Evolutionary implications of a rRNA-based phylogeny of Harpellales

The Harpellales (Trichomycetes) are endosymbiotic microfungi, mostly unculturable and predominantly associated with larval aquatic insects worldwide. Molecular phylogenies including 'gut fungi' have included at most only four axenic isolates of the 38 genera of Harpellales. Cladistic analyses were used to infer the phylogeny of the Harpellales using partial 18S or 28S nu-rRNA sequences generated for 16 genera of Harpellales, with 64 of 72 sequences generated from unculturable samples. Both analyses placed Orphella outside an otherwise monophyletic group of Harpellales, more closely allied to the Kickxellales. The current classification recognizing two families is not corroborated and continued use of the family Legeriomycetaceae may not be supportable. The largest genera of Harpellales, Smittium and Stachylina, were polyphyletic and the 28S rRNA sequences separate Smittium culisetae from the remainder of its genus. The cladograms did not support the consistent mapping of important morphological taxonomic characters, including trichospore shape and zygospore type or appendage numbers for both. This study demonstrates the use of microscopic thalli from host guts for molecular phylogenies and suggests the need for more data from the remaining Harpellales, especially with the future inclusion of protein-coding genes.