Morphology and phylogeny of a new woodruffiid ciliate, Etoschophrya inornata sp. n. (Ciliophora, Colpodea, Platyophryida), with an account on evolution of platyophryids

Abstract

We studied the morphology, morphometry, resting cysts, and molecular phylogeny of a new woodruffiid ciliate, Etoschophrya inornata, from ephemeral puddles and two lacustrine habitats in Idaho, Northwest U.S.A. Up to now the genus Etoschophrya has included a single species, E. oscillatoriophaga, from which our new form is distinguished by (1) the absence of interkinetal cortical granules and, consequently, the absence of extrusible red material in methyl green-pyronin stains; (2) usually ≥ five adoral membranelles vs. usually four; (3) greater length and length:width ratio; (4) prominent cortical furrows vs. inconspicuous; and (5) adaptation to nonsaline semiterrestrial and lacustrine habitats in the Nearctic vs. highly saline alkaline Afrotropic soil habitats. Resting cysts have two distinct membranes and a thick hyaline mucous pericyst layer. However, only one membrane persists in older cysts. Like its congener, E. inornata feeds exclusively on filamentous cyanobacteria. The 18S rRNA gene sequence places this species in a strongly supported clade with Kuklikophrya ougandae basal to the other platyophryids. We include a morphologic cladistic analysis of platyophryid ciliates and present a hypothetical scenario for the evolution of the platyophryid oral structures.

Phylogeny of the ciliate family Psilotrichidae (Protista, Ciliophora), a curious and poorly-known taxon, with notes on two algae-bearing psilotrichids from Guam, USA

“…PCR was done as follows: in 0.2 ml PCR tubes, we used 12.5 μL GoTaq® Green Master Mix, 2× (Madison, WI, USA), 1.25 μl each of universal eukaryotic forward primer EUK-A and reverse primer EUK-B [46], both in a final concentration of 0.4 μM, and 10.5 μl of the Chelex extraction for a total volume of 25 μl. PCR was performed in the iCycler™ Thermal Cycler and DNA sequencing in both directions was done at GENEWIZ (South Plainfield, NJ, USA) as previously described [47]. Contigs were assembled by Seqman (DNAStar).…”

Placing Environmental Next-Generation Sequencing Amplicons from Microbial Eukaryotes into a Phylogenetic Context

“…Although the Colpodea are recognized as a taxon based on the presence of the LKm (left kinetodesmal) fiber in the somatic ciliature (Lynn 1976;Small and Lynn 1981;Foissner 1993), molecular support for monophyly from nuclear SSU-rDNA data was lacking in an earlier study when all potentially closely related outgroups were included (Dunthorn et al 2008). Later nuclear and mictochondrial SSU-rDNA analyses did not sample sufficient outgroups to allow for a meaningful test of monophyly (Lynn et al 1999;Lasek-Nesselquist and Katz 2001;Dunthorn et al 2008Dunthorn et al , 2009Foissner and Stoeck 2009;Bourland et al 2011;Dunthorn et al 2011;Foissner et al 2011;Quintela-Alonso et al 2011;Bourland et al 2012;Dunthorn, Katz, et al 2012;Bourland et al 2013;Foissner et al 2013). With the increased taxon sampling here, the Colpodea are inferred to be monophyletic with PairDist, RAxML, and MrBayes; we obtain high bipartition support for this relationship only from PairDist and MrBayes.…”

“…Second, they may be visually unremarkable and easily missed in morphological studies that focused on more charismatic ciliates such as in the Karyorelictea, Oligohymenophorea, and Spirotrichea. Third, those researchers studying the colpodeans primarily focus on terrestrial environments (e.g., Foissner 1993;Dunthorn et al 2008;Quintela-Alonso et al 2011;Bourland et al 2012). Fourth, the colpodeans amplified and pyrosequenced from the marine samples may not actually be normally metabolically active in the sampling sites and were merely from recent continental runoff waters.…”

Morphologic and molecular description of Metopus fuscus Kahl from North America and new rDNA sequences from seven metopids (Armophorea, Metopidae)

“…DNA was extracted from each of four cells using a modified Chelex method (Strüder-Kypke and Lynn 2003) and the 18S rDNA was amplified and sequenced as previously described (Bourland et al 2012). Chromatograms were manually edited using 4-Peaks (Griekspoor and Groothuis 2006) and assembled into contiguous sequences using CAP3 (Huang and Madan 1999).…”