A 2nd Generation Linkage Map of Heterobasidion annosum s.l. Based on In Silico Anchoring of AFLP Markers

In this study, we present a 2^nd generation genetic linkage map of a cross between the North American species Heterobasidion irregulare and H. occidentale, based on the alignment of the previously published 1^st generation map to the parental genomes. We anchored 216 of the original 308 AFLP markers to their respective restriction sites using an in silico-approach. The map resolution was improved by adding 146 sequence-tagged microsatellite markers and 39 sequenced gene markers. The new markers confirmed the positions of the anchored AFLP markers, fused the original 39 linkage groups together into 17, and fully expanded 12 of these to single groups covering entire chromosomes. Map coverage of the genome increased from 55.3% to 92.8%, with 96.3% of 430 markers collinearly aligned with the genome sequence. The anchored map also improved the H. irregulare assembly considerably. It identified several errors in scaffold arrangements and assisted in reducing the total number of major scaffolds from 18 to 15. This denser, more comprehensive map allowed sequence-based mapping of three intersterility loci and one mating type locus. This demonstrates the possibility to utilize an in silico procedure to convert anonymous markers into sequence-tagged ones, as well as the power of a sequence-anchored linkage map and its usefulness in the assembly of a whole genome sequence.     

Safe From Harm: Learned,Instructed, and Symbolic Generalization Pathways of Human Threat-Avoidance

Avoidance of threatening or unpleasant events is usually an adaptive behavioural strategy. Sometimes, however, avoidance can become chronic and lead to impaired daily functioning. Excessive threat-avoidance is a central diagnostic feature of anxiety disorders, yet little is known about whether avoidance acquired in the absence of a direct history of conditioning with a fearful event differs from directly learned avoidance. In the present study, we tested whether avoidance acquired indirectly via verbal instructions and symbolic generalization result in similar levels of avoidance behaviour and threat-beliefs to avoidance acquired after direct learning. Following fear conditioning in which one conditioned stimulus was paired with shock (CS+) and another was not (CS−), participants either learned or were instructed to make a response that cancelled impending shock. Three groups were then tested with a learned CS+ and CS− (learned group), instructed CS+ (instructed group), and generalized CS+ (derived group) presentations. Results showed similar levels of avoidance behaviour and threat-belief ratings about the likelihood of shock across each of the three pathways despite the different mechanisms by which they were acquired. Findings have implications for understanding the aetiology of clinical avoidance in anxiety.     

Optimising Seagrass Conservation for Ecological Functions

Ecosystems - Animals are central to numerous ecological processes that shape the structure and function of ecosystems. It follows that species that are strongly linked to specific functions can...     

Proteomics-based target identification: bengamides as a new class of methionine aminopeptidase inhibitors

LAF389 is a synthetic analogue of bengamides, a class of marine natural products that produce inhibitory effects on tumor growth in vitro and in vivo. A proteomics-based approach has been used to identify signaling pathways affected by bengamides. LAF389 treatment of cells resulted in altered mobility of a subset of proteins on two-dimensional gel electrophoresis. Detailed analysis of one of the proteins, 14-3-3gamma, showed that bengamide treatment resulted in retention of the amino-terminal methionine, suggesting that bengamides directly or indirectly inhibited methionine aminopeptidases (MetAps). Both known MetAps are inhibited by LAF389. Short interfering RNA suppression of MetAp2 also altered amino-terminal processing of 14-3-3gamma. A high resolution structure of human MetAp2 co-crystallized with a bengamide shows that the compound binds in a manner that mimics peptide substrates. Additionally, the structure reveals that three key hydroxyl groups on the inhibitor coordinate the di-cobalt center in the enzyme active site.     

Interplay of cis- and trans-regulatory mechanisms in the spliceosomal RNA helicase Brr2

RNA helicase Brr2 is implicated in multiple phases of pre-mRNA splicing and thus requires tight regulation. Brr2 can be auto-inhibited via a large N-terminal region folding back onto its helicase core and auto-activated by a catalytically inactive C-terminal helicase cassette. Furthermore, it can be regulated in trans by the Jab1 domain of the Prp8 protein, which can inhibit Brr2 by intermittently inserting a C-terminal tail in the enzyme's RNA-binding tunnel or activate the helicase after removal of this tail. Presently it is unclear, whether these regulatory mechanisms functionally interact and to which extent they are evolutionarily conserved. Here, we report crystal structures of Saccharomyces cerevisiae and Chaetomium thermophilum Brr2-Jab1 complexes, demonstrating that Jab1-based inhibition of Brr2 presumably takes effect in all eukaryotes but is implemented via organism-specific molecular contacts. Moreover, the structures show that Brr2 auto-inhibition can act in concert with Jab1-mediated inhibition, and suggest that the N-terminal region influences how the Jab1 C-terminal tail interacts at the RNA-binding tunnel. Systematic RNA binding and unwinding studies revealed that the N-terminal region and the Jab1 C-terminal tail specifically interfere with accommodation of double-stranded and single-stranded regions of an RNA substrate, respectively, mutually reinforcing each other. Additionally, such analyses show that regulation based on the N-terminal region requires the presence of the inactive C-terminal helicase cassette. Together, our results outline an intricate system of regulatory mechanisms, which control Brr2 activities during snRNP assembly and splicing.     

Dynamics ofPanonychus ulmi andTyphlodromus pyri: factors contributing to persistence

We addressed the question of persistence of predator and prey in a biological control system by examining temporal patterns ofPanonychus ulmi (Koch) and its predator,Typhlodromus pyri Scheuten at two geographic locations and at two spatial scales. At the scale of an orchard, bothP. ulmi andT. pyri were persistent over the time frame of 6 years. At the scale of an individual tree,T. pyri appeared to be more persistent than its prey,P. ulmi. We used a simulation model of single populations ofP. ulmi andT. pyri to determine which of several aspects of the biology of the two species could contribute to such a pattern. Spatial incongruity between predator and prey was essential for persistence of both species. The generalist food habit ofT. pyri probably contributes to the persistence ofT. pyri on individual trees, and may cause occasional extinction ofP. ulmi at this spatial scale. The presence of alternate food is likely an essential element for successful biological control in this system. Cannibalism byT. pyri results in higher prey densities, that is, it is detrimental to the biological control ofP. ulmi, but has no effect on the relative persistence of the two species.     

Hyphal heterogeneity in Aspergillus oryzae is the result of dynamic closure of septa by Woronin bodies

Hyphae of higher fungi are compartmentalized by septa. These septa contain a central pore that allows for inter‐compartmental and inter‐hyphal cytoplasmic streaming. The cytoplasm within the mycelium is therefore considered to be a continuous system. In this study, however, we demonstrate by laser dissection that 40% of the apical septa of exploring hyphae of Aspergillus oryzae are closed. Closure of septa correlated with the presence of a peroxisome‐derived organelle, known as Woronin body, near the septal pore. The location of Woronin bodies in the hyphae was dynamic and, as a result, plugging of the septal pore was reversible. Septal plugging was abolished in a ΔAohex1 strain that cannot form Woronin bodies. Notably, hyphal heterogeneity was also affected in the ΔAohex1 strain. Wild‐type strains of A. oryzae showed heterogeneous distribution of GFP between neighbouring hyphae at the outer part of the colony when the reporter was expressed from the promoter of the glucoamylase gene glaA or the α‐glucuronidase gene aguA. In contrast, GFP fluorescence showed a normal distribution in the case of the ΔAohex1 strain. Taken together, it is concluded that Woronin bodies maintain hyphal heterogeneity in a fungal mycelium by impeding cytoplasmic continuity.