Nitrogen fixation, apparently by bacterial endosymbionts, is associated with intertwining chains of two species of the diatom Rhizosolenia. In situ fixation rates were enhanced by incubation in the dark, whereas concurrent shipboard experiments either underestimated or did not detect nitrogen fixation. This is the first example of nitrogen fixation associated with a bacteria-diatom symbiosis in the pelagic zone, and it indicates that these systems may contribute a significant amount of "new" nitrogen to oligotrophic waters.