With the aim to explore the diversity of aquatic fungi in Mexico we present an investigation using a fragment of the 18S ribosomal DNA as a molecular marker obtained from different water bodies (marine, brackish and fresh water). Ribosomal gene fragments were obtained by DNA amplification, the resulting sequences were compared using multiple alignments against a collection of classified reference fungal sequences and then subjected to phylogenetic clustering allowing the identification and classification of DNA sequences from environmental isolates as fungal down to the family level, provided enough reference sequence were available. From our ensemble of 2,020 sequences identified as fungal, 23.8% were classified at the family level, 48.5% at the order level, 13% at the class/subphylum level and 14.7% of the sequences (all from the same site) could not be unambiguously positioned in any of our reference fungal groups but were closely related to uncultivated marine fungi. The most frequently recovered phylum was Ascomycota (89.1%), followed by Chytridiomycota (8.1%), Basidiomycota (2.8%) and Mucoromycotina (1.3%).