Genealogical analyses of multiple loci of litostomatean ciliates (Protista, Ciliophora, Litostomatea)

© The Author(s), 2012. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in Molecular Phylogenetics and Evolution 66 (2012): 397-411, doi:10.1016/j.ympev.2012.06.024.The class Litostomatea is a highly diverse ciliate taxon comprising hundreds of free-living and endocommensal species. However, their traditional morphology-based classification conflicts with 18S rRNA gene phylogenies indicating (1) a deep bifurcation of the Litostomatea into Rhynchostomatia and Haptoria + Trichostomatia, and (2) body polarization and simplification of the oral apparatus as main evolutionary trends in the Litostomatea. To test whether 18S rRNA molecules provide a suitable proxy for litostomatean evolutionary history, we used eighteen new ITS1-5.8S rRNA-ITS2 region sequences from various free-living litostomatean orders. These single- and multiple-locus analyses are in agreement with previous 18S rRNA gene phylogenies, supporting that both 18S rRNA gene and ITS region sequences are effective tools for resolving phylogenetic relationships among the litostomateans. Despite insertions, deletions and mutational saturations in the ITS region, the present study shows that ITS1 and ITS2 molecules can be used to infer phylogenetic relationships not only at species level but also at higher taxonomic ranks when their secondary structure information is utilized to aid alignment.Financial support was provided by the Austrian Science Foundation (FWF Projects P-19699-B17 and P-20360-B17 to Wilhelm Foissner), the Slovak Scientific Grant Agency (VEGA Project 1/0600/11 to Peter Vd’ačný), and US NSF Grants (Projects MCB-0348341 and DEB-0816840 to Slava S. Epstein)

Protistan community patterns within the brine and halocline of deep hypersaline anoxic basins in the eastern Mediterranean Sea

Author Posting. © The Author(s), 2008. This is the author's version of the work. It is posted here by permission of Springer for personal use, not for redistribution. The definitive version was published in Extremophiles 13 (2009): 151-167, doi:10.1007/s00792-008-0206-2.Environmental factors restrict the distribution of microbial eukaryotes but the exact boundaries for eukaryotic life are not known. Here we examine protistan communities at the extremes of salinity and osmotic pressure, and report rich assemblages inhabiting Bannock and Discovery, two deep-sea superhaline anoxic basins in the Mediterranean. Using a rRNA-based approach, we detected 1538 protistan rRNA gene sequences from water samples with total salinity ranging from 39 g/kg to 280 g/Kg, and obtained evidence that this DNA was endogenous to the extreme habitats sampled. Statistical analyses indicate that the discovered phylotypes represent only a fraction of species actually inhabiting both the brine and the brine-seawater interface, with as much as 82% of the actual richness missed by our survey. Jaccard indices (e.g., for a comparison of community membership) suggest that the brine/interface protistan communities are unique to Bannock and Discovery basins, and share little (0.8-2.8%) in species composition with overlying waters with typical marine salinity and oxygen tension. The protistan communities from the basins’ brine and brine/seawater interface appear to be particularly enriched with dinoflagellates, ciliates and other alveolates, as well as fungi, and are conspicuously poor in stramenopiles. The uniqueness and diversity of brine and brine-interface protistan communities make them promising targets for protistan discovery.This study was supported by grant grant STO414/2-4 of the Deutsche Forschungsgemeinschaft, the EuroDEEP program of the European Science Foundation under 06-EuroDEEP-FP-004 MIDDLE project and NSF-grant MCB- 034834

