Evaluation of soil-microbial communities by their CLPP. Standardization of a laboratory technique to replace commercial available microplates

Authors

  • Luciana P. Di Salvo Cátedra de Microbiología Agrícola, Facultad de Agronomía, Universidad de Buenos Aires, Ciudad de Buenos Aires, Argentina
  • Inés E. García de Salamone Cátedra de Microbiología Agrícola, Facultad de Agronomía, Universidad de Buenos Aires, Ciudad de Buenos Aires, Argentina

Keywords:

carbon-source utilization, community-level physiological profiles, functional diversity, metabolic profiles

Abstract

Variation of soil-microbial communities are good bioindicators of human impacts in soils, such as different soils management or contamination. Considering that traditional methods of isolation and taxonomic analysis do not consider the functionality of the microbial community, Community-Level Physiological Profi les (CLPP) emerged as a complementary methodology to study microbial communities. Several studies have shown that Biolog® EcoPlates® are very useful for determining physiological differences between communities from different samples. However, commercial microplates have some disadvantages which led us to the idea of replacing them by microplates prepared in the laboratory (Laboratory’s). Here, we compared both types of microplates using soil samples from a bioremediation assay. We compared a) the average well color development for each sample, b) the averages of absorbance values for each type of microplate, c) Principal Components, and d) Shannon-Weaver’s diversity index (H). Although Laboratory’s showed signifi cantly lower Average absorbance values than EcoPlates®, the principal component analysis and diversity index did not differ between types of microplates. In conclusion, both types of microplates showed a relatively similar ability to detect differences in the CLPP of the treatments studied. Consequently, microplates prepared in laboratory are a reliable and economical tool to study the physiology of soil microbial communities.

References

ABRIL, A. 2003. ¿Son los microorganismos edáficos buenos indicadores de impacto productivo en los ecosistemas? Ecología Austral 13:195-204.

BAKKEN, LR. 1997. Culturable and nonculturable bacteria in soil. In: van Elsas, JD; JT Trevors & EMH Wellington (eds.). Modern Soil Microbiology. Marcel Dekker.

BAUDOIN, E; E BENIZRI & A GUCKERT. 2002. Impact of growth stage on the bacterial community structure along maize roots, as determined by metabolic and genetic fingerprinting. Appl. Soil Ecol. 19:135-145.

BAUDOIN, E; E BENIZRI & A GUCKERT. 2003. Impact of artificial root exudates on the bacterial community structure in bulk soil and maize rhizosphere. Soil Biol. and Biochem. 35:1183-1192.

BEARE, MH; RW PARMELEE; PF HENDRIX; W CHENG; DC COLEMAN; ET AL. 1992. Microbial and faunal interactions and effects on litter nitrogen and descomposition in agroecosystems. Ecological Monographs 62:569-591.

BENDING, GD; MK TURNER; F RAYNS; MC MARX & M WOOD. 2004. Microbial and biochemical soil quality indicators and their potential for differentiating areas under contrasting agricultural management regimes. Soil Biol. and Biochem. 36:1785-1792.

BOCHNER, BR & MA SAVAGEAU. 1977. Generalized indicator plate for genetic, metabolic, and taxonomic studies with microorganisms. Appl. Environ. Microbiol. 33:434-444.

BOSSIO, DA & KM SCOW. 1995. Impact of carbon and flooding on the metabolic diversity of microbial communities in soils. Appl. Environ. Microbiol. 61:4043-4050.

BUCHER, AE & LE LANYON. 2005. Evaluating soil management with microbial community-level physiological profi les. Appl. Soil Ecol. 29:59-71.

CALBRIX, R; K LAVAL & S BARRAY. 2005. Analysis of the potential functional diversity of the bacterial community in soil: a reproducible procedure using sole-carbon-source utilization profiles. European J. of Soil Biol. 41:11-20.

CAMPBELL, CD; SJ GRAYSTON & DJ HIRST. 1997. Use of rhizosphere carbon sources in sole carbon source tests to discriminate soil microbial communities. J. of Microbiol. Methods 30:33-41.

DI SALVO, LP; J ESCOBAR ORTEGA; S TORRI; MS ZUBILLAGA & IE GARCÍA DE SALAMONE. 2007. Influencia del raigrás y la fertilización sobre las comunidades microbianas de un suelo contaminado con fenantreno. Actas XI Congreso Argentino y Latinoamericano de Microbiología. Córdoba, Argentina.

DERRY, AM; WJ STADDON & JT TREVORS. 1998. Functional diversity and community structure of microorganisms in uncontaminated and creosote-contaminated soils as determined by sole-carbon-source-utilization. World J. of Microbiol. and Biotechnology 14:571-578.

DOBLER, R; P BURRI; K GRUIZ; H BRANDL & R BACHOFEN. 2001. Variability in microbial populations in soil higly polluted with heavy metals on the basis of substrate utilization pattern analysis. J. Soils and Sediments 1(3):151-158.

