ABSTRACT. Three complete 18s ribosomal RNA gene sequences from the rumen ciliates, Entodinium caudatum (1,639 bp), Epidinium caudatum (1,638 bp), and Polyplastron multivesiculatum (1,640 bp) were determined and confirmed in the opposite direction. Trees produced using maximum parsimony and distance-matrix methods (least squares and neighbour-joining), with strong bootstrap support, depict the rumen ciliates as a monophyletic group. Entodinium caudatum is the earliest branching rumen ciliate. However, Entodiniuin simplex does not pair with En. caudatum, but rather with Polyplastron multivesiculatum. Signature sequences for these rumen ciliates reveal that the published SSrRNA gene sequence from En. simplex is in fact a Polyplastron species. The free-living haptorian ciliates, Loxophyllum, Homalozoon and Spathidium (Subclass Haptoria), are monophyletic and are the sister group to the rumen ciliates. The litostomes (Class Litostomatea), consisting of the haptorians and the rumen ciliates, are also a monophyletic group. The rumen ciliates are characterized by generally having: (1) unspecialized oral ciliature, (2) a somatic kinetid that is typically made up of one kinetosome with usually two transverse microtubular ribbons evident only during kinetosomal replication, and ( 3 ) microtubular bundles (nematodesmata) that extend into the cytoplasm from the bases of kinetids that surround the cytostome [38]. At present, there are two major classification schemes for the Phylum Ciliophora, based primarily upon ultrastructural features of the ciliate cortex. For de Puytorac and his collaborators [8-lo], the rumen ciliates entirely comprise the Class Vestibuliferea, with three orders (Trichostomatida, Entodiniomorphida and Blepharocorythida), and are a sister group To whom correspondence should be addressed. Telephone: 519-824-4120, ext. 2975; Fax: 519-767-1656; Email: awright@uoguelph.ca to the Class Litostomatea within the Subphylum Filicorticata.Lynn and his collaborators [38, 48, 491 recognize two orders of rumen ciliates, Vestibuliferida and Entodiniomorphida, within the Subclass Trichostomatia, a sister group to the Subclass Haptoria within the Class Litostomatea. Despite the differences in rank for the rumen ciliates, both schemes are basically quite similar. Molecular data will be useful in testing these proposed relationships.Therefore, we sequenced in both directions the complete small subunit ribosomal RNA (SSrRNA) genes from Entodinium caudatum, Epidinium caudatum, and Polyplastron multivesiculatum and compared these to SSrRNA gene sequences from other ciliates and eukaryotes. Phylogenetic analyses were used to determine (1) if the rumen ciliates form a monophyletic group, (2) if En. caudatum and En. simplex form a clade basal to the other two entodiniomorphids, (3) if the rumen ciliates are the sister group to the free-living haptorian ciliates represented by Homalozoon, Loxophyllum, and Spathidium, and (4) if the Class Litostomatea, to which the rumen ciliates belong, is monophyletic.
MATERIALS AND METHODSS...