The Laser Astrometric Test of Relativity Mission

This paper discusses new fundamental physics experiment to test relativistic gravity at the accuracy better than the effects of the 2nd order in the gravitational field strength. The Laser Astrometric Test Of Relativity (LATOR) mission uses laser interferometry between two micro-spacecraft whose lines of sight pass close by the Sun to accurately measure deflection of light in the solar gravity. The key element of the experimental design is a redundant geometry optical truss provided by a long-baseline (100 m) multi-channel stellar optical interferometer placed on the International Space Station. The geometric redundancy enables LATOR to measure the departure from Euclidean geometry caused by the solar gravity field to a very high accuracy. LATOR will not only improve the value of the parameterized post-Newtonian (PPN) parameter gamma to unprecedented levels of accuracy of 1 part in 1e8, it will also reach ability to measure effects of the next post-Newtonian order (1/c^4) of light deflection resulting from gravity's intrinsic non-linearity. The solar quadrupole moment parameter, J2, will be measured with high precision, as well as a variety of other relativistic. LATOR will lead to very robust advances in the tests of fundamental physics: this mission could discover a violation or extension of general relativity, or reveal the presence of an additional long range interaction in the physical law. There are no analogs to the LATOR experiment; it is unique and is a natural culmination of solar system gravity experiments.Comment: 8 pages, 2 figures, invited talk given at the Second International Conference on Particle and Fundamental Physics in Space (SpacePart'03), 10-12 December 2003, Washington, D

Incubation of Environmental Samples in a Diffusion Chamber Increases the Diversity of Recovered Isolates▿ †

The majority of microorganisms from natural environments cannot be grown in the laboratory. The diffusion-chamber-based approach is an alternative method that allows microorganisms to grow in their natural environment. An inoculum is sandwiched between semipermeable (0.03-μm-pore-size) membranes of the chamber, which is then returned to the source environment. The chamber allows for a free exchange of chemicals with the external milieu by diffusion while restricting the movement of cells. We used freshwater pond sediment to inoculate diffusion chambers and petri dishes. The diffusion chambers were incubated on top of the sediment for 4 weeks. Both chamber and petri dish cultivation resulted in the isolation of numerous representatives of Alpha-, Beta-, and Gammaproteobacteria; Actinobacteria; Firmicutes; and Bacteroidetes. However, the diffusion-chamber-based approach also led to the isolation of species from rarely cultivated groups, such as Deltaproteobacteria, Verrucomicrobia, Spirochaetes, and Acidobacteria. Material from the chambers was also transferred to new chambers in order to learn whether this will increase the recovery of isolates. Several isolates could be obtained only from material transferred through multiple diffusion chambers. This suggests that continuous cultivation in diffusion chambers adapts some microorganisms for growth under otherwise prohibitive in vitro conditions

Novel Eukaryotes from the Permanently Anoxic Cariaco Basin (Caribbean Sea)

Present knowledge of microbial diversity is decidedly incomplete (S. J. Giovannoni and M. S. Rappé, p. 47-84, in D. Kirchman, ed., Microbial Ecology of the Oceans, 2000; E. Stackebrandt and T. M. Embley, p. 57-75, in R. R. Colwell and D. J. Grimes, ed., Nonculturable Microorganisms in the Environment, 2000). Protistan phylogenies are particularly deficient and undoubtedly exclude clades of principal ecological and evolutionary importance (S. L. Baldauf, Science 300:1703-1706, 2003). The rRNA approach has been extraordinarily successful in expanding the global prokaryotic record (S. J. Giovannoni and M. S. Rappé, p. 47-84, in D. Kirchman, ed., Microbial Ecology of the Oceans, 2000; E. Stackebrandt and T. M. Embley, p. 57-75, in R. R. Colwell and D. J. Grimes, ed., Nonculturable Microorganisms in the Environment, 2000) but has rarely been used in protistan discovery. Here we report the first application of the 18S rRNA approach to a permanently anoxic environment, the Cariaco Basin off the Venezuelan coast. On the basis of rRNA sequences, we uncovered a substantial number of novel protistan lineages. These included new clades of the highest taxonomic level unrelated to any known eukaryote as well as deep branches within established protistan groups. Three novel lineages branch at the base of the eukaryotic evolutionary tree preceding, contemporary with, or immediately following the earliest eukaryotic branches. These newly discovered protists may retain traits reminiscent of an early eukaryotic ancestor(s)