Identification of marine algicidal Flavobacterium sp. 5 N-3 using multiple probes and whole-cell hybridization.

DOI Web Site 参考文献62件
  • ADACHI MASAO
    Laboratory of Aquatic Environmental Science, Faculty of Agriculture, Kochi University
  • FUKAMI KIMIO
    Laboratory of Aquatic Environmental Science, Faculty of Agriculture, Kochi University
  • KONDO RYUJI
    Department of Marine Bioscience, Fukui Prefectural University
  • NISHIJIMA TOSHITAKA
    Laboratory of Aquatic Environmental Science, Faculty of Agriculture, Kochi University

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We designed five 16S rRNA-targeted oligonucleotide probes (Sp probes) specific for Flavobacterium sp. 5 N-3, which inhibits the growth of red tide phytoplankton Gymnodinium mikimotoi (Dinophyceae). These probes were evaluated by whole-cell hybridization against 5 N-3 cells incubated under laboratory conditions. The fluorescence signal from the cell detected with Sp probe mix5, a mixture of the five probes, was 8.4-fold higher than that obtained with only one Sp probe (Sp01RF). The signal obtained by this method was strong enough to recognize 5 N-3 cells and count them under the epifluorescence microscope, while the signal was often undetectable when a single probe was used. Fluorescence intensities of cells at stationary phases and of ‘starved’ cells in sterile seawater using Sp probe mix5 were low but still sufficient for enumeration. These Sp probes did not hybridize with 11 strains from the Cytophaga/Flavobacteria/Bacteroides phylum and did react with strain 5 N-3 following whole-cell hybridization. These results show that 5 N-3 cells cultivated under our laboratory conditions can be detected by whole-cell hybridization with the five designed probes. These data also suggest that this technique may be useful for detection of an algicidal bacterium 5 N-3 in the natural environment.

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