title: Fluorescence-based bacterial bioreporter for specific detection of methyl halide emissions in the environment.
abstract: Methyl halides are volatile one-carbon compounds responsible for substantial depletion of stratospheric ozone. Among them, chloromethane (CH3Cl) is the most abundant halogenated hydrocarbon in the atmosphere. Global budgets of methyl halides in the environment are still poorly understood due to uncertainties in their natural sources, mainly from vegetation, and their sinks, which include chloromethane-degrading bacteria. A bacterial bioreporter for the detection of methyl halides was developed on the basis of detailed knowledge of the physiology and genetics of Methylobacterium extorquens CM4, an aerobic alphaproteobacterium which utilizes chloromethane as the sole source of carbon and energy. A plasmid construct with the promoter region of the chloromethane dehalogenase gene cmuA fused to a promotorless yellow fluorescent protein gene cassette resulted in specific methyl halide-dependent fluorescence when introduced into M. extorquens CM4. The bacterial whole-cell bioreporter allowed detection of methyl halides at femtomolar levels and quantification at concentrations above 10 pM (approximately 240 ppt). As shown for the model chloromethane-producing plant Arabidopsis thaliana in particular, the bioreporter may provide an attractive alternative to analytical chemical methods to screen for natural sources of methyl halide emissions.
authors: Université de Strasbourg, Equipe Adaptations et Interactions Microbiennes dans l'Environnement, Département Microorganismes, Génomes, Environnement, UMR 7156 UdS-CNRS Génétique Moléculaire, Génomique, Microbiologie, Strasbourg, France.
authors: Muhammad
authors: M
authors: Farhan Ul Haque
authors: Thierry
authors: T
authors: Nadalig
authors: Françoise
authors: F
authors: Bringel
authors: Hubert
authors: H
authors: Schaller
authors: Stéphane
authors: S
authors: Vuilleumier
mesh_terms: D017360:Arabidopsis / Q000378:metabolism; D001426:Bacterial Proteins / Q000235:genetics; D017931:DNA Primers / Q000235:genetics; D004784:Environmental Monitoring / Q000379:methods*; D004785:Environmental Pollutants / Q000032:analysis*; D005470:Fluorometry; D008164:Luminescent Proteins / Q000235:genetics; D008737:Methyl Chloride / Q000032:analysis* / Q000378:metabolism; D020601:Methylobacterium extorquens / Q000201:enzymology*; D008780:Methyltransferases / Q000235:genetics / Q000378:metabolism*; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D010957:Plasmids / Q000235:genetics; D011401:Promoter Regions, Genetic / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D001426:Bacterial Proteins; D017931:DNA Primers; D004785:Environmental Pollutants; D008164:Luminescent Proteins; C054562:yellow fluorescent protein, Bacteria; D008737:Methyl Chloride; D008780:Methyltransferases
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