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Applied and Environmental Microbiology, December 2005, p. 8257-8264, Vol. 71, No. 12
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.12.8257-8264.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Multiple Reductive-Dehalogenase-Homologous Genes Are Simultaneously Transcribed during Dechlorination by Dehalococcoides-Containing Cultures

Alison S. Waller,1 Rosa Krajmalnik-Brown,2 Frank E. Löffler,2,3 and Elizabeth A. Edwards1*

Department of Chemical Engineering and Applied Chemistry, University of Toronto, Toronto, Ontario, Canada,1 School of Civil and Environmental Engineering, Georgia Institute of Technology, Atlanta, Georgia,2 School of Biology, Georgia Institute of Technology, Atlanta, Georgia3

Received 10 June 2005/ Accepted 3 September 2005

Degenerate primers were used to amplify 14 distinct reductive-dehalogenase-homologous (RDH) genes from the Dehalococcoides-containing mixed culture KB1. Most of the corresponding predicted proteins were highly similar (97 to >99% amino acid identity) to previously reported Dehalococcoides reductive dehalogenases. To examine the differential transcription of these RDH genes, KB1 was split into five subcultures amended with either trichloroethene, cis-1,2-dichloroethene, vinyl chloride, 1,2-dichlorethane, or no chlorinated electron acceptor. Total RNA was extracted following the onset of reductive dechlorination, and RDH transcripts were reverse transcribed and amplified using degenerate primers. The results indicate that the transcription of RDH genes requires the presence of a chlorinated electron acceptor, and for all treatments, multiple RDH genes were simultaneously transcribed, with transcripts of two of the genes being present under all four electron-accepting conditions. Two of the transcribed sequences were highly similar to reported vinyl chloride reductase genes, namely, vcrA from Dehalococcoides sp. strain VS and bvcA from Dehalococcoides sp. strain BAV1. These findings suggest that multiple RDH genes are induced by a single chlorinated substrate and that multiple reductive dehalogenases contribute to chloroethene degradation in KB1.

* Corresponding author. Mailing address: Department of Chemical Engineering and Applied Chemistry, University of Toronto, 200 College St., Toronto, Ontario M5S 3E5, Canada. Phone: (416) 946-3506. Fax: (416) 978-8605. E-mail: edwards{at}chem-eng.utoronto.ca.

Applied and Environmental Microbiology, December 2005, p. 8257-8264, Vol. 71, No. 12
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.12.8257-8264.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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