The acalyptrate fly superfamily Opomyzoidea, as currently recognized, is a poorly-known group of 14 families. The composition of this group and relationships among included families have been controversial. Furthermore, the delimitation of two opomyzoid families, Aulacigastridae and Periscelididae, has been unstable with respect to placement of the genera Stenomicra, Cyamops, and Planinasus. To test the monophyly of Opomyzoidea, previously proposed relationships between families, and the position of the three problematic genera, we sequenced over 3300bp of nucleotide sequence data from the 28S ribosomal DNA and CAD (rudimentary) genes from 29 taxa representing all opomyzoid families, as well as 13 outgroup taxa. Relationships recovered differed between analyses, and only branches supporting well-established monophyletic families were recovered with high support, with a few exceptions. Opomyzoidea and its included subgroup, Asteioinea, were found to be non-monophyletic. Stenomicra, Cyamops, and Planinasus group consistently with Aulacigastridae, contrary to recent classifications. Xenasteiidae and Australimyzidae, two small, monogeneric families placed in separate superfamilies, were strongly supported as sister groups.
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