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Development of specific oligonucleotide probes for the identification and in situ detection of hydrocarbon‐degrading Alcanivorax strains
Author(s) -
Syutsubo Kazuaki,
Kishira Hideo,
Harayama Shigeaki
Publication year - 2001
Publication title -
environmental microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.954
H-Index - 188
eISSN - 1462-2920
pISSN - 1462-2912
DOI - 10.1046/j.1462-2920.2001.00204.x
Subject(s) - biology , dapi , oligonucleotide , microbiology and biotechnology , 16s ribosomal rna , biostimulation , bacteria , dna , biochemistry , bioremediation , genetics , staining
The genus Alcanivorax comprises diverse hydrocarbon‐degrading marine bacteria. Novel 16S rRNA‐targeted oligonucleotide DNA probes (ALV735 and ALV735‐b) were developed to quantify two subgroups of the Alcanivorax / Fundibacter group by fluorescence in situ hybridization (FISH), and the conditions for the single‐mismatch discrimination of the probes were optimized. The specificity of the probes was improved further using a singly mismatched oligonucleotide as a competitor. The growth of Alcanivorax cells in crude oil‐contaminated sea water under the biostimulation condition was investigated by FISH with the probe ALV735, which targeted the main cluster of the Alcanivorax / Fundibacter group. The size of the Alcanivorax population increased with increasing incubation time and accounted for 91% of the 4′,6‐diamidino‐2‐phenylindole (DAPI) count after incubation for 2 weeks. The probes developed in this study are useful for detecting Alcanivorax populations in petroleum hydrocarbon‐degrading microbial consortia.