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Colin Brownlee

Publications and source records attributed to Colin Brownlee.

13 recordsLinked to original sources
PubMed2006

Acyl-homoserine lactones modulate the settlement rate of zoospores of the marine alga Ulva intestinalis via a novel chemokinetic mechanism.

Bacteria utilize quorum sensing to regulate the expression of cell density-dependant phenotypes such as biofilm formation and virulence. Zoospores of the marine alga Ulva intestinalis exploit the acyl-homoserine lactone (AHL) quorum sensing system to identify bacterial biofilms for preferential settlement. Here, we demonstrate that AHLs act as strong chemoattractants for Ulva zoospores. Chemoattraction does not involve a chemotactic orientation towards the AHL source. Instead, it occurs through a chemokinesis in which zoospore swimming speed is rapidly decreased in the presence of AHLs. The chemoresponse to AHLs was dependant on the nature of the acyl side chain, with N-(3-oxododecanoyl)-homoserine lactone (30-C12-HSL) being the most effective signal molecule. Mean zoospore swimming speed decreased more rapidly over wild-type biofilms of the marine bacteria Vibrio anguillarum relative to biofilms of the vanM mutant, in which AHL synthesis is disrupted. These data implicate a role for AHL-mediated chemokinesis in the location and preferential settlement of Ulva zoospores on marine bacterial assemblages. Exposure to AHLs did not inhibit the negative phototaxis of Ulva zoospores, indicating that chemoattraction to bacterial biofilms does not preclude the response to a light stimulus in substrate location.

4-Butyrolactone↗
PubMed2006

Biolistic delivery of Ca2+ dyes into plant and algal cells.

In eukaryotes, changes in cytosolic Ca2+ concentrations ([Ca2+]cyt) are associated with a number of environmental and developmental stimuli. However, measuring [Ca2+]cyt changes in single plant or algal cells is often problematic. Although a wide range of Ca2+-sensitive fluorescent dyes is available, they are often difficult to introduce into plant cells. Micro-injection is the most robust method for dye loading, but is time-consuming, technically demanding, and unsuitable in many cell types. To overcome these problems, we have adapted biolistic techniques to load Ca2+-sensitive dyes into guard cells of the flowering plant, Commelina communis, cells of the green alga Chlamydomonas reinhardtii, and zygotes of the brown alga, Fucus serratus. Using this approach, we have been able to monitor [Ca2+]cyt changes in response to various stimuli, including a novel [Ca2+]cyt response in C. reinhardtii. The method allows the use of free acid and dextran-conjugated dyes. Biolistic loading of differentiated plant cells is easier, quicker, and more widely applicable than micro-injection, and should broaden the study of plant signal transduction.

Animals↗
PubMed2005

Prior exposure to Cu contamination influences the outcome of toxicological testing of Fucus serratus embryos.

Brown seaweeds are often employed in single species toxicity testing to study the association between the pollutant and the biota in contaminated marine habitats. We have used Fucus serratus (Phaeophyta) from one Cu resistant and one non-resistant population to evaluate the effect of prior exposure to metal pollution on toxicological endpoints. Analysis of comparative toxicity was conducted for embryo rhizoid elongation and adult relative growth rate (RGR). Algae that had previously been exposed to Cu expressed consistently lower levels of sensitivity to Cu than those that had no history of exposure to the pollutant. For both non-resistant and resistant populations rhizoid length was a more sensitive endpoint than adult RGR. While early life history stages of brown algae are generally regarded as being pollution-sensitive and inhibition of spore and embryo rhizoid elongation is frequently used as endpoints in bioassays, the test results may be affected by prior exposure of the parent algae to the pollutant. We conclude that the effect of prior exposure should be considered when comparing endpoints between studies and when selecting material for future testing.

Bias↗
PubMed2005

Spatial re-organisation of cortical microtubules in vivo during polarisation and asymmetric division of Fucus zygotes.

