Aquaporins of the PIP2 class are required for efficient anther dehiscence in tobacco

Several processes during sexual reproduction in higher plants involve the movement of water between cells or tissues. Before flower anthesis, anther and pollen dehydration takes place before the release of mature pollen at dehiscence. Aquaporins represent a class of proteins that mediates the movement of water over cellular membranes. Aquaporins of the plasmamembrane PIP2 family are expressed in tobacco (Nicotiana tabacum) anthers and may therefore be involved in the movement of water in this organ. To gain more insight into the role these proteins may play in this process, we have analyzed their localization using immunolocalizations and generated plants displaying RNA interference of PIP2 aquaporins. Our results indicate that PIP2 protein expression is modulated during anther development. Furthermore, in tobacco PIP2 RNA interference plants, anther dehydration was slower, and dehiscence occurred later when compared with control plants. Together, our results suggest that aquaporins of the PIP2 class are required for efficient anther dehydration prior to dehiscence.     

The reaction between phosphatidylethanolamines and HOCl investigated by TLC: fading of the dye primuline is induced by dichloramines

Phosphatidylethanolamines (PEs) and phosphatidylglycerols (PGs) are abundant lipid constituents of the membranes of Escherichia coli. The reaction between these lipids and hypochlorous acid (HOCl), an important constituent of disinfectants, was investigated by combined thin-layer chromatography (TLC), mass spectrometry (MS), UV and fluorescence spectroscopy. Primuline is a common dye in lipid research that binds non-covalently to lipids and allows, thus, the direct evaluation of TLC plates by MS. However, primuline staining of the products between PE and HOCl is accompanied by fading of the dye. This only holds if acidic but not alkaline conditions are applied. Using a combination of TLC, UV and fluorescence spectroscopy, it will be shown that dichloramines of PE are responsible for the observed primuline fading. Since dichloramines are slowly converted under alkaline conditions into the nitriles that lack the characteristic UV properties of dichloramines, fading is not observed under alkaline conditions.     

Saccharomyces cerevisiae wine strains differing in copper resistance exhibit different capability to reduce copper content in wine

Two wine strains of Saccharomyces cerevisiae, characterized by a different degree of copper resistance, were tested in grape must fermentation in the presence of different copper concentrations. The sensitive strain SN9 was strongly affected by copper concentration (32 ppm, (32 mg/l)), whereas the resistant strain SN41 exhibited a good growth activity in presence of 32 ppm of copper and only a reduced activity in presence of 320 ppm. The different strain fermentation performance in response to the copper addition corresponded to a different capability to accumulate copper inside the cells. Both strains exhibited the capacity to reduce the copper content in the final product, eventhough a significantly greater reducing activity was exerted by the resistant strain SN41, which was able to reduce by 90% the copper concentration in the final product and to accumulate the metal in great concentrations in the cells. As high concentrations of copper can be responsible for wine alterations, the selection of S. cerevisiae strains possessing high copper resistance and the ability to reduce the copper content of wine has a great technological interest, in particular for the fermentation of biological products. From the results obtained, the technique proposed is not only suitable for the assay of copper residues in must, wine and yeast cells, but it also offers the advantage of easy sample preparation and low detection limit in the ppb (g/l) range.     

