A role for the weak DnaA binding sites in bacterial replication origins

DnaA initiates the chromosomal DNA replication in nearly all bacteria, and replication origins are characterized by binding sites for the DnaA protein (DnaA-boxes) along with an 'AT-rich' region. However, great variation in number, spatial organization and specificity of DnaA-boxes is observed between species. In the study by Taylor et al. (2011), new and unexpectedly weak DnaA-boxes were identified within the Caulobacter crescentus origin of replication (Cori). The position of weak and stronger DnaA-boxes follows a pattern seen in Escherichia coli oriC. This raises the possibility that bacterial origins might be more alike than previously thought.     

Computing diversity from dated phylogenies and taxonomic hierarchies: does it make a difference to the conclusions?

Recently, dated phylogenies have been increasingly used for ecological studies on community structure and conservation planning. There is, however, a major impediment to a systematic application of phylogenetic methods in ecology: reliable phylogenies with time-calibrated branch lengths are lacking for a large number of taxonomic groups and this condition is likely to continue for a long time. A solution for this problem consists in using undated phylogenies or taxonomic hierarchies as proxies for dated phylogenies. Nonetheless, little is known on the potential loss of information of these approaches compared to studies using dated phylogenies with time-calibrated branch lengths. The aim of this study is to ask how the use of undated phylogenies and taxonomic hierarchies biases a very simple measure of diversity, the mean pairwise phylogenetic distance between community species, compared to the diversity of dated phylogenies derived from the freely available software Phylomatic. This is illustrated with three sets of data on plant species sampled at different scales. Our results show that: (1) surprisingly, the diversity computed from dated phylogenies derived from Phylomatic is more strongly related to the diversity computed from taxonomic hierarchies than to the diversity computed from undated phylogenies, while (2) less surprisingly, the strength of this relationship increases if we consider only angiosperm species.     

Identifying indicator plant species of habitat quality and invasibility as a guide for peri-urban forest management

A floristic survey has been carried out in a peri-urban forest, the Sonian Forest in Brussels, to identify indicator plant species in the herbaceous layer, which could be used as an aid within the framework of a more sustainable management of the forest. Three hundred and seventy two (372) taxa have been identified, 33 of which are non-native (i.e. non-indigenous species regarding the study area, whether invasive or not). Criteria of habitat quality that have been chosen are the species richness, the commonness of the habitat, based on constitutive species, and its invasibility (vulnerability for invasion). On the basis of a comparison of the value of these criteria when each considered (potential indicator) species is present or not, 17 species have been recognised as reliable indicators of at least one of these three criteria. In particular, vegetation types containing either Anthriscus sylvestris, Galeopsis tetrahit or Senecio ovatus were found to be more susceptible to invasion than other habitats. The way to how the predictability of invasions might be effectively used as a management tool is discussed. Furthermore, we found a positive significant correlation (Bonferroni corrected probabilities) between the species richness and luminosity factor (derived from Ellenberg's indices), and the proportion of grassland and wetland species. The species richness was significantly negatively correlated with the proportion of woodland species. An increase in commonness was significantly correlated with a decrease in the proportion of geophytes. The usefulness of these results as an ecological basis for forest management is discussed.     

Emotional Modulation of Attention: Fear Increases but Disgust Reduces the Attentional Blink

Background

It is well known that facial expressions represent important social cues. In humans expressing facial emotion, fear may be configured to maximize sensory exposure (e.g., increases visual input) whereas disgust can reduce sensory exposure (e.g., decreases visual input). To investigate whether such effects also extend to the attentional system, we used the “attentional blink” (AB) paradigm. Many studies have documented that the second target (T2) of a pair is typically missed when presented within a time window of about 200–500 ms from the first to-be-detected target (T1; i.e., the AB effect). It has recently been proposed that the AB effect depends on the efficiency of a gating system which facilitates the entrance of relevant input into working memory, while inhibiting irrelevant input. Following the inhibitory response on post T1 distractors, prolonged inhibition of the subsequent T2 is observed. In the present study, we hypothesized that processing facial expressions of emotion would influence this attentional gating. Fearful faces would increase but disgust faces would decrease inhibition of the second target.

