Effective control of <Emphasis Type="Italic">Botrytis</Emphasis> bunch rot in commercial vineyards by large-scale application of <Emphasis Type="Italic">Candida sake</Emphasis> CPA-1

Biological control with microorganisms is regarded as a promising control strategy against Botrytis bunch rot (BBR, grey mould) on grapes. Candida sake CPA-1 is a yeast that has previously shown a high efficacy against BBR in small-scale field trials. The present work aims to confirm this efficacy in commercial conditions and to evaluate the compatibility of C. sake with phytosanitary products that are commonly used in viticulture. The large-scale spray programmes (two to three applications in the growing season) were carried out in three field sites in Catalonia, North-East Spain, with different vineyard management styles. The overall reductions were in the ranges of 35 ± 5.7 to 64 ± 3.7% incidence, and 39 ± 17.5 to 85 ± 3.9% severity, compared to the control. Only four out of 39 phytosanitary products, tested in liquid suspension with C. sake, were incompatible with the yeast. The results demonstrated the potential of C. sake to become a reliable solution as a biocontrol product to be integrated in the control of BBR within the future.     

In cultured oligodendrocytes the A/B-type hnRNP CBF-A accompanies MBP mRNA bound to mRNA trafficking sequences

Heterogeneous ribonucleoproteins (hnRNPs) have key roles in RNA biogenesis, including pre-mRNP assembly, transport and cytoplasmic localization. Here we show by biochemical fractionation of nuclear extracts and protein-protein interaction assays that the A/B-type hnRNP CBF-A is in a multiprotein complex with hnRNP A2 and A3 and hnRNP U. Using RNA affinity chromatography and gel retardation assays, CBF-A was found to bind directly to RNA trafficking sequences in the 3'-UTR of the myelin basic protein (MBP) mRNA. In primary oligodendrocytes, astrocytes, neurons, and mouse forebrain sections, CBF-A revealed a characteristic granular cytoplasmic distribution. In mouse forebrain CBF-A-positive granules were preferentially found in regions with loosely bundled myelin fibers. In cultured oligodendrocytes, CBF-A was found to be specifically associated with endogenous MBP mRNA and CBF-A gene silencing resulted in the retention of MBP granules in the cell body. Finally, immunoelectron microscopy in differentiating oligodendrocytes showed that CBF-A is located in cytoplasmic granules that are often associated with the cytoskeleton. The results suggest that CBF-A is a novel transacting factor required for cytoplasmic mRNA transport and localization.     

Analysing the contribution of nucleic acids to the structure and properties of centric heterochromatin

A class of repetitive DNA sequences frequently found at centromeric regions are R/Y-satellites showing an asymmetric distribution of residues resulting in one strand being rich in purines (R-strand) while the complementary strand is pyrimidine-rich (Y-strand). The dodeca-satellite of Drosophila belongs to this class of centromeric satellites. In vitro, the dodeca-satellite forms altered DNA structures in which the R-strand forms very stable intramolecular fold-backs that are stabilised by the formation of tandem G · A mismatches. A single-stranded nucleic acids binding protein, DDP1, binds the unstructured dodeca-satellite Y-strand with high affinity. In polytene chromosomes, DDP1 associates with the heterochromatic chromocenter and, at the euchromatic chromosome arms, co-localises with HP1. DDP1 is a vigilin. Vigilins are highly conserved multi-KH-domain proteins. Scp160p, the vigilin from S. cerevisiae, is involved in the control of ploidy. DDP1 complements a scp160 deletion.     

Reactivity of pure Candida rugosa lipase isoenzymes (Lip1, Lip2, and Lip3) in aqueous and organic media. influence of the isoenzymatic profile on the lipase performance in organic media

Three pure isoenzymes from Candida rugosa lipase (CRL: Lip1, Lip2, and Lip3) were compared in terms of their stability and reactivity in both aqueous and organic media. The combined effect of temperature and pH on their stability was studied applying a factorial design. The analysis of the response surfaces indicated that Lip1 and Lip3 have a similar stability, lower than that of Lip2. In aqueous media, Lip3 was the most active enzyme on the hydrolysis of p-nitrophenyl esters, whereas Lip1 showed the highest activity on the hydrolysis of most assayed triacylglycerides. The highest differences among isoenzymes were found in the hydrolysis of triacylglycerides. Thus, a short, medium, and long acyl chain triacylglyceride was the preferred substrate for Lip3, Lip1, and Lip2, respectively. In organic medium, Lip3 and Lip1 provided excellent results in terms of enantioselectivity in the resolution of ibuprofen (EF value over 0.90) and conversion, whereas initial esterification rate was higher for Lip3. However, the use of Lip2 resulted in lower values of conversion, enantiomeric excess, and enantioselectivity. In the case of trans-2-phenyl-1-cyclohexanol (TPCH) resolution, initial esterification rates were high except for Lip3, which also produced poor results in conversion and enantiomeric excess. The performance of the pure isoenzymes in the enantioselectivity esterification of these substrates was compared with different CRL crude preparations with known isoenzymatic content and the different results could not be explained by their isoenzymatic profile. Therefore, it can be concluded that other factors can also affect the catalysis of CRL and only the reproducibility between powders can ensure the reproducibility in synthesis reactions.     

