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KMID : 0545120090190040339
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Journal of Microbiology and Biotechnology
2009 Volume.19 No. 4 p.339 ~ p.345
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Monitoring Bacterial Population Dynamics Using Real-Time PCR During the Bioremediation of Crude-Oil-Contaminated Soil
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Baek Kyung-Hwa
Yoon Byung-Dae
Cho Dae-Hyun
Kim Byung-Hyuk
Oh Hee-Mock
Kim Hee-Sik
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Abstract
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We evaluated the activity and abundance of the crudeoil- 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 (α=0.05, p>0.22).
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KEYWORD
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Bioaugmentation, crude oil, Nocardia sp., real-time PCR, total petroleum hydrocarbon
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