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We examined diazotroph activity in controlled experiments containing natural populations of nitrogen (N2)- fixing microbes collected offshore from Kane'ohe Bay, Hawai'i, and from Station ALOHA (A Long-term Oligotrophic Habitat Assessment) in the North Pacific subtropical gyre. Quantitative polymerase chain reaction and reverse transcription-polymerase chain reaction approaches showed that the abundances of the major phylotypes did not change substantially during experimental incubations. Diel phasing of nifH gene expression in the experiments was similar to that documented for natural populations. Cell-specific N2 fixation rates, calculated as 15N2 incorporation rate divided by specific phylotype abundance (gene copy number), varied between the experiments, but generally agreed with the abundance and levels of nifH expression of each group of diazotrophs. Phosphorus additions had little effect on the abundance, gene expression, or N2 fixation activity of diazotrophs across the course of the 36-h experiments, indicating that other factors may limit the growth, gene expression, and activity of diazotrophs in the North Pacific subtropical gyre. [ABSTRACT FROM AUTHOR]

The availability of nitrogen is important in regulating biological productivity in marine environments. Deepwater nitrate has long been considered the major source of new nitrogen supporting primary production in oligotrophic regions of the open ocean, but recent studies have showed that biological N2 fixation has a critical role in supporting oceanic new production. Large colonial cyanobacteria in the genus Trichodesmium and the heterocystous endosymbiont Richelia have traditionally been considered the dominant marine N2 fixers, but unicellular diazotrophic cyanobacteria and bacterioplankton have recently been found in the picoplankton and nanoplankton community of the North Pacific central gyre, and a variety of molecular and isotopic evidence suggests that these unicells could make a major contribution to the oceanic N budget. Here we report rates of N2 fixation by these small, previously overlooked diazotrophs that, although spatially variable, can equal or exceed the rate of N2 fixation reported for larger, more obvious organisms. Direct measurements of 15N2 fixation by small diazotrophs in various parts of the Pacific Ocean, including the waters off Hawaii where the unicellular diazotrophs were first characterized, show that N2 fixation by unicellular diazotrophs can support a significant fraction of total new production in oligotrophic waters.