title: Reassessment of PCR primers targeting 16S rRNA genes of the organohalide-respiring genus Dehalogenimonas.
abstract: Representatives from the genus Dehalogenimonas have the metabolic capacity to anaerobically transform a variety of environmentally important polychlorinated aliphatic compounds. In light of the recent isolation of additional strains, description of a new species, and an expanded number of uncultured DNA sequences, PCR primers and protocols intended to uniquely target members of this organohalide-respiring genus were reevaluated. Nine of fourteen primer combinations reported previously as genus-specific failed to amplify 16S rRNA genes of recently isolated Dehalogenimonas strains. Use of alternative combinations or modified genus-specific primers, however, allowed detection of all presently known Dehalogenimonas strains. Use of a modified primer set in qPCR revealed an approximately two-order of magnitude increase in concentration of Dehalogenimonas 16S rRNA gene copies following subsurface injection of electron donors at a Louisiana Superfund site, demonstrating the utility of the newly developed protocol and suggesting that the genus Dehalogenimonas can respond to biostimulation remediation strategies in a manner similar to that previously reported for other dechlorinating genera such as Dehalococcoides.
authors.affiliation: Department of Civil and Environmental Engineering, Louisiana State University, 3513B Patrick Taylor Hall, Baton Rouge, LA, 70803, USA.
authors.forename: Jie
authors.initials: J
authors.lastname: Chen
authors.forename: Kimberly S
authors.initials: KS
authors.lastname: Bowman
authors.forename: Fred A
authors.initials: FA
authors.lastname: Rainey
authors.forename: William M
authors.initials: WM
authors.lastname: Moe
mesh_terms: D001673:Biodegradation, Environmental; D041862:Chloroflexi / Q000378:metabolism*; D017931:DNA Primers / Q000235:genetics; D016133:Polymerase Chain Reaction; D012336:RNA, Ribosomal, 16S / Q000235:genetics*
chemical_list: D017931:DNA Primers; D012336:RNA, Ribosomal, 16S