In situ detection of alkB2 gene involved in Alcanivorax borkumensis SK2(T) hydrocarbon biodegradation.
This study aimed to develop a new assay based on the whole cell hybridization in order to monitor alkane hydroxylase genes (alkB system) of the marine bacterium Alcanivorax borkumensis SK2(T) commonly reported as the predominant microorganism responsible for the biodegradation of n-alkanes which are the major fraction of petroleum hydrocarbons. The assay based on the whole cell hybridization targeting alkB2 gene was successfully developed and calibrated on a pure culture of Alcanivorax borkumensis SK2(T) with a detection efficiency up to 80%. The approach was further successfully validated on hydrocarbon-contaminated seawater and provided cells abundance (6.74E+04alkB2-carryingcellsmL(-1)) higher of about one order of magnitude than those obtained by qPCR (4.96E+03alkB2genecopiesmL(-1)). This study highlights the validity of the assay for the detection at single cell level of key-functional genes involved in the biodegradation of n-alkanes.
Water Research Institute, IRSA-CNR, Via Salaria km 29,300, Monterotondo, RM, Italy.
Bruna
B
Matturro
Water Research Institute, IRSA-CNR, Via Salaria km 29,300, Monterotondo, RM, Italy.
Emanuela
E
Frascadore
Institute of Marine and Coastal Environments, IAMC-CNR, Spianata S. Raineri, 86, Messina, ME, Italy.
Simone
S
Cappello
Institute of Marine and Coastal Environments, IAMC-CNR, Spianata S. Raineri, 86, Messina, ME, Italy.
Mariella
M
Genovese
Water Research Institute, IRSA-CNR, Via Salaria km 29,300, Monterotondo, RM, Italy. Electronic address: rossetti@irsa.cnr.it.
Simona
S
Rossetti
D053521:Alcanivoraceae / Q000235:genetics* / Q000378:metabolism*; D000473:Alkanes / Q000378:metabolism; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D001673:Biodegradation, Environmental; D042926:Cytochrome P-450 CYP4A / Q000235:genetics* / Q000378:metabolism; D006838:Hydrocarbons / Q000378:metabolism*; D017404:In Situ Hybridization, Fluorescence; D007558:Italy; D010578:Petroleum / Q000378:metabolism; D015203:Reproducibility of Results; D012623:Seawater / Q000382:microbiology*; D014874:Water Pollutants, Chemical / Q000378:metabolism
D000473:Alkanes; D001426:Bacterial Proteins; D006838:Hydrocarbons; D010578:Petroleum; D014874:Water Pollutants, Chemical; D042926:Cytochrome P-450 CYP4A
Alcanivorax borkumensis SK2(T); GeneCARD-FISH; Petroleum-polluted seawater; alkB2 gene; n-Alkanes