Unicellular diazotrophic cyanobacteira : development of immunocytochemical methods for nitrogenase detection and analysis of their role in Kuroshio Waters.

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Immunocytochemical methods that is applicable for detection of unicellular diazotrophic cyanobacteria in sea waters have been developed. Using developed methods, unicellular diazotrophic cyanobacteria were detected in sea waters collected from Kuroshio waters (off Ehime Prefecture, off Miyagi prefecture and off from South Taiwan). Detection of unicellular diazotrophic cyanobacteria was also tried using DNA-amplification of nitrogenase and 16SrRNA genes. Few diazotrophic unicellular cyanobacteria were detected by both immunocytochemical and DNA-based methods in waters from off Ehime and Miyagi. Unicellular diazotrophic cyanobacteria was abundant in waters from off Taiwan : cell densities were 0.5 to 1.5×103/L. Two diazotrophic cyanobacteria were successfully isolated from off Ehime (UWA1) and Taiwan (TW3). They were identified as Gloeothece spp. (UWA1) and Cyanothece spp. (TW3) from the partial sequences of 16S rRNA and nitrogenase (nifH) genes and their morphology.