Methodology/Principal Findings

We showed that processing fearful versus disgust faces has different effects on these attentional processes. We found that processing fear faces impaired the detection of T2 to a greater extent than did the processing disgust faces. This finding implies emotion-specific modulation of attention.

Conclusions/Significance

Based on the recent literature on attention, our finding suggests that processing fear-related stimuli exerts greater inhibitory responses on distractors relative to processing disgust-related stimuli. This finding is of particular interest for researchers examining the influence of emotional processing on attention and memory in both clinical and normal populations. For example, future research could extend upon the current study to examine whether inhibitory processes invoked by fear-related stimuli may be the mechanism underlying the enhanced learning of fear-related stimuli.     

Effectiveness of dry heat as a seed sterilisation technique: Implications for ex situ conservation

Seed-borne pathogens are a daily issue for ex situ collection managers who try to solve it using various chemicals more or less harmful to the staff and the plant material stored. The most common physical method for seed sterilisation is moist heat, in contrast to dry heat which is used much less frequently. Consequently, the efficiency of dry heat as disinfection method and the behaviour of seeds undergoing this treatment are currently poorly known, especially for wild species. In this study, seed viability was estimated by performing germination tests according to standard procedures. Germination tests were conducted on 13,200 seeds from 66 wild species of temperate regions belonging to 22 different families. Results indicate that dry seeds exposed to 60°C during 1hour were less infected by seed-borne pathogens in 14% of the cases, whereas no change has been registered in the other cases. For all 66 studied species, no decrease in germination percentage was detected after the heat treatment. Given its positive effect on infection control without affecting seed viability, dry heat treatment as proposed here opens opportunities for seed bank managers, but also for the disinfection of herbarium collections.     

Widefield High Frame Rate Single-Photon SPAD Imagers for SPIM-FCS

Photon-counting sensors based on standard complementary metal-oxide-semiconductor single-photon avalanche diodes (SPADs) represent an emerging class of imagers that enable the counting and/or timing of single photons at zero readout noise (better than high-speed electron-multiplying charge-coupling devices) and over large arrays. They have seen substantial progress over the last 15 years, increasing their spatial resolution, timing accuracy, and sensitivity while reducing spurious signals such as afterpulsing and dark counts. They are increasingly being applied for time-resolved applications with the added advantage of enabling real-time options such as autocorrelation. We report in this study on the use of such a state-of-the-art 512 × 128 SPAD array, capable of a time resolution of 10^?5–10^?6 s for full frames while retaining acceptable photosensitivity thanks to the use of dedicated microlenses, in a selective plane illumination-fluorescence correlation spectroscopy setup. The latter allows us to perform thousands of fluorescence-correlation spectroscopy measurements simultaneously in a two-dimensional slice of the sample. This high-speed SPAD imager enables the measurement of molecular motion of small fluorescent particles such as single chemical dye molecules. Inhomogeneities in the molecular detection efficiency were compensated for by means of a global fit of the auto- and cross-correlation curves, which also made a calibration-free measurement of various samples possible. The afterpulsing effect could also be mitigated, making the measurement of the diffusion of Alexa-488 possible, and the overall result quality was further improved by spatial binning. The particle concentrations in the focus tend to be overestimated by a factor of 1.7 compared to a confocal setup; a calibration is thus required if absolute concentrations need to be measured. The first high-speed selective plane illumination-fluorescence correlation spectroscopy in vivo measurements to our knowledge were also recorded: although two-component fit models could not be employed because of noise, the diffusion of eGFP oligomers in HeLa cells could be measured. Sensitivity and noise will be further improved in the next generation of SPAD-based widefield sensors, which are currently under testing.     

