Microbial Ecology

, Volume 69, Issue 1, pp 59–65 | Cite as

Isolation, Characterization and Community Diversity of Indigenous Putative Toluene-Degrading Bacterial Populations with Catechol-2,3-Dioxygenase Genes in Contaminated Soils

  • Ola A. Olapade
  • Adam J. Ronk
Environmental Microbiology


Indigenous bacterial assemblages with putative hydrocarbon-degrading capabilities were isolated, characterized and screened for the presence of the catechol-2,3-dioxygenase (C23O) gene after exposure to toluene in two different (i.e., pristine and conditioned) soil communities. The indigenous bacterial populations were exposed to the hydrocarbon substrate by the addition of toluene concentrations, ranging from 0.5 % to 10 % V/W in 10 g of each soil and incubated at 30 °C for upwards of 12 days. In total, 25 isolates (11 in pristine soil and 14 in conditioned soil) were phenotypically characterized according to standard microbiological methods and also screened for the 238-bp C23O gene fragment. Additionally, 16S rRNA analysis of the isolates identified some of them as belonging to the genera Bacillus, Exiguobacterium, Enterobacter, Pseudomonas and Stenotrophomonas. Furthermore, the two clone libraries that were constructed from these toluene-contaminated soils also revealed somewhat disparate phylotypes (i.e., 70 % Actinobacteria and Firmicutes to 30 % Proteobacteria in conditioned soil, whereas in pristine soil: 66 % Actinobacteria and Firmicutes; 21 % Proteobacteria and 13 % Bacteroidetes). The differences observed in bacterial phylotypes between these two soil communities may probably be associated with previous exposure to hydrocarbon sources by indigenous populations in the conditioned soil as compared to the pristine soil.


Clone Library Proteobacteria Actinobacteria Firmicutes Bacteroidetes 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.



The study was supported mostly by the Albion College Hewlett-Mellon Faculty Development Funds to OAO for the 2013 summer session. We particularly thank the staff members of Albion Motors for granting unrestricted access to their premises during soil sampling. Also our sincere appreciation goes to Melissa Goodell, Kurt Hellman, Dave Carey, Freyja Davis and Lori Duff for assistance with various supplies and materials that were needed during the study period.


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Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  1. 1.Department of Biology and the Center for Sustainability and the EnvironmentAlbion CollegeAlbionUSA

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