The heterocystous cyanobacteria are currently placed in subsections IV and V, which are distinguished by cellular division in one plane (false branching) and in more than one plane (true branching), respectively. Published phylogenies of 16S rRNA gene sequence data support the monophyly of the heterocystous cyanobacteria, with members of subsection V embedded within subsection IV. It has been postulated that members of subsection V arose from within subsection IV. Therefore, phylogenetic analysis of nucleotide sequences of the nitrogen-fixation gene nifD from representatives of subsections IV and V was performed by using maximum-likelihood criteria. The heterocystous cyanobacteria are supported as being monophyletic, with the non-heterocystous cyanobacteria as their closest relative. However, neither subsection IV nor subsection V is monophyletic, with representatives of both subsections intermixed in two sister clades. Analysis of nifD does not support recognition of two distinct subsections.Cyanobacteria are oxygenic, photosynthetic prokaryotes that can be found in almost every aquatic and terrestrial environment (Castenholz & Waterbury, 1989) and are ancient organisms that date back 3?5 billion years in the fossil record (Castenholz, 1992). It has been proposed that cyanobacteria were responsible for converting the ancient Earth's anaerobic atmosphere to an aerobic one (Hayes, 1983;Schopf et al., 1983). Many cyanobacteria fix nitrogen (diazotrophy) and they comprise one of the largest global suppliers of fixed nitrogen in the environment (Sprent & Sprent, 1990). In addition, some species of nitrogen-fixing cyanobacteria are involved in symbiotic relationships with many plant species and supply the host directly with a source of reduced nitrogen.The taxonomy of the cyanobacteria has been debated vigorously and revised many times (Golubic, 1976;Turner, 1997). The cyanobacteria have been reclassified by using bacteriological instead of botanical criteria, based on morphological and developmental features, but most species, including those used in this study, have not yet been given validly published names under the Bacteriological Code. Cyanobacteria are currently divided into five subsections (Rippka et al., 1979;Rippka, 1988;Castenholz & Waterbury, 1989;Castenholz, 1992Castenholz, , 2001Rippka & Herdman, 1992). Subsection I strains are unicellular and divide by binary fission or budding (Rippka et al., 1979). Subsection II strains are also unicellular but divide by multiple fission, resulting in the formation of baeocytes (Rippka et al., 1979;Rippka, 1988;Castenholz & Waterbury, 1989;Castenholz, 1992Castenholz, , 2001Rippka & Herdman, 1992). Subsection III strains are filamentous, non-heterocystous cyanobacteria that reproduce by trichome breakage (Rippka et al., 1979;Rippka, 1988;Castenholz & Waterbury, 1989;Castenholz, 1992Castenholz, , 2001Rippka & Herdman, 1992). Subsections IV and V are composed exclusively of heterocystous cyanobacteria, which are filamentous strains that reproduce by hormogonia formatio...