MBE Advance Access published online on September 8, 2005
Molecular Biology and Evolution, doi:10.1093/molbev/msj011
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1 Department of Biology, University of York, Heslington, York YO10 5YW, United Kingdom; Present Address: Forestry and Agricultural Biotechnology Institute, Department of Microbiology and Plant Pathology, University of Pretoria, Pretoria 0001, South Africa
* To whom correspondence should be addressed. Recent molecular studies suggest that Opisthokonta, the eukaryotic supergroup including animals and fungi, should be expanded to include a diverse collection of primitively single-celled eukaryotes previously classified as Protozoa. These taxa include corallochytreans, nucleariids, ministeriids, choanoflagellates and ichthyosporeans. Assignment of many of these taxa to Opisthokonta remains uncorroborated as it is based solely on small subunit ribosomal RNA trees lacking resolution and significant bootstrap support for critical nodes. Therefore important details of the phylogenetic relationships of these putative opisthokonts with each other and with animals and fungi, remain unclear. We have sequenced EF-1
Accepted August 30, 2005
Research Article
The Protistan Origins of Animals and Fungi
2 Department of Biology, University of York, Heslington, York YO10 5YW, United Kingdom
Sandra L. Baldauf, E-mail: slb14{at}york.ac.uk
Abstract 
, actin, 
-tubulin, HSP70 and/or -tubulin from representatives of each of the proposed protistan opisthokont lineages, constituting the first protein-coding gene data for some of them. Our results show that members of all opisthokont protist groups encode a 
12 amino acid insertion in EF-1, previously found exclusively in animals and fungi. Phylogenetic analyses of combined multigene datasets including a diverse set of opisthokont and non-opisthokont taxa place all of the proposed opisthokont protists unequivocally in an exclusive clade with animals and fungi. Within this clade, the nucleariid appears as the closest sister taxon to fungi, while the corallochytrean and ichthyosporean form a group which, together with the ministeriid and choanoflagellates, form 2-3 separate sister lineages to animals. These results further establish Opisthokonta as a bona fide taxonomic group and suggest that any further testing of the legitimacy of this taxon should, at the least, include data from opisthokont protists. Our results also underline the critical position of these ‘animal-fungal allies’ with respect to the origin and early evolution of animals and of fungi.
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