Fucus zygotes polarise and germinate a rhizoid before their first asymmetrical division. The role of microtubules (MTs) in orienting the first division plane has been extensively studied by immunofluorescence approaches. In the present study, the re-organisation of MT arrays during the development of Fucus zygotes and embryos was followed in vivo after microinjection of fluorescent tubulin. A dynamic cortical MT array that shows dramatic reorganization during zygote polarization was detected for the first time. Randomly distributed cortical MTs were redistributed to the presumptive rhizoid site by the time of polarisation and well before rhizoid germination. The cortical MT re-organisation occurs independently of centrosome separation and nucleation. By the time of mitosis the cortical array depolymerised to cortical foci in regions from which it also reformed following mitosis, suggesting that it is nucleated from cortical sites. We confirm previous indications from immunodetection studies that centrosomal alignment and nuclear rotation occur via MT connexions to stabilised cortical sites and that definitive alignment is post-metaphasic. Finally, we show that cortical MTs align parallel to the growth axis during rhizoid tip growth and our results suggest that they may be involved in regulating rhizoid growth by shaping the rhizoid and containing turgor pressure.

Antineoplastic Agents↗
PubMed2004

The generation of Ca(2+) signals in plants.

The calcium ion is firmly established as a ubiquitous intracellular second messenger in plants. At their simplest, Ca(2+)-based signaling systems are composed of a receptor, a system for generating the increase in [Ca(2+)]cyt, downstream components that are capable of reacting to the increase in [Ca(2+)]cyt, and other cellular systems responsible for returning [Ca(2+)]cyt to its prestimulus level. Here we review the various mechanisms responsible for generating the stimulus-induced increases in [Ca(2+)]cyt known as Ca(2+) signals. We focus particularly on the mechanisms responsible for generating [Ca(2+)]cyt oscillations and transients and use Nod Factor signaling in legume root hairs and stimulus-response coupling in guard cells to assess the physiological significance of these classes of Ca(2+) signals.

Calcium↗
PubMed2003

Inositol hexakisphosphate mobilizes an endomembrane store of calcium in guard cells.

myo-Inositol hexakisphosphate (InsP6) is the most abundant inositol phosphate in cells, yet it remains the most enigmatic of this class of signaling molecule. InsP6 plays a role in the processes by which the drought stress hormone abscisic acid (ABA) induces stomatal closure, conserving water and ensuring plant survival. Previous work has shown that InsP6 levels in guard cells are elevated in response to ABA, and InsP6 inactivates the plasma membrane inward K+ conductance (IK,in) in a cytosolic calcium-dependent manner. The use of laser-scanning confocal microscopy in dye-loaded patch-clamped guard cell protoplasts shows that release of InsP6 from a caged precursor mobilizes calcium. Measurement of calcium (barium) currents ICa in patch-clamped protoplasts in whole cell mode shows that InsP6 has no effect on the calcium-permeable channels in the plasma membrane activated by ABA. The InsP6-mediated inhibition of IK,in can also be observed in the absence of external calcium. Thus the InsP6-induced increase in cytoplasmic calcium does not result from calcium influx but must arise from InsP6-triggered release of calcium from endomembrane stores. Measurements of vacuolar currents in patch-clamped isolated vacuoles in whole-vacuole mode showed that InsP6 activates both the fast and slow conductances of the guard cell vacuole. These data define InsP6 as an endomembrane-acting calcium-release signal in guard cells; the vacuole may contribute to InsP6-triggered Ca2+ release, but other endomembranes may also be involved.

Abscisic Acid↗
PubMed2003

Reactive oxygen species produced by NADPH oxidase regulate plant cell growth.

Cell expansion is a central process in plant morphogenesis, and the elongation of roots and root hairs is essential for uptake of minerals and water from the soil. Ca2+ influx from the extracellular store is required for (and sets the rates of) cell elongation in roots. Arabidopsis thaliana rhd2 mutants are defective in Ca2+ uptake and consequently cell expansion is compromised--rhd2 mutants have short root hairs and stunted roots. To determine the regulation of Ca2+ acquisition in growing root cells we show here that RHD2 is an NADPH oxidase, a protein that transfers electrons from NADPH to an electron acceptor leading to the formation of reactive oxygen species (ROS). We show that ROS accumulate in growing wild-type (WT) root hairs but their levels are markedly decreased in rhd2 mutants. Blocking the activity of the NADPH oxidase with diphenylene iodonium (DPI) inhibits ROS formation and phenocopies Rhd2-. Treatment of rhd2 roots with ROS partly suppresses the mutant phenotype and stimulates the activity of plasma membrane hyperpolarization-activated Ca2+ channels, the predominant root Ca2+ acquisition system. This indicates that NADPH oxidases control development by making ROS that regulate plant cell expansion through the activation of Ca2+ channels.

