Local measurements of the mean interstellar polarization at high Galactic latitudes

Very little information exists concerning the properties of the interstellar medium (ISM)-induced starlight polarization at high Galactic latitudes. Future optopolarimetric surveys promise to fill this gap. We conduct a small-scale pathfinding survey designed to identify the average polarization properties of the diffuse ISM locally, at regions with the lowest dust content. We perform deep optopolarimetric surveys within three ~15′× 15′ regions located at b > 48° using the RoboPol polarimeter. The observed samples of stars are photometrically complete to ~16 mag in the R-band. The selected regions exhibit low total reddening compared to the majority of high-latitude sightlines. We measure the level of systematic uncertainty for all observing epochs and find it to be 0.1% in fractional linear polarization, p. The majority of individual stellar measurements have low signal-to-noise ratios. However, our survey strategy enables us to locate the mean fractional linear polarization p_(mean) in each of the three regions. The region with lowest dust content yields p_(mean) = (0.054 ± 0.038)%, not significantly different from zero. We find significant detections for the remaining two regions of: p_(mean) = (0.113 ± 0.036)% and p_(mean) = (0.208 ± 0.044)%. Using a Bayesian approach, we provide upper limits on the intrinsic spread of the small-scale distributions of q and u. At the detected p_(mean) levels, the determination of the systematic uncertainty is critical for the reliability of the measurements. We verify the significance of our detections with statistical tests, accounting for all sources of uncertainty. Using publicly available HI emission data, we identify the velocity components that most likely account for the observed p_(mean) and find their morphologies to be misaligned with the orientation of the mean polarization at a spatial resolution of 10′. We find indications that the standard upper envelope of p with reddening underestimates the maximum p at very low E(B–V) (≤0.01 mag)

Immunolocalization of dually phosphorylated MAPKs in dividing root meristem cells of Vicia faba, Pisum sativum, Lupinus luteus and Lycopersicon esculentum

Key message In plants, phosphorylated MAPKs display constitutive nuclear localization; however, not all studied plant species show co-localization of activated MAPKs to mitotic microtubules. Abstract The mitogen-activated protein kinase (MAPK) signaling pathway is involved not only in the cellular response to biotic and abiotic stress but also in the regulation of cell cycle and plant development. The role of MAPKs in the formation of a mitotic spindle has been widely studied and the MAPK signaling pathway was found to be indispensable for the unperturbed course of cell division. Here we show cellular localization of activated MAPKs (dually phosphorylated at their TXY motifs) in both interphase and mitotic root meristem cells of Lupinus luteus, Pisum sativum, Vicia faba (Fabaceae) and Lycopersicon esculentum (Solanaceae). Nuclear localization of activated MAPKs has been found in all species. Colocalization of these kinases to mitotic microtubules was most evident in L. esculentum, while only about 50 % of mitotic cells in the root meristems of P. sativum and V. faba displayed activated MAPKs localized to microtubules during mitosis. Unexpectedly, no evident immunofluorescence signals at spindle microtubules and phragmoplast were noted in L. luteus. Considering immunocytochemical analyses and studies on the impact of FR180204 (an inhibitor of animal ERK1/2) on mitotic cells, we hypothesize that MAPKs may not play prominent role in the regulation of microtubule dynamics in all plant species

Strategic and practical guidelines for successful structured illumination microscopy

Linear 2D- or 3D-structured illumination microscopy (SIM or3D-SIM, respectively) enables multicolor volumetric imaging of fixed and live specimens with subdiffraction resolution in all spatial dimensions. However, the reliance of SIM on algorithmic post-processing renders it particularly sensitive to artifacts that may reduce resolution, compromise data and its interpretations, and drain resources in terms of money and time spent. Here we present a protocol that allows users to generate high-quality SIM data while accounting and correcting for common artifacts. The protocol details preparation of calibration bead slides designed for SIM-based experiments, the acquisition of calibration data, the documentation of typically encountered SIM artifacts and corrective measures that should be taken to reduce them. It also includes a conceptual overview and checklist for experimental design and calibration decisions, and is applicable to any commercially available or custom platform. This protocol, plus accompanying guidelines, allows researchers from students to imaging professionals to create an optimal SIM imaging environment regardless of specimen type or structure of interest. The calibration sample preparation and system calibration protocol can be executed within 1-2 d

A Bacterial Acetyltransferase Destroys Plant Microtubule Networks and Blocks Secretion

