Mating-system evolution in Euplotes, from the Mendelian to a molecular approach.

Euplotes evolved multiple mating systems which have for long been assumed to be determined by "open" series of alleles at a single mat locus. Families of mating type-specific pheromones and of their mat coding genes have been extensively characterized from Euplotes species lying in different positions of Euplotes phylogenetic tree, and this characterization permitted the study of Euplotes mating systems to evolve from a Mendelian to a molecular approach. While early branching species (e.g. E. raikovi) show a mating-type determination at a single mat locus in accord with Mendelian genetics, late branching species (e.g. E. crassus) revealed, in disagreement with Mendelian genetics, a mating-type determination at two distinct mat loci implying an event of mat-gene locus duplication. One locus (orthologous) appears to be multi-allelic and deputed to synthesize pheromones distinctive of different mating types, and the second one (paralogous) deputed to synthesize pheromones that are structurally identical among different mating types

Evolution of mating systems in Euplotes

Ciliates control their sexual phenomenon of conjugation (or mating) through a genetic mechanism of mating types, which may either be only two within a species (recalling the duality of sexes in animals), or multiple (recalling self/non-self compatibility systems in plants and fungi). The nearly one hundred species of the most ubiquitously distributed ciliate, Euplotes, all evolved multiple mating types. Based on analyses of Mendelian genetics, these mating types have for long been assumed to be determined by multi-allelic series of genes inherited at a single genetic locus (i.e., the mating-type or mat locus) and responsible for the synthesis of mating type-pecific signaling proteins. The chemical characterization of these signaling proteins (known as pheromones) from an array of Euplotes species has now permitted us to evolve in the study of Euplotes mating types from an approach of Mendelian genetics to an approach of molecular genetics. In this new experimental context, we have cloned and characterized structurally the pheromone (mating-type) gene families of Euplotes species that take different positions in the phylogenetic tree of the genus Euplotes. It appeared that, in accord with the prediction of the Mendelian genetics, early branching species (e.g., E. polaris, E. raikovi and E. nobilii) inherit their mating types at a single multi-allelic locus. However, in disagreement with the prediction of the Mendelian genetics, late branching species (e.g., E. crassus and E. focardii) inherit their mating types at two distinct loci that are likely the result of an event of gene duplication in the germinal (micronuclear) genome. One locus appears to be structurally and functionally homologous with the multi-allelic locus of the early branching species, while the second locus appears to be structurally homologous but functionally divergent

Antarctic and Arctic populations of the ciliate Euplotes nobilii show common pheromone-mediated cell-cell signaling and cross-mating.

Wild-type strains of the protozoan ciliate Euplotes collected from different locations on the coasts of Antarctica, Tierra del Fuego and the Arctic were taxonomically identified as the morpho-species Euplotes nobilii, based on morphometric and phylogenetic analyses. Subsequent studies of their sexual interactions revealed that mating combinations of Antarctic and Arctic strains form stable pairs of conjugant cells. These conjugant pairs were isolated and shown to complete mutual gene exchange and cross-fertilization. The biological significance of this finding was further substantiated by demonstrating that close homology exists among the threedimensional structures determined by NMR of the water-borne signaling pheromones that are constitutively secreted into the extracellular space by these interbreeding strains, in which these molecules trigger the switch between the growth stage and the sexual stage of the life cycle. The fact that Antarctic and Arctic E. nobilii populations share the same gene pool and belong to the same biological species provides new support to the biogeographic model of global distribution of eukaryotic microorganisms, which had so far been based exclusively on studies of morphological and phylogenetic taxonomy

Immunotoxicity and Sensitizing Capacity of Metal Compounds Depend on Speciation

Immunotoxicity of metal compounds is an issue of great importance due to the recent industrial application of metals with unknown toxicity on the immune system and the discovery of metal intermediary compounds not sufficiently studied yet. In this report we show results of our study on the immunotoxicity of the following metals: the Platinum group elements (Platinum, Palladium, Rhodium), Titanium and Arsenic. We applied functional and non functional assays and investigated both innate and adaptive immune systems, in particular, cell proliferation, cytokine production by PBMCs and O−2 production by neutrophils. We obtained the following results: only some Ti compounds (Titanocene, Ti ascorbate and Ti oxalate) show immunotoxicity. Trivalent As compounds (Sodium arsenite and tetraphenyl arsonium chloride) are more immunotoxic than the other investigated As compounds. Genotoxicity of Pt group compounds is in the following order: Pt < Rh < Pd. Immunotoxicity of Pt group compounds is in the following order: Pd < Pt < Rh. Lymphocytes and macrophages show a different reaction of neutrophils to metal toxicity. We can conclude that these studies show that metal immunotoxicity depends on speciation. In general speciation provides additional and often essential information in evaluating metal toxicity. However, there are many difficulties in applying speciation in investigating toxico-kinetic aspects to many metals, mainly due to the lack of information about the existence and significance of species and to the lack of analytical methods for measuring species in biological samples

Evidence for Gene Duplication and Allelic Codominance (not Hierarchical Dominance) at the Mating-Type Locus of the Ciliate, Euplotes crassus.

The high-multiple mating system of Euplotes crassus is known to be controlled by multiple alleles segregating at a single locus and manifesting relationships of hierarchical dominance, so that heterozygous cells would produce a single mating-type substance (pheromone). In strain L-2D, now known to be homozygous at the mating-type locus, we previously identified two pheromones (Ec-α and Ec-1) characterized by significant variations in their amino acid sequences and structure of their macronuclear coding genes. In this study, pheromones and macronuclear coding genes have been analyzed in strain POR-73 characterized by a heterozygous genotype and strong mating compatibility with L-2D strain. It was found that POR-73 cells contain three distinct pheromone coding genes and, accordingly, secrete three distinct pheromones. One pheromone revealed structural identity in amino acid sequence and macronuclear coding gene to the Ec-α pheromone of L-2D cells. The other two pheromones were shown to be new and were designated Ec-2 and Ec-3 to denote their structural homology with the Ec-1 pheromone of L-2D cells. We interpreted these results as evidence of a phenomenon of gene duplication at the E. crassus mating-type locus, and lack of hierarchical dominance in the expression of the macronuclear pheromone genes in cells with heterozygous genotypes

A Chance-Constrained Linear Model for Beef Policy Analysis

A simple linear model to evaluate different bull beef policies (BBP) by incorporating the stochastic nature of pasture yield from a database with separate chance constraints (ChC) was developed. A 100 ha farm was used, and the model was run with five different BBP and set to different levels of pasture yield risk to maximise production.ha-1.year-1. Summer was the most risky season, and the optimum mix of policies changed at different levels of risk. Chance constraint represents an interesting and simple alternative to include pasture variation into a linear model

Effects of Herbage Allowance upon Animal Performance and Grazing Behavior of Strip-Grazed Heifers

A trial was designed to test the effect of herbage allowance (HA) on live-weight gain (LWG) and grazing behavior of heifers during spring as part of a combined field-modeling research programme. Low HA (L) and high HA (H) of 2.5 and 5.0 kg DM herbage mass 100 kg animal LW-1day-1 were fed respectively. H animals grazed longer and achieved a higher LWG than L (P \u3c 0.05). H animals left a higher residual pasture (P \u3c 0.051) with a significantly (P \u3c 0.05) higher content of green, clover, non-lamina and petiole of a higher digestibility and NSC, with a lower NDF content. The significance of some factors involved in these results on intake and diet selection are discussed in relation to the predictability of animal performance