Colonization and diversification: towards a phylogeographic synthesis for the Canary Islands

Recently, the Canary Islands have become a focus for studies of the colonization and the diversification of different organisms. Some authors have considered Canarian endemisms as relicts of Tertiary origin, but new molecular data suggest a general pattern of continental dispersion followed by in situ speciation. Recent phylogeographic studies are revealing variants of the simple stepping-stone colonization model that seems to hold for many Hawaiian groups. Many factors can generate deviations from such a pattern: the stochastic nature of colonization, competitive exclusion, phylogenetic constraints on adaptive evolution and extinction. An understanding of island colonization and diversification can best be developed from an ecosystem level synthesis as more data for the Canarian archipelago come to hand.     

Habitat use and phenology of the large insular endemic grasshopper Acrostira euphorbiae (Orthoptera: Pamphagidae)

The habitat use and the phenology of the large grasshopper Acrostira euphorbiae García and Oromí endemic to La Palma (Canary Islands) are studied. This grasshopper is entirely dependent on the Canarian endemic shrub Euphorbia lamarckii both for food and to avoid predation. Adults stay on subapical branches during the day, probably to reduce the risk of predation, and climb up to the apex at night to feed. While females seem to ensure the genetic diversity of offspring by waiting for visits by different males, the latter have to move to guarantee their reproductive success. Monophagy in this species may be related to the year-round presence of tender shoots, and to the predator-repellent toxic latex found in the host plant. Unlike related species from continental areas, adults and nymphs of A. euphorbiae are present almost all year round, probably in adaptation to the particular climate of the islands. Nymphs are more abundant in winter, when Euphorbia leaves are most available. However, adults are more abundant than nymphs in spring, summer and autumn. Males develop more quickly than females, an apparent reproductive strategy based on achieving sexual maturity to coincide with females undergoing imaginal moulting. Matings start immediately after adult females emerge. Densities oscillated between 73 and 193 individuals ha(-1), which can be considered a low value compared with other continental pamphagid species.     

Oseltamivir-Resistant Influenza A (H1N1) Virus Strain with an H274Y Mutation in Neuraminidase Persists without Drug Pressure in Infected Mallards

Influenza A virus (IAV) has its natural reservoir in wild waterfowl, and emerging human IAVs often contain gene segments from avian viruses. The active drug metabolite of oseltamivir (oseltamivir carboxylate [OC]), stockpiled as Tamiflu for influenza pandemic preparedness, is not removed by conventional sewage treatment and has been detected in river water. There, it may exert evolutionary pressure on avian IAV in waterfowl, resulting in the development of resistant viral variants. A resistant avian IAV can circulate among wild birds only if resistance does not restrict viral fitness and if the resistant virus can persist without continuous drug pressure. In this in vivo mallard (Anas platyrhynchos) study, we tested whether an OC-resistant avian IAV (H1N1) strain with an H274Y mutation in the neuraminidase (NA-H274Y) could retain resistance while drug pressure was gradually removed. Successively infected mallards were exposed to decreasing levels of OC, and fecal samples were analyzed for the neuraminidase sequence and phenotypic resistance. No reversion to wild-type virus was observed during the experiment, which included 17 days of viral transmission among 10 ducks exposed to OC concentrations below resistance induction levels. We conclude that resistance in avian IAV that is induced by exposure of the natural host to OC can persist in the absence of the drug. Thus, there is a risk that human-pathogenic IAVs that evolve from IAVs circulating among wild birds may contain resistance mutations. An oseltamivir-resistant pandemic IAV would pose a substantial public health threat. Therefore, our observations underscore the need for prudent oseltamivir use, upgraded sewage treatment, and surveillance for resistant IAVs in wild birds.     

Overwintering of Monilinia spp. on Mummified Stone Fruit

Mummies were evaluated over the course of four growing seasons to ensure they are source of primary inoculum. The percentage of mummies with presence of conidia and its viability were determined in tree and ground mummies. The number of conidia and its germination were also quantified. Fruit mummies with Monilinia spp. were consistently detected on tree mummies in all studied orchards and growing seasons. However, the percentage of viable mummies over the same sampling periods decreased, and in most cases, it was 0% by October. The percentage of ground mummies with Monilinia spp. was lower and less viable in comparison with tree mummies, tending to decrease faster. The number of overwintering conidia in tree mummies decreased smoothly from 1 × 10⁶ to 1 × 10⁴ conidia/mummy between April and September. On the other hand, the number of conidia in ground mummies rapidly decreased to 0 conidia/mummy at around May–July. The profiles for the percentage of conidia germinated were similar in all cases. The information obtained from this study is a step forward to understanding the epidemiology of Monilinia spp., a useful tool to manage disease development.