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Applied and Environmental Microbiology, September 2006, p. 6161-6168, Vol. 72, No. 9
0099-2240/06/$08.00+0     doi:10.1128/AEM.01070-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Reductive Dehalogenase Gene Expression as a Biomarker for Physiological Activity of Dehalococcoides spp.

Patrick K. H. Lee,1 David R. Johnson,1 Victor F. Holmes,1 Jianzhong He,1,{dagger} and Lisa Alvarez-Cohen1,2*

Department of Civil and Environmental Engineering, University of California, Berkeley, California 94720-1170,1 Earth Sciences Division, Lawrence Berkeley National Laboratory, 1 Cyclotron Rd., Berkeley, California 947202

Received 9 May 2006/ Accepted 29 June 2006

This study characterizes the transcriptional expression of the reductive dehalogenase (RDase)-encoding tceA and vcrA genes and evaluates their applicability as potential biological markers of Dehalococcoides activity. When Dehalococcoides ethenogenes 195 was provided with trichloroethene (TCE) as the electron acceptor, the expression of the tceA gene increased by 90-fold relative to that in cells starved of chlorinated ethenes, demonstrating that tceA gene expression is indicative of the active physiological state of this strain. In a Dehalococcoides-containing enrichment culture that contains both the tceA and vcrA genes, the tceA gene was up-regulated in response to TCE and cis-1,2-dichloroethene (cDCE) exposure, while the vcrA gene was up-regulated in response to TCE, cDCE, and vinyl chloride (VC). When chlorinated ethenes were depleted, the RDase-encoding gene transcripts decayed exponentially, with a half-life between 4.8 and 6.1 h, until they reached a stable background level after 2 days. We found that while gene expression correlated generally to the presence of chlorinated ethenes, there was no apparent direct relationship between RDase-encoding transcript numbers and respective rates of TCE, cDCE, and VC dechlorination activities. However, elevated tceA and vcrA expression did correlate with chlorinated-ethene reduction beyond cDCE, suggesting that elevated RDase-encoding transcript numbers could serve as a biomarker for the physiological ability of Dehalococcoides spp. to dechlorinate beyond cDCE.

* Corresponding author. Mailing address: 726 Davis Hall, Department of Civil and Environmental Engineering, University of California, Berkeley, CA 94720-1710. Phone: (510) 643-5969. Fax: (510) 642-7483. E-mail: alvarez{at}ce.berkeley.edu.

{dagger} Present address: Division of Environmental Science and Engineering, National University of Singapore, Singapore 117576, Republic of Singapore.

Applied and Environmental Microbiology, September 2006, p. 6161-6168, Vol. 72, No. 9
0099-2240/06/$08.00+0     doi:10.1128/AEM.01070-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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