title: Quantitative detection of selenate-reducing bacteria by real-time PCR targeting the selenate reductase gene.
abstract: We designed a primer set to target selenate reductase (SerA) for detecting selenate reducing bacteria (SeRB). Our serA gene-based PCR primer set has high specificity in that it and positively amplified some SeRB, but not denitrifying bacteria (DB). Phylogenetic analysis of serA clone sequences of environmental samples from selenate-reducing membrane biofilm reactor (MBfR) biofilms showed that these sequences were closely grouped and had high similarity to selenate reductase gene sequences from SeRB Thauera selenatis and DB Dechloromonas; however, they were distant to other genes from dimethylsulfoxide (DMSO) enzyme family. Constructing a standard curve targeting the serA gene, we found that the good linearity for the qPCR assay when applied it to quantify SeRB in MBfR biofilms, and the gene copies of SeRB correlated well to the selenate removal percentages. Our results demonstrated the feasibility of using the serA gene-based PCR primer set to detect and quantify SeRB in environmental samples.
authors.affiliation: Zhejiang Prov Key Lab Water Pollut Control & Envi, Zhejiang University, Hangzhou, Zhejiang, China; MOE Key Lab of Environmental Remediation and Ecosystem Health, Zhejiang University, Hangzhou, China; Department of Environmental Engineering, College of Environmental and Resource Science, Zhejiang University, Hangzhou, China.
authors.forename: Li-Lian
authors.initials: LL
authors.lastname: Wen
authors.affiliation: Zhejiang Prov Key Lab Water Pollut Control & Envi, Zhejiang University, Hangzhou, Zhejiang, China.
authors.forename: Chun-Yu
authors.initials: CY
authors.lastname: Lai
authors.affiliation: Hangzhou Institute of Environmental Protection Science, Hangzhou, China.
authors.forename: Qiang
authors.initials: Q
authors.lastname: Yang
authors.affiliation: Zhejiang Prov Key Lab Water Pollut Control & Envi, Zhejiang University, Hangzhou, Zhejiang, China.
authors.forename: Jia-Xian
authors.initials: JX
authors.lastname: Chen
authors.affiliation: Zhejiang Prov Key Lab Water Pollut Control & Envi, Zhejiang University, Hangzhou, Zhejiang, China.
authors.forename: Yin
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Swette Center for Environmental Biotechnology, Biodesign Institute at Arizona State University, P.O. Box 875701, Tempe, AZ 85287-5701, United States.
authors.forename: Aura
authors.initials: A
authors.lastname: Ontiveros-Valencia
authors.affiliation: Zhejiang Prov Key Lab Water Pollut Control & Envi, Zhejiang University, Hangzhou, Zhejiang, China; MOE Key Lab of Environmental Remediation and Ecosystem Health, Zhejiang University, Hangzhou, China; Department of Environmental Engineering, College of Environmental and Resource Science, Zhejiang University, Hangzhou, China. Electronic address: zhaohp@zju.edu.cn.
authors.forename: He-Ping
authors.initials: HP
authors.lastname: Zhao
mesh_terms: D001419:Bacteria / Q000201:enzymology* / Q000235:genetics*; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism*; D018441:Biofilms; D005798:Genes, Bacterial; D010084:Oxidation-Reduction; D010088:Oxidoreductases / Q000235:genetics* / Q000378:metabolism*; D010802:Phylogeny; D060888:Real-Time Polymerase Chain Reaction; D064586:Selenic Acid / Q000378:metabolism*; D016415:Sequence Alignment
chemical_list: D001426:Bacterial Proteins; D010088:Oxidoreductases; C078108:selenate reductase; D064586:Selenic Acid
keywords: DMSO; Membrane biofilm reactor; Primer design; Selenate reductase