:549–553

    Article  Google Scholar 

  18. Vance ED, Brookes PC, Jenkinson DS (1987) An extraction method for measuring soil microbial biomass C. Soil Biol Biochem 19:703–707

    Article  CAS  Google Scholar 

  19. Zak JC, Ziehr LL, Urbanczyk K, McHam JB, Yarborough K (1997) Big Bend National Park Watershed Program: monitoring microbial activity and diversity along an elevational gradient. Presented at the 9th Conference on Research and Management in Parks and Public Lands

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Acknowledgments

This work was funded by United States Geological Survey-BRD Small Watershed Program and National Park Service grants to JCZ. Access to sites was made possible by Joe Sirotnak (Big Bend National Park). Many helpful discussions were provided by Dora Ogles (Microbial Insights), Edward Sobek (Clean Air Labs), and Jennifer Humphries (Texas Tech University).

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Correspondence to James H. Campbell.

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Campbell, J.H., Clark, J.S. & Zak, J.C. PCR-DGGE Comparison of Bacterial Community Structure in Fresh and Archived Soils Sampled along a Chihuahuan Desert Elevational Gradient. Microb Ecol 57, 261–266 (2009). https://doi.org/10.1007/s00248-008-9479-3

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  • DOI: https://doi.org/10.1007/s00248-008-9479-3

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