Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Monitoring Bacterial Population Dynamics Using Real-Time PCR During the Bioremediation of Crude-Oil-Contaminated Soil

  • Baek, Kyung-Hwa (Environmental Biotechnology Research Center, KRIBB) ;
  • Yoon, Byung-Dae (Environmental Biotechnology Research Center, KRIBB) ;
  • Cho, Dae-Hyun (Environmental Biotechnology Research Center, KRIBB) ;
  • Kim, Byung-Hyuk (Environmental Biotechnology Research Center, KRIBB) ;
  • Oh, Hee-Mock (Environmental Biotechnology Research Center, KRIBB) ;
  • Kim, Hee-Sik (Environmental Biotechnology Research Center, KRIBB)
  • Published : 2009.04.30
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Abstract

We evaluated the activity and abundance of the crude-oil-degrading bacterium Nocardia sp. H17-1 during bioremediation of oil-contaminated soil, using real-time PCR. The total petroleum hydrocarbon(TPH) degradation rate constants(k) of the soils treated with and without H17-1 were $0.103\;d^{-1}$ and $0.028\;d^{-1}$ respectively. The degradation rate constant was 3.6 times higher in the soil with H17-1 than in the soil without H17-1. In order to detect and quantify the Nocardia sp. H17-1 in soil samples, we quantified the genes encoding 16S ribosomal RNA(16S rRNA), alkane monooxygenase(alkB4), and catechol 2,3-dioxygenase(23CAT) with real-time PCR using SYBR green. The amounts of H17-1 16S rRNA and alkB4 detected increased rapidly up to 1,000-folds for the first 10 days, and then continued to increase only slightly or leveled off. However, the abundance of the 23CAT gene detected in H17-1-treated soil, where H17-1 had neither the 23CAT gene for the degradation of aromatic hydrocarbons nor the catechol 2,3-dioxygenase activity, did not differ significantly from that of the untreated soil($\alpha$=0.05,p>0.22). These results indicated that H17-1 is a potential candidate for the bioaugmentation of alkane-contaminated soil. Overall, we evaluated the abundance and metabolic activity of the bioremediation strain H17-1 using real-time PCR, independent of cultivation.

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References

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