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Marine microbes are an essential component of global biogeochemical cycles. In oligotrophic marine surface waters, the phytoplankton, phototrophic, single-celled (on occasion, colonial) organisms, is often dominated by the picoplankton (cells <2 micrometers in size), which constitute the base of the marine food chain. The picophytoplankton is composed of three main groups of organisms: two genera of cyanobacteria, Prochlorococcus and Synechococcus, and a third group, the picoeukaryotes. Even though numerically less abundant than cyanobacteria, picoeukaryotes can contribute significantly to biomass and primary production in this size fraction. Furthermore, picoeukaryotes are a diverse group but this diversity is still underexplored and their ecological roles and physiology is poorly understood. Here uncultured protists are investigated using 18S rRNA gene clone libraries, phylogenetic analyses, specific fluorescence in situ hybridization (FISH) probes and other methods in tropical and subtropical waters. Gene sequences comprising a unique eukaryotic lineage, biliphytes, were identified in most samples, whether from high (30 degrees Celsius) or low (5 degrees Celsius) temperature waters. Sequences within this uncultured group have previously been retrieved from mid and high latitudes. Phycobilin-like fluorescence associated with biliphyte-specific FISH probed cells indicated they may be photosynthetic. Furthermore, the data indicated biliphytes are nanoplanktonic in size, averaging between 3.0 and 4.1 micrometers. Using the 18S rRNA gene, sequences belonging to a broadly distributed but uncultivated pico-prymnesiophytes were retrieved. We investigated the ecological importance of these natural pico-prymnesiophyte populations and field experiments showed that they could grow rapidly and contributed measurably to primary production. They also appear to form a large portion of global picophytoplankton biomass, with differing contributions in five biogeographical provinces, from tropical to high latitudes. Finally, the physiology of the picoeukaryote Micromonas was studied under a shift from medium to high light and UV radiation. Results showed that the growth of these photosynthetic cells was synchronized with the light: dark period. Forward angle side scatter and red autofluorescence from chlorophyll increased throughout the light period and decreased during the dark period. This is consistent with cell division occurring at the beginning of the dark period. Additionally, genes proposed to have roles in photoprotection were up-regulated under high light and UV, but not in controls.