Bacterial cell curvature through mechanical control of cell growth

The cytoskeleton is a key regulator of cell morphogenesis. Crescentin, a bacterial intermediate filament‐like protein, is required for the curved shape of Caulobacter crescentus and localizes to the inner cell curvature. Here, we show that crescentin forms a single filamentous structure that collapses into a helix when detached from the cell membrane, suggesting that it is normally maintained in a stretched configuration. Crescentin causes an elongation rate gradient around the circumference of the sidewall, creating a longitudinal cell length differential and hence curvature. Such curvature can be produced by physical force alone when cells are grown in circular microchambers. Production of crescentin in Escherichia coli is sufficient to generate cell curvature. Our data argue for a model in which physical strain borne by the crescentin structure anisotropically alters the kinetics of cell wall insertion to produce curved growth. Our study suggests that bacteria may use the cytoskeleton for mechanical control of growth to alter morphology.     

Germination capacity and viability of threatened species collections in seed banks

Facing the current biodiversity crisis, the value of ex situ conservation has been increasingly acknowledged in international treaties and legislations. Seed banks are a good way of conserving biodiversity, providing that seeds are of high quality and at maximum viability. However, despite the number of established ex situ facilities, there is little information on seed viability in botanic garden seed banks. This paper analyses the status of the seed collection of the National Botanic Garden of Belgium by determining the germination capacity and viability of seeds that have been stored for 1–26 years. It aims at: (1) ensuring that existing storage conditions provide effective ex situ conservation of threatened species; (2) providing viability data on threatened species; (3) planning future collection and storage efforts for seeds of West European species of conservation value. Results from this study showed that the germination and viability percentages of the 250 tested species reached on average 59 and 79% respectively. Some families typically performed better than others. Within a species, consistent results were not always obtained. Over a quarter of accessions exhibited some degree of dormancy. Considering the current lack of knowledge in seed germination and dormancy of many rare and threatened species, we believe that the quality of a seed collection should be estimated by its viability and not by its germination percentage. This study calls for further research in order to better understand the biology of a range of threatened native species.     

Patterns of native and exotic species richness in the urban flora of Brussels: rejecting the ‘rich get richer’ model

In this study we analyzed patterns of native and exotic species richness in the urban flora of Brussels (Belgium) using a coarse-scale systematic sampling grid of 1 km². The observed correlation between native and exotic richness within the grid cells sampled was then compared to the results of an adequate null model assuming no species interactions. In addition, ordinary least-squares and quantile regressions were used to analyze the relationship between the ratio of exotics to natives and the proportion of densely built up areas in each cell. Though the results obtained conform to the Eltonian expectation that exotic species preferably invade areas of low native species diversity, traditional niche-filling mechanisms seems inadequate to explain the observed pattern. Rather, aliens simply tend to have different environmental requirements than natives.     

Suppressors of DnaA(ATP) imposed overinitiation in Escherichia coli

Chromosome replication in Escherichia coli is limited by the supply of DnaA associated with ATP. Cells deficient in RIDA (Regulatory Inactivation of DnaA) due to a deletion of the hda gene accumulate suppressor mutations (hsm) to counteract the overinitiation caused by an elevated DnaA(ATP) level. Eight spontaneous hda suppressor mutations were identified by whole-genome sequencing, and three of these were analysed further. Two mutations (hsm-2 and hsm-4) mapped in the dnaA gene and led to a reduced ability to initiate replication from oriC. One mutation (hsm-1) mapped to the seqA promoter and increased the SeqA protein level in the cell. hsm-1 cells had prolonged origin sequestration, reduced DnaA protein level and reduced DnaA-Reactivating Sequence (DARS)-mediated rejuvenation of DnaA(ADP) to DnaA(ATP) , all of which could contribute to the suppression of RIDA deficiency. Despite of these defects hsm-1 cells were quite similar to wild type with respect to cell cycle parameters. We speculate that since SeqA binding sites might overlap with DnaA binding sites spread throughout the chromosome, excess SeqA could interfere with DnaA titration and thereby increase free DnaA level. Thus, in spite of reduction in total DnaA, the amount of DnaA molecules available for initiation may not be reduced.