DOBLER, R; M SANER & R BACHOFEN. 2000. Population changes of soil microbial communties indiced by hydrocarbon and heavy metal contamination. Bioremediaton J. 4:41-56.

ELLIS, RJ; P MORGAN; AJ WEIGHTMAN & JC FRY. 2003. Cultivation-dependent and independent approaches for determining bacterial diversity in heavy-metal-contaminated soil. Appl. and Environ. Microb. 69(6):3223-3230.

ENGELEN, B; K MEINKEN; F VON WINTZINGERODE; H HEUER; HP MALKOMES; ET AL. 1998. Monitoring impact of a pesticide treatment on bacterial soil communities by metabolic and genetic fi ngerprinting in addition to conventional testing procedures. Appl. Environ. Microbiol. 64:2814-2821.

GARCÍA DE SALAMONE, IE; LP DI SALVO; JS ESCOBAR ORTEGA; PMF BOA SORTE; S URQUIAGA; ET AL. 2010. Field response of rice paddy crop to Azospirillum inoculation: physiology of rhizosphere bacterial communities and the genetic diversity of endophytic bacteria in different parts of the plants. Plant and Soil 336:351-362.

GARLAND, JL. 1996. Analytical approaches to the characterization of samples of microbial communities using patterns of potential C source utilization. Soil. Biol. and Biochem. 28(2):213-221.

GARLAND, JL & AL MILLS. 1991. Classification and characterization of heterotrophic microbial communities on the basis of patterns of community-level sole-carbon-source utilization. Appl. and Environ. Microbiol. 57(8):2351-2359.

GÓMEZ, E; J GARLAND & M CONTI. 2004. Reproducibility in the response of soil bacterial community-level physiological profi les from a land use intensifi cation gradient. Appl. Soil Ecol. 26:21-30.

HAACK, SK; H GARCHOW; MJ KLUG & L FORNEY. 1995. Analysis of factors affecting the accuracy, reproducibility, and interpretation of microbial community carbon source utilization patterns. Appl. Environ. Microbiol. 61(4):1458-1468.

INSAM, H. 1997. A new set of substrates proposed for community characterization in environmental samples. In: Insam, H & A Rangger (eds.). Microbial communities: Functional versus structural approaches. Springer, Berlin.

KELLY, JJ & RLI TATE. 1998. Use of Biolog for the analysis of microbial communities form zinc-contaminated soils. J. Environ. Qual. 27:600-608.

LAWLOR, K; BP KNIGHT; VL BAROSA-JEFFERSON; PW LANE; AK LILLEY; ET AL. 2000. Comparison of methods to investigate microbial populations in soils under different agricultural management. FEMS Microbiol. Ecol. 33:129-137.

MOYNAHAN, OS; CA ZABINSKI & JE GANNON. 2002. Microbial community structure and carbon-utilization diversity in a mine tailings revegetation study. Restoration Ecology 10:77-87.

NAIMAN, AD; A LATRÓNICO & IE GARCÍA DE SALAMONE. 2009. Inoculation of wheat with Azospirillum brasilense and Pseudomonas fluorescens: Impact on the roduction and culturable rhizosphere microflora. European J. of Soil Biol. 45:44-51.

NIELSEN, MN & AWINDING. 2002. Microorganisms as indicators of soil health. National Environmental Research Institute, Denmark. Technical Report No. 388.

PALMROTH, MRT; U MÜNSTER; J PICHTEL & JA PUHAKKA. 2005. Metabolic responses of microbiota to diesel fuel addition in vegetated soil. Biodegradation 16:91-101.

PRESTON-MAFHAM, J; L BODDY & PF RANDERSON. 2002. Analysis of microbial community functional diverstiy using sole-carbon-source utilisation profi les - a critique. FEMS Microbiol. Ecol. 42:1-14.

SCHNÜRER, J; M. CLARHOLM & ROSSWALL T. 1985. Microbial biomass and activity in an agricultural soil with different organic matter contents. Soil Biol. and Biochem. 17:611-618.

SEMMARTIN, M; C DI BELLA & IE GARCÍA DE SALAMONE. 2010. Grazing-induced changes in plant species composition affect plant and soil properties of grassland mesocosms. Plant and Soil 328:471-481.

ZAK, HC; MR WILLIG; DL MOORHEAD & HG WILDMAN. 1994. Functional diversity of microbial communities: a quantitative approach. Soil Biol. and Biochem. 26:1101-1108.

Downloads

Published

2012-08-01

How to Cite

Di Salvo, L. P., & García de Salamone, I. E. (2012). Evaluation of soil-microbial communities by their CLPP. Standardization of a laboratory technique to replace commercial available microplates. Ecología Austral, 22(2), 129–136. Retrieved from https://ojs.ecologiaaustral.com.ar/index.php/Ecologia_Austral/article/view/1256

Issue

Section

Short Communications