Pollinator Competition as a Driver of Floral Divergence: An Experimental Test

Optimal foraging models of floral divergence predict that competition between two different types of pollinators will result in partitioning, increased assortative mating, and divergence of two floral phenotypes. We tested these predictions in a tropical plant-pollinator system using sexes of purple-throated carib hummingbirds (Anthracothorax jugularis) as the pollinators, red and yellow inflorescence morphs of Heliconia caribaea as the plants, and fluorescent dyes as pollen analogs in an enclosed outdoor garden. When foraging alone, males exhibited a significant preference for the yellow morph of H. caribaea, whereas females exhibited no preference. In competition, males maintained their preference for the yellow morph and through aggression caused females to over-visit the red morph, resulting in resource partitioning. Competition significantly increased within-morph dye transfer (assortative mating) relative to non-competitive environments. Competition and partitioning of color morphs by sexes of purple-throated caribs also resulted in selection for floral divergence as measured by dye deposition on stigmas. Red and yellow morphs did not differ significantly in dye deposition in the competition trials, but differences in dye deposition and preferences for morphs when sexes of purple-throated caribs foraged alone implied fixation of one or the other color morph in the absence of competition. Competition also resulted in selection for divergence in corolla length, with the red morph experiencing directional selection for longer corollas and the yellow morph experiencing stabilizing selection on corolla length. Our results thus support predictions of foraging models of floral divergence and indicate that pollinator competition is a viable mechanism for divergence in floral traits of plants.     

Low-potential sensitive H2O2 detection based on composite micro tubular Te adsorbed on platinum electrode

In this work a new original amperometric sensor for H(2)O(2) detection based on a Pt electrode modified with Te-microtubes was developed. Te-microtubes, synthesized by the simple thermal evaporation of Te powder, have a tubular structure with a hexagonal cross-section and are open ended. Modified electrode was prepared by direct drop casting of the mixture of Te-microtubes dispersed in ethanol on Pt surface. The spectroscopic characterization of synthesized Te-microtubes and Pt/Te-microtubes modified electrodes was performed by scanning electron microscopy (SEM), energy-dispersive X-rays microanalysis (EDX), X-ray diffraction analysis (XRD) and X-ray photoelectron spectroscopy (XPS). Moreover a complete electrochemical characterization of the new composite material Pt/Te-microtubes was performed by cyclic voltammetry (CV) and cronoamperometry (CA) in phosphate buffer solution (PBS) at pH 7. Electrochemical experiments showed that the presence of Te-microtubes on modified electrode was responsible for an increment of both cathodic and anodic currents in presence of H(2)O(2) with respect to bare Pt. Specifically, data collected from amperometric experiments at -150 mV vs. SCE in batch and -200 mV vs. SCE in flow injection analysis (FIA) experiments show a remarkable increment of the cathodic current. The electrochemical performances of tested sensors make them suitable for the quantitative determination of H(2)O(2) substrate both in batch and in FIA.     

An Evolutionary Analysis of Antigen Processing and Presentation across Different Timescales Reveals Pervasive Selection

The antigenic repertoire presented by MHC molecules is generated by the antigen processing and presentation (APP) pathway. We analyzed the evolutionary history of 45 genes involved in APP at the inter- and intra-species level. Results showed that 11 genes evolved adaptively in mammals. Several positively selected sites involve positions of fundamental importance to the protein function (e.g. the TAP1 peptide-binding domains, the sugar binding interface of langerin, and the CD1D trafficking signal region). In CYBB, all selected sites cluster in two loops protruding into the endosomal lumen; analysis of missense mutations responsible for chronic granulomatous disease (CGD) showed the action of different selective forces on the very same gene region, as most CGD substitutions involve aminoacid positions that are conserved in all mammals. As for ERAP2, different computational methods indicated that positive selection has driven the recurrent appearance of protein-destabilizing variants during mammalian evolution. Application of a population-genetics phylogenetics approach showed that purifying selection represented a major force acting on some APP components (e.g. immunoproteasome subunits and chaperones) and allowed identification of positive selection events in the human lineage.We also investigated the evolutionary history of APP genes in human populations by developing a new approach that uses several different tests to identify the selection target, and that integrates low-coverage whole-genome sequencing data with Sanger sequencing. This analysis revealed that 9 APP genes underwent local adaptation in human populations. Most positive selection targets are located within noncoding regions with regulatory function in myeloid cells or act as expression quantitative trait loci. Conversely, balancing selection targeted nonsynonymous variants in TAP1 and CD207 (langerin). Finally, we suggest that selected variants in PSMB10 and CD207 contribute to human phenotypes. Thus, we used evolutionary information to generate experimentally-testable hypotheses and to provide a list of sites to prioritize in follow-up analyses.