The marine planktonic cyanobacterial genus Katagnymene was described by Lemmermann in 1900 and is found in oligotrophic tropical and subtropical oceans. The genus comprises two species, K. pelagica Lemmermann and K. spiralis Lemmermann, and both were observed in most stations sampled during a cruise in the southwest Pacific Ocean in 1998. Katagnymene is nonheterocystous and characterized by single ensheathed trichomes that do not form colonies. Acetylene reduction-GC demonstrated that natural populations of Katagnymene fixed nitrogen and that nitrogenase activity occurred exclusively in the light during a 12:12 light:dark cycle. Whole cell immunolocalization revealed that nitrogenase (the Fe protein) appeared in 7% of the total number of cells and that these were arranged in zones composed of consecutively arranged cells. At least one zone of nitrogenase-containing cells per trichome was found. Nitrogenase was present throughout both the day and night, as shown by Western blotting, and in the same percentage of cells. Ultrastructural immunolocalization on sectioned trichomes also confirmed the presence and localization of nitrogenase in Katagnymene. Cultures of Katagnymene are able to grow on nitrogen-free media and fix nitrogen only in the light. Finally, cloning and sequencing of nifH verified the diazotrophic nature of the genus Katagnymene and demonstrated a close relationship to members of the marine genus Trichodesmium.