Web-based telemicroscopy

By taking advantage of network-based computing and the recent developments in Web interfaces, centralized research facilities housing specialized and unique imaging instruments along with associated high-performance computing can be made available to researchers for use from their own laboratories. In addition to increasing access and utilization of these facilities, operation over the Internet is expected to enhance research by facilitating collaboration between researchers. We describe the implementation of a platform-independent Web-based system written in Java that supplements automated functions with video-guided interactive, collaborative remote control and data acquisition from an intermediate-high-voltage electron microscope.     

Biosynthesis of Novel Pyoverdines by Domain Substitution in a Nonribosomal Peptide Synthetase of Pseudomonas aeruginosa

Pyoverdine is a fluorescent nonribosomal peptide siderophore made by fluorescent pseudomonads. The Pseudomonas aeruginosa nonribosomal peptide synthetase (NRPS) PvdD contains two modules that each incorporate an l-threonine residue at the C-terminal end of pyoverdine. In an attempt to generate modified pyoverdine peptides, we substituted alternative-substrate-specifying adenylation (A) and peptide bond-catalyzing condensation (C) domains into the second module of PvdD. When just the A domain was substituted, the resulting strains produced only wild-type pyoverdine—at high levels if the introduced A domain specified threonine or at trace levels otherwise. The high levels of pyoverdine synthesis observed whenever the introduced A domain specified threonine indicated that these nonnative A domains were able to communicate effectively with the PvdD C domain. Moreover, the unexpected observation that non-threonine-specifying A domains nevertheless incorporated threonine into pyoverdine suggests that the native PvdD C domain exhibited stronger selectivity than these A domains for the incorporated amino acid substrate (i.e., misactivation of a threonine residue by the introduced A domains was more frequent than misincorporation of a nonthreonine residue by the PvdD C domain). In contrast, substitution of both the C and A domains of PvdD generated high yields of rationally modified pyoverdines in two instances, these pyoverdines having either a lysine or a serine residue in place of the terminal threonine. However, C-A domain substitution more commonly yielded a truncated peptide product, likely due to stalling of synthesis on a nonfunctional recombinant NRPS template.     

Banksia born to burn

? Historical evidence of recurrent fire in many of the world's biomes suggests that fire may have had profound evolutionary influences on their extant floras. However, the role of fire as a selective force in the origin and evolution of plant traits remains controversial. ? Using Bayesian Monte-Carlo-Markov-Chain procedures and calibration points from the fossil record, we generated a dated phylogeny for the iconic Australian genus Banksia, and reconstructed the evolutionary/chronological position of five putatively fire-related traits. ? The fire-dependent trait, on-plant seed storage (serotiny), and associated fire-enhancing trait, dead floret retention, co-originated with the first appearance of Banksia 60.8 million yr ago (Palaeocene). Whether nonsprouting or resprouting is ancestral was indeterminable, but the first banksias were nonclonal. Derived traits, such as dead leaf retention (fire-enhancing) and clonality (underground budbanks; fire-avoiding), first appeared 26-16 million yr ago (Miocene) with the onset of seasonal drought and thus more frequent fire, and culminated in dead florets/bracts completely covering the persistent fruits in some species. ? Thus, fire may have been a selective force in the very origin of Banksia 40 million yr before the onset of climate seasonality in the Miocene, and continued to have an impact on the direction of evolution, favouring traits consistent with adaptation to an increasingly (sometimes less) fire-prone environment.     

Genetic control of chicken heterophil function in advanced intercross lines: associations with novel and with known <Emphasis Type="Italic">Salmonella</Emphasis> resistance loci and a likely mechanism for cell death in extracellular trap production

Heterophils, the avian polymorphonuclear leukocyte and the counterpart of mammalian neutrophils, generate the primary innate response to pathogens in chickens. Heterophil performance against pathogens is associated with host disease resistance, and heterophil gene expression and function are under genetic control. To characterize the genomic basis of heterophil function, heterophils from F₁₃ advanced intercross chicken lines (broiler × Leghorn and broiler × Fayoumi) were assayed for phagocytosis and killing of Salmonella enteritidis, oxidative burst, and extracellular trap production. A whole-genome association analysis of single nucleotide polymorphisms at 57,636 loci identified genomic locations controlling these functional phenotypes. Genomic analysis revealed a significant association of extracellular trap production with the SAL1 locus and the SLC11A1 gene, which have both been previously associated with resistance to S. enteritidis. Fine mapping supports SIVA1 as a candidate gene controlling SAL1-mediated resistance and indicates that the proposed cell-death mechanism associated with extracellular trap production, ETosis, likely functions through the CD27/Siva-1-mediated apoptotic pathway. The SLC11A1 gene was also associated with phagocytosis of S. enteritidis, suggesting that the Slc11a1 protein may play an additional role in immune response beyond depleting metal ions to inhibit intracellular bacterial growth. A region of chromosome 6 with no characterized genes was also associated with extracellular trap production. Further characterization of these novel genes in chickens and other species is needed to understand their role in polymorphonuclear leukocyte function and host resistance to disease.     

Mapping and genotypic analysis of the NK-lysin gene in chicken

Background

Antimicrobial peptides (AMP) are important elements of the first line of defence against pathogens in animals. NK-lysin is a cationic AMP that plays a critical role in innate immunity. The chicken NK-lysin gene has been cloned and its antimicrobial and anticancer activity has been described but its location in the chicken genome remains unknown. Here, we mapped the NK-lysin gene and examined the distribution of a functionally significant single nucleotide polymorphism (SNP) among different chicken inbred lines and heritage breeds.

