Confirmed mixed bird–insect pollination system of Scrophularia trifoliata L., a Tyrrhenian species with corolla spots

• Both bird and mixed vertebrate–insect (MVI) pollination systems are very rare in Europe and the Mediterranean region. Because MVI can ensure reproduction over a wider range of environmental conditions than when insects are the sole pollinators, under certain circumstances such systems are highly advantageous to plants. Here, we investigated the pollination and some reproductive traits of the Tyrrhenian Scrophularia trifoliata, the only species of the genus possessing two showy dark spots inside the corolla, for which MVI pollination system had been inferred on the basis of limited censuses.
• We conducted field experiments to study MVI pollination and some reproductive traits and elucidate the role of corolla spots, analysing their ultraviolet pattern, histology and pigments versus the rest of the corolla.
• The primary pollinators were wasps and passerine birds. Corolla spots absorb UV light, present abundant anthocyanins and are histologically identical to the rest of the corolla. Control flowers had higher visitation frequency than flowers without spots. S. trifoliata is self‐compatible, with efficient intrafloral protogyny and herkogamy that prevent self‐pollination but not geitonogamy.
• We confirmed the existence of a mixed bird–insect pollination system in S. trifoliata. This system is found in three other Scrophularia species with large, showy flowers – two Mediterranean (S. grandiflora and S. sambucifolia) and one Macaronesian (S. calliantha). Unlike those species, S. trifoliata has two large spots inside the corolla. These likely operate as nectar guides and their dark colouration is related to abundant anthocyanin content.

     

Stipa tenacissima as a nurse plant of the endemic species Haplophyllum bastetanum near Granada,SE Spain

Question: What is the role of Stipa tenacissima as a nurse plant of the rare and threatened endemic species Haplophyllum bastetanum? Location: Semi‐arid steppe of the Guadix‐Baza basin (SE Spain). Methods: In the two H. bastetanum populations known, we (1) estimated the number of individuals and (2) analysed the influence of the factors ‘site’ (S1 = Hernán Valle, S2 = Alicún), ‘microhabitat’ (inside and below the canopy of the S. tenacissima tussocks vs. open areas), ‘life stage’ (vegetative, reproductive), ‘aspect’ (N, S, E, W, flat) and ‘damage’ (yes or no), in relation to the distribution of individuals and to the height, number of flowers, number of capsules, and edaphic variables. Results: The total number of individuals was estimated at 3135. S. tenacissima was found to have a positive and significant effect on the distribution of H. bastetanum, more pronounced in the sample taken at S2 than S1, where 83.8% of the plants were growing in association with S. tenacissima. Similarly, there were more H. bastetanum individuals that became reproductive inside S. tenacissima tussocks than in the open areas, and H. bastetanum individuals had a greater production of flowers and fruits, especially in the sample taken at S1. Organic carbon, nitrogen, and carbon:nitrogen ratio were related to the height and number of flowers and capsules of the plants. Conclusion: Although new patches were established, with a larger number of individuals, H. bastetanum continues to be Critically Endangered (CR). The results support the hypothesis of the facilitation effect of S. tenacissima on this rare, endemic species. Facilitation of species of interest in conservation is an aspect that has not previously been clearly reported. We propound this important process as a valuable tool for rehabilitation of populations of threatened plant species in conservation biology projects.     

Identification of a novel risk locus for multiple sclerosis at 13q31.3 by a pooled genome-wide scan of 500,000 single nucleotide polymorphisms

Multiple sclerosis is a chronic inflammatory demyelinating disease of the central nervous system with an important genetic component and strongest association driven by the HLA genes. We performed a pooling-based genome-wide association study of 500,000 SNPs in order to find new loci associated with the disease. After applying several criteria, 320 SNPs were selected from the microarrays and individually genotyped in a first and independent Spanish Caucasian replication cohort. The 8 most significant SNPs validated in this cohort were also genotyped in a second US Caucasian replication cohort for confirmation. The most significant association was obtained for SNP rs3129934, which neighbors the HLA-DRB/DQA loci and validates our pooling-based strategy. The second strongest association signal was found for SNP rs1327328, which resides in an unannotated region of chromosome 13 but is in linkage disequilibrium with nearby functional elements that may play important roles in disease susceptibility. This region of chromosome 13 has not been previously identified in MS linkage genome screens and represents a novel risk locus for the disease.     

p16(INK4a) translation suppressed by miR-24

Background

Expression of the tumor suppressor p16^INK4a increases during aging and replicative senescence.

Methodology/Principal Findings

Here, we report that the microRNA miR-24 suppresses p16 expression in human diploid fibroblasts and cervical carcinoma cells. Increased p16 expression with replicative senescence was associated with decreased levels of miR-24, a microRNA that was predicted to associate with the p16 mRNA coding and 3′-untranslated regions. Ectopic miR-24 overexpression reduced p16 protein but not p16 mRNA levels. Conversely, introduction of antisense (AS)-miR-24 blocked miR-24 expression and markedly enhanced p16 protein levels, p16 translation, and the production of EGFP-p16 reporter bearing the miR-24 target recognition sites.

Conclusions/Significance

Together, our results suggest that miR-24 represses the initiation and elongation phases of p16 translation.     

Effect of epsilon toxin-GFP on MDCK cells and renal tubules in vivo.

Epsilon toxin (epsilon-toxin), produced by Clostridium perfringens types B and D, causes fatal enterotoxemia, also known as pulpy kidney disease, in livestock. Recombinant epsilon-toxin-green fluorescence protein (epsilon-toxin-GFP) and epsilon-prototoxin-GFP were successfully expressed in Escherichia coli. MTT assays on MDCK cells confirmed that recombinant epsilon-toxin-GFP retained the cytotoxicity of the native toxin. Direct fluorescence analysis of MDCK cells revealed a homogeneous peripheral pattern that was temperature sensitive and susceptible to detergent. epsilon-Toxin-GFP and epsilon-prototoxin-GFP bound to endothelia in various organs of injected mice, especially the brain. However, fluorescence mainly accumulated in kidneys. Mice injected with epsilon-toxin-GFP showed severe kidney alterations, including hemorrhagic medullae and selective degeneration of distal tubules. Moreover, experiments on kidney cryoslices demonstrated specific binding to distal tubule cells of a range of species. We demonstrate with new recombinant fluorescence tools that epsilon-toxin binds in vivo to endothelial cells and renal tubules, where it has a strong cytotoxic effect. Our binding experiments indicate that an epsilon-toxin receptor is expressed on renal distal tubules of mammalian species, including human.