Culture-independent surveys of marine picoeukaryotic diversity based on 18S rDNA clone libraries have unveiled numerous sequences of novel high-rank taxa. We analyzed the metagenomic dataset from the Global Ocean Survey expedition for eukaryotic ribosomal signatures. This PCR-free approach revealed similar phylogenetic patterns to clone library surveys. We also compared rDNA and rRNA 18S clone libraries from the Mediterranean Sea, revealing major differences and indicating that the rRNA approach better depicts the biogeochemical significance of marine protists.