A single cell divides to multiply, but not all cells follow the same pattern of division. We studied cell division in seven strains from six species belonging to the genus Nannochloris Naumann and classified their modes of cell division into three types: binary fission (N. bacillaris Naumann), budding (N. coccoides Naumann), and autosporulation resulting in the formation of two to four daughter cells (N. maculata Butcher, N. sp. SAG 251-2, N. atomus Butcher CCAP 251/7 and SAG 14.87, and N. eucaryotum[Wilhelm et al.] Menzel and Wild). To determine the evolutionary relationships among these multiple modes of cell division, we investigated the strains' genome sizes, copy number of actin genes, and phylogeny. The genome sizes were determined by counter-clamped homogeneous electric fields electrophoresis and fluorimetry. The genomes are very small and range from 12.6 Mbp (N. maculata) to 47.4 Mbp (N. atomus SAG 14.87). The genomes of Nannochloris species seem to be among the smallest for free-living eukaryotes. Nannochloris bacillaris (binary fission), N. coccoides (budding), Nannochloris sp. (two-cell type of autosporulation), and N. eucaryotum (multicell type of autosporulation) contain a single actin gene, whereas N. maculata (two-cell type of autosporulation) and two strains of N. atomus (two-cell type of autosporulation) contain two actin genes. This suggests that the actin gene was duplicated in this eukaryote, which has a very small genome. Phylogenetic analyses of partial actin gene sequences suggest that autosporulation is the ancestral mode of cell division.