Quantitative PCR confirms purity of strain GT, a novel trichloroethene-to-ethene-respiring Dehalococcoides isolate

Applied and Environmental Microbiology
Youlboong SungFrank E Löffler

Abstract

A novel Dehalococcoides isolate capable of metabolic trichloroethene (TCE)-to-ethene reductive dechlorination was obtained from contaminated aquifer material. Growth studies and 16S rRNA gene-targeted analyses suggested culture purity; however, the careful quantitative analysis of Dehalococcoides 16S rRNA gene and chloroethene reductive dehalogenase gene (i.e., vcrA, tceA, and bvcA) copy numbers revealed that the culture consisted of multiple, distinct Dehalococcoides organisms. Subsequent transfers, along with quantitative PCR monitoring, yielded isolate GT, possessing only vcrA. These findings suggest that commonly used qualitative 16S rRNA gene-based procedures are insufficient to verify purity of Dehalococcoides cultures. Phylogenetic analysis revealed that strain GT is affiliated with the Pinellas group of the Dehalococcoides cluster and shares 100% 16S rRNA gene sequence identity with two other Dehalococcoides isolates, strain FL2 and strain CBDB1. The new isolate is distinct, as it respires the priority pollutants TCE, cis-1,2-dichloroethene (cis-DCE), 1,1-dichloroethene (1,1-DCE), and vinyl chloride (VC), thereby producing innocuous ethene and inorganic chloride. Strain GT dechlorinated TCE, cis-DCE, 1,1-DCE, and VC to ...Continue Reading

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Citations

Sep 5, 2007·Applied Microbiology and Biotechnology·Michael BungeJosef Zeyer
Jul 21, 2009·Applied Microbiology and Biotechnology·Michael Bunge, Ute Lechner
Jun 13, 2009·Journal of Industrial Microbiology & Biotechnology·Simon VainbergRobert J Steffan
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Nov 19, 2011·Environmental Science & Technology·Patrick K H LeeLisa Alvarez-Cohen
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Jul 17, 2009·Environmental Science & Technology·Concepción Carreón-DiazcontiMark L Brusseau
Mar 13, 2013·Philosophical Transactions of the Royal Society of London. Series B, Biological Sciences·Jun YanFrank E Löffler
Mar 13, 2013·Philosophical Transactions of the Royal Society of London. Series B, Biological Sciences·Christian J SchippLorenz Adrian
Mar 13, 2013·Philosophical Transactions of the Royal Society of London. Series B, Biological Sciences·Laura A HugLorenz Adrian
Jun 15, 2011·Applied and Environmental Microbiology·Laura A HugElizabeth A Edwards
Sep 8, 2006·Applied and Environmental Microbiology·Patrick K H LeeLisa Alvarez-Cohen
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Dec 20, 2011·Applied and Environmental Microbiology·Patrick K H LeeLisa Alvarez-Cohen
Apr 7, 2006·Applied and Environmental Microbiology·Kirsti M RitalahtiFrank E Löffler

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