The eukaryotic cytoskeleton is essential for structural support and intracellular transport, and is therefore a common target of animal pathogens. However, no phytopathogenic effector has yet been demonstrated to specifically target the plant cytoskeleton. Here we show that the Pseudomonas syringae type III secreted effector HopZ1a interacts with tubulin and polymerized microtubules. We demonstrate that HopZ1a is an acetyltransferase activated by the eukaryotic co-factor phytic acid. Activated HopZ1a acetylates itself and tubulin. The conserved autoacetylation site of the YopJ / HopZ superfamily, K289, plays a critical role in both the avirulence and virulence function of HopZ1a. Furthermore, HopZ1a requires its acetyltransferase activity to cause a dramatic decrease in Arabidopsis thaliana microtubule networks, disrupt the plant secretory pathway and suppress cell wall-mediated defense. Together, this study supports the hypothesis that HopZ1a promotes virulence through cytoskeletal and secretory disruption

Hyperosmotic stress induces formation of tubulin macrotubules in root-tip cells of Triticum turgidum: Their probable involvement in protoplast volume control

Treatment of root-tip cells of Triticum turgidum with 1 M mannitol solution for 30 min induces microtubule (Mt) disintegration in the plasmolyzed protoplasts. Interphase plasmolyzed cells possess many cortical, perinuclear and endoplasmic macrotubules, 35 nm in mean diameter, forming prominent arrays. In dividing cells macrotubules assemble into aberrant mitotic and cytokinetic apparatuses resulting in the disturbance of cell division. Putative tubulin paracrystals were occasionally observed in plasmolyzed cells. The quantity of polymeric tubulin in plasmolyzed cells exceeds that in control cells. Root-tip cells exposed for 2-8 h to plasmolyticum recover partially, although the volume of the plasmolyzed protoplast does not change detectably. Among other events, the macrotubules are replaced by Mts, chromatin assumes its typical appearance and the cells undergo typical cell divisions. Additionally, polysaccharidic material is found in the periplasmic space. Oryzalin and colchicine treatment induced macrotubule disintegration and a significant reduction of protoplast volume in every plasmolyzed cell type examined, whereas cytochalasin B had only minor effects restricted to differentiated cells. These results suggest that Mt destruction by hyperosmotic stress, and their replacement by tubulin macrotubules and putative tubulin paracrystals is a common feature among angiosperms and that macrotubules are involved in the mechanism of protoplast volume regulation

Actomyosin is involved in the plasmolytic cycle: Gliding movement of the deplasmolyzing protoplast

The leaf cells of Chlorophytum comosum seem to have the ability to regulate their protoplast volume and shape during the plasmolytic cycle. This phenomenon was morphologically expressed by the stabilization of the plasmolyzed protoplast volume and shape within 1-5 min after the immersion of the leaf segments in the plasmolytic fluid and temporarily at the onset of deplasmolysis. During the latter stage the plasmolyzed protoplast rounded up and assumed a perfectly convex shape and glided into the cell lumen along the cell axis. This gliding movement was active, nonsaltatory, and conducted with a constant velocity and lasted for a short time. During this movement the protoplast volume did not change appreciably. As far as we know, this movement has not been described so far. Deplasmolysis proceeded and was rapidly completed when the protoplast stopped moving. Leaf cells which have been affected by an antiactin filament drug or myosin inhibitors lost their ability to regulate the volume and shape of the plasmolyzing protoplast. In addition, the gliding protoplast movement was also inhibited in the treated cells. These data show for the first time that the actomyosin system is involved in the mechanism of volume regulation during the plasmolytic cycle and that it underlies the gliding movement of the deplasmolyzing protoplast

Actomyosin is involved in the plasmolytic cycle: Gliding movement of the deplasmolyzing protoplast

The leaf cells of Chlorophytum comosum seem to have the ability to regulate their protoplast volume and shape during the plasmolytic cycle. This phenomenon was morphologically expressed by the stabilization of the plasmolyzed protoplast volume and shape within 1-5 min after the immersion of the leaf segments in the plasmolytic fluid and temporarily at the onset of deplasmolysis. During the latter stage the plasmolyzed protoplast rounded up and assumed a perfectly convex shape and glided into the cell lumen along the cell axis. This gliding movement was active, nonsaltatory, and conducted with a constant velocity and lasted for a short time. During this movement the protoplast volume did not change appreciably. As far as we know, this movement has not been described so far. Deplasmolysis proceeded and was rapidly completed when the protoplast stopped moving. Leaf cells which have been affected by an antiactin filament drug or myosin inhibitors lost their ability to regulate the volume and shape of the plasmolyzing protoplast. In addition, the gliding protoplast movement was also inhibited in the treated cells. These data show for the first time that the actomyosin system is involved in the mechanism of volume regulation during the plasmolytic cycle and that it underlies the gliding movement of the deplasmolyzing protoplast