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Protistology ■ 65
jirovecii antibodies in sera. In order to obtain the highest amount of pure antigen, the expression vector pLATE 31, which contains the coding sequence for the MSG g antigen, was isolated and cloned in E. coli XJb (DE3). Extraction and purification through affinity chromatography with immobilized metallic ions, ELISA, SDS-PAGE and Western-Blot, were performed with the objective to obtain the maximum quantity of antigen and determine its purity. The antigen was then used to form bionanoconjugates with AuNPs, previously functionalized with several types of ligands, which will be the core of the immunochromatographic RDT with potential for point-of-care diagnostics. Acknowledgments: Partially supported by Gilead GENESE-PGG/001/2014.
CHANGES IN CILIATES COMMUNITY ACROSS AN EUTROPHICATION GRADIENT IN A LARGE TEMPERATE ESTUARY Rubin E., Santoferrara L., Zhang H., McManus G. Department of Marine Sciences, University of Connecticut, Groton, CT, USA ewelina.rubin@uconn.edu
The ciliate community composition in Long Island Sound, USA, was investigated using high throughput sequencing (Miseq, Illumina) ofvariable region 4 (V4) of the small subunit ribosomal gene (V4-SSU). Due to its proximity to heavily populated areas, the western Sound (WLIS) is eutrophic, and strong gradients have been observed down-estuary (east to west) in nutrients, chlorophyll, and plankton biomass. We sampled six stations on the long axis of the estuary, at two depths (surface and 6 m), in the summer of 2015. Analysis of ciliate V4-SSU sequences grouped into operational taxonomic units (OTUs) corresponded to the eutrophication gradient, with higher OTU-richness found in WLIS compared to the east. The majority of ciliates were classified into Choreotrichia and Oligotrichia, contributing together 65% to 91% of all ciliate OTUs. Litostomatea and Prostomatea contributed from 4 to over 30% of the OTUs. The relative proportion of Oligotrichia was higher near the surface and increased in the eastern, more pristine stations. The Choreotrichia showed the opposite pattern. The relative proportion of Oligotrichs and Choreotrichs across the eutrophication gradient in Long Island Sound was correlated with water transparency and it could be partially explained by variation in abundance of major phytoplankton groups in the western, central and eastern Long Island Sound.
TERRESTRIAL ALGA KLEBSORMIDIUM IN THE LIGHT OF THE HYPOTHESIS „EVERYTHING IS EVERYWHERE, BUT THE ENVIRONMENT SELECTS" Rysanek D.1, Rindi F.2, Holzinger A.3, Elster J.4, Skaloud P.1
1 - Department ofBotany, Faculty ofScience, Charles University ofPrague, Benatska 2, CZ-12801, Prague, Czech Republic
2 - Dipartimento di Scienze della Vita e dell'Ambiente, Universita Politecnica delle Marche, Via Brecce Bianche, IT-60131, Ancona, Italy
3 - University of Innsbruck, Institute of Botany, Functional Plant Biology, Sternwartestrase 15, A-6020 Innsbruck, Austria
4 - Centre for PolarEcology, Faculty ofScience, University of South Bohemia, Ceske Budejovice and Academy of Sciences of the Czech Republic, Institute of Botany, Phycology Centrum, Trebon 37901, Czech Republic rysadavid@seznam.cz
Baas Becking's hypothesis "Everything is everywhere, but the environment selects" first mentioned in 1934 still contains three current controversial issues in protists: disunity in the description of diversity, distribution patterns, and the ecological preferences. We examined a very common, globally distributed terrestrial algal genus Klebsormidium and tested (i) whether there is any biogeographical pattern of this microorganism, (ii) if there is a comparable diversity in the polar and temperate regions, and (iii) a potential role of ecological speciation processes on diversification.Our analyses revealed the presence of two different distribution patterns which are supposed to characterize both macroorganisms and protists. We demonstrated an unlimited dispersal and intensive gene flow within one of the inferred lineages (superclade B). However, the majority of Klebsormidium clades showed rather a limited distribution. In addition, we detected a significant decrease of species richness towards the poles, i.e. the macro-ecological pattern typical for macroorganisms. Species within a single protist genus may thus exhibit highly contrasting distribution patterns, based on their dispersal capabilities, which are usually shaped by both intrinsic and extrinsic factors. In addition, we determined a distinct ecophysiological differentiation among distantly and closely related lineages, thereby corroborating our hypothesis that the sympatric speciation of terrestrial algae is driven by ecological divergence. We clearly showed that pH is a critical ecological factor that influences the diversity of autotrophic protists in terrestrial habitats.
