Isolation of isoprene degrading bacteria from soils, development of isoA gene probes and identification of the active isoprene-degrading soil community using DNA-stable isotope probing.
Emissions of biogenic volatile organic compounds (bVOCs), are an important element in the global carbon cycle, accounting for a significant proportion of fixed carbon. They contribute directly and indirectly to global warming and climate change and have a major effect on atmospheric chemistry. Plants emit isoprene to the atmosphere in similar quantities to emissions of methane from all sources and each accounts for approximately one third of total VOCs. Although methanotrophs, capable of growth on methane, have been intensively studied, we know little of isoprene biodegradation. Here, we report the isolation of two isoprene-degrading strains from the terrestrial environment and describe the design and testing of polymerase chain reaction (PCR) primers targeting isoA, the gene encoding the active-site component of the conserved isoprene monooxygenase, which are capable of retrieving isoA sequences from isoprene-enriched environmental samples. Stable isotope probing experiments, using biosynthesized (13) C-labelled isoprene, identified the active isoprene-degrading bacteria in soil. This study identifies novel isoprene-degrading strains using both culture-dependent and, for the first time, culture-independent methods and provides the tools and foundations for continued investigation of the biogeography and molecular ecology of isoprene-degrading bacteria.
University of East Anglia, Norwich Research Park, Norwich, UK.
Myriam
M
El Khawand
University of East Anglia, Norwich Research Park, Norwich, UK.
Andrew T
AT
Crombie
University of East Anglia, Norwich Research Park, Norwich, UK.
Antonia
A
Johnston
DuPont Industrial Biosciences, 925 Page Mill Road, Palo Alto, CA, 94304, USA.
Dmitrii V
DV
Vavlline
DuPont Industrial Biosciences, 925 Page Mill Road, Palo Alto, CA, 94304, USA.
Joseph C
JC
McAuliffe
DuPont Industrial Biosciences, 925 Page Mill Road, Palo Alto, CA, 94304, USA.
Jacob A
JA
Latone
DuPont Industrial Biosciences, 925 Page Mill Road, Palo Alto, CA, 94304, USA.
Yuliya A
YA
Primak
DuPont Industrial Biosciences, 925 Page Mill Road, Palo Alto, CA, 94304, USA.
Sang-Kyu
SK
Lee
DuPont Industrial Biosciences, 925 Page Mill Road, Palo Alto, CA, 94304, USA.
Gregg M
GM
Whited
University of Essex, Wivenhoe Park, Colchester, UK.
Terry J
TJ
McGenity
University of East Anglia, Norwich Research Park, Norwich, UK.
J Colin
JC
Murrell
D001272:Atmosphere / Q000032:analysis; D001419:Bacteria / Q000235:genetics / Q000378:metabolism*; D001483:Base Sequence; D002070:Butadienes / Q000378:metabolism*; D057486:Carbon Cycle / Q000502:physiology*; D057231:Climate Change; D004247:DNA / Q000378:metabolism; D015342:DNA Probes; D004269:DNA, Bacterial / Q000235:genetics; D016680:Genome, Bacterial / Q000235:genetics; D045782:Hemiterpenes / Q000378:metabolism*; D008697:Methane / Q000378:metabolism; D010105:Oxygenases / Q000378:metabolism*; D010420:Pentanes / Q000378:metabolism*; D010944:Plants / Q000378:metabolism*; D017422:Sequence Analysis, DNA; D012987:Soil; D012988:Soil Microbiology; D055549:Volatile Organic Compounds / Q000378:metabolism*
D002070:Butadienes; D015342:DNA Probes; D004269:DNA, Bacterial; D045782:Hemiterpenes; D010420:Pentanes; D012987:Soil; D055549:Volatile Organic Compounds; C005059:isoprene; D004247:DNA; D010105:Oxygenases; D008697:Methane