Results

A 6000 rad radiation hybrid panel (ChickRH6) was used to map the NK-lysin gene to the distal end of chromosome 22. Two additional genes, the adipocyte enhancer-binding protein 1-like gene (AEBP1) and the DNA polymerase delta subunit 2-like (POLD2) gene, are located in the same {"type":"entrez-nucleotide","attrs":{"text":"NW_003779909","term_id":"358468969","term_text":"NW_003779909"}}NW_003779909 contig as NK-lysin, and were thus indirectly mapped to chromosome 22 as well. Previously, we reported a functionally significant SNP at position 271 of the NK-lysin coding sequence in two different chicken breeds. Here, we examined this SNP and found that the A allele appears to be more common than the G allele in these heritage breeds and inbred lines.

Conclusions

The chicken NK-lysin gene mapped to the distal end of chromosome 22. Two additional genes, AEBP1 and POLD2, were indirectly mapped to chromosome 22 also. SNP analyses revealed that the A allele, which encodes a peptide with a higher antimicrobial activity, is more common than the G allele in our tested inbred lines and heritage breeds.     

Seedling growth response to added nutrients depends on seed size in three woody genera

1 We tested whether seedlings of small‐seeded species were more reliant on soil nutrients than large‐seeded ones by growing 21 species from three woody genera ( Eucalyptus, Hakea and Banksia ) along a gradient of nutrient availability.
2 At very low nutrient availability, larger seeds produced larger seedlings. This was seen especially among the eucalypts, but the difference was eliminated at optimal soil nutrient levels. Hakea species with large seed mass, and all Banksia species, appeared unable to exploit additional soil nutrients for growth, whatever the level supplied.
3 Larger seeds tended to have proportionately higher contents of N, P and K and, under nutrient‐poor conditions, supplied more of these to their seedlings, although at a diminishing rate.
4 We suggest that large‐seededness could be an adaptation to the high‐light, nutrient‐impoverished habitats in which these species occur by providing the seedling with the mineral nutrients, rather than carbon‐based metabolites, needed for maximizing initial root growth. Reaching reliable moisture before summer (drought avoidance) is an alternative strategy to physiological tolerance of drought.     

Comparative analysis of the excretory-secretory proteome of the muscle larva of Trichinella pseudospiralis and Trichinella spiralis

The nematodes Trichinella spiralis and Trichinella pseudospiralis are both intracellular parasites of skeletal muscle cells and induce profound alterations in the host cell resulting in a re-alignment of muscle-specific gene expression. While T. spiralis induces the production of a collagen capsule surrounding the host-parasite complex, T. pseudospiralis exists in a non-encapsulated form and is also characterised by suppression of the host inflammatory response in the muscle. These observed differences between the two species are thought to be due to variation in the proteins excreted or secreted (ES proteins) by the muscle larva. In this study, we use a global proteomics approach to compare the ES protein profiles from both species and to identify individual T. pseudospiralis proteins that complement earlier studies with T. spiralis. Following two-dimensional gel electrophoresis, tandem mass spectrometry was used to identify the peptide spots. In many cases identification was aided by the determination of partial peptide sequence from selected mass ions. The T. pseudospiralis spots identified included the major secreted glycoproteins and the secreted 5'-nucleotidase. Furthermore, two major groups of T. spiralis-specific proteins and several T. pseudospiralis-specific proteins were identified. Our results demonstrate the value of proteomics as a tool for the identification of ES proteins that are differentially expressed between Trichinella species and as an aid to identifying key parasite proteins that are involved in the host-parasite interaction. The value of this approach will be further enhanced by data arising out the current T. spiralis genome sequencing project.     

Nearest-neighbour interactions in species-rich shrublands: the roles of abundance, spatial patterns and resources

We explored pairwise nearest‐neighbour interactions in four species‐rich shrubland plant communities, asking the question: how often is an individual of species j the nearest‐neighbour of species i? In the observed data and null models, less than 35% of the maximum possible number of nearest‐neighbour species pairs was present, and at three of the four sites the number of observed nearest‐neighbour pairs were significantly less than those occurring in simulated null communities. Many of the missing pairs included woody shrubs whose absence might be interpreted as evidence of site‐specific competition between larger growth forms for soil resources or space. Less than 5% of the pairs of species that occurred did so at frequencies different from that expected under random mixing, and many of these pairs were conspecific. Of the heterospecific pairs whose frequency differed significantly from random mixing there was a weak bias towards pairs occurring at higher rather than lower frequencies than expected. There was no evidence for asymmetry (interaction of species j with species i but not the reverse) in the frequency of species pairs. Nearest‐neighbour relationships are species‐specific rather than between plant functional types. The four sites form a soil nutrient and water availability gradient, and, according to the stress gradient hypothesis, positive species interactions should be most prevalent at the most stressful sites. However, we found the opposite: the site with the highest availability of resources had both proportionally the most heterospecific pairs, and the most conspecific and heterospecific species pairs with frequencies departing significantly from that expected under random mixing.     

Plant diversity in mediterranean-climate regions

The high plant diversity of mediterranean-climate regions has attracted much attention over the past few years. This review discusses patterns and determinants of local, differential and regional plant diversity in all five regions. Local diversity shows great variation within and between regions and explanations for these patterns invoke a wide range of hypotheses. Patterns of regional diversity are the result of differential speciation and extinction rates during the Quaternary. These rates have been influenced more by the incidence of fire and the severity of climate change than by environmental heterogeneity. All regions have a high number of rare and locally endemic taxa that survive as small populations, many of which are threatened by habitat transformation.