Alleles↗
PubMed2003

A novel Cl- inward-rectifying current in the plasma membrane of the calcifying marine phytoplankton Coccolithus pelagicus.

We investigated the membrane properties and dominant ionic conductances in the plasma membrane of the calcifying marine phytoplankton Coccolithus pelagicus using the patch-clamp technique. Whole-cell recordings obtained from decalcified cells revealed a dominant anion conductance in response to membrane hyperpolarization. Ion substitution showed that the anion channels were selective for Cl(-) and Br(-) over other anions, and the sensitivity to the stilbene derivative 4,4'-diisothiocyanatostilbene-2,2'-disulfonic acid, ethacrynic acid, and Zn(2+) revealed a pharmacological profile typical of many plant and animal anion channels. Voltage activation and kinetic characteristics of the C. pelagicus Cl(-) channel are consistent with a novel function in plants as the inward rectifier that tightly regulates membrane potential. Membrane depolarization gave rise to nonselective cation currents and in some cases evoked action potential currents. We propose that these major ion conductances play an essential role in membrane voltage regulation that relates to the unique transport physiology of these calcifying phytoplankton.

Biological Transport↗
PubMed2002

Plant K+ transport: not just an uphill struggle.

Most plant cells can accumulate K+ to concentrations much higher than those in the external medium. Recent studies are providing new insights into the relative roles of channels and transporters in K+ accumulation. These studies are also pointing to specific roles for K+ channels and transporters in polarized development.

Ion Transport↗
PubMed2002

Role of the extracellular matrix in cell-cell signalling: paracrine paradigms.

The plant extracellular matrix (ECM) is complex and diverse, and is involved in cell-cell communication in a wide range of developmental, reproductive and pathogenic processes. Characterisation of integral ECM components is leading to improved understanding of their roles in signalling. Interactions between the extracellular domains of plant plasma membrane receptor kinases and their ligands are potentially regulated by the properties of the ECM. Several of these interactions, for example those involving the S-locus receptor kinase, are being characterised in some detail. Non-protein constituents are also implicated in regulating the movement of signalling molecules in the ECM, which is associated with developmental patterning. In contrast to the situation in animal cells, cytoskeleton-integrin-ECM signalling complexes appear not to be dominant features of signal transduction in plant cells. Nevertheless, structural adhesions between the plasma membrane and cell wall are important for a variety of functions.

Cell Differentiation↗
PubMed2002

Spatiotemporal patterning of reactive oxygen production and Ca(2+) wave propagation in fucus rhizoid cells.

Both Ca(2+) and reactive oxygen species (ROS) play critical signaling roles in plant responses to biotic and abiotic stress. However, the positioning of Ca(2+) and ROS (in particular H(2)O(2)) after a stress stimulus and their subcellular interactions are poorly understood. Moreover, although information can be encoded in different patterns of cellular Ca(2+) signals, little is known about the subcellular spatiotemporal patterns of ROS production or their significance for downstream responses. Here, we show that ROS production in response to hyperosmotic stress in embryonic cells of the alga Fucus serratus consists of two distinct components. The first ROS component coincides closely with the origin of a Ca(2+) wave in the peripheral cytosol at the growing cell apex, has an extracellular origin, and is necessary for the Ca(2+) wave. Patch-clamp experiments show that a nonselective cation channel is stimulated by H(2)O(2) and may underlie the initial cytosolic Ca(2+) increase. Thus, the spatiotemporal pattern of the Ca(2+) wave is determined by peripheral ROS production. The second, later ROS component localizes to the mitochondria and is a direct consequence of the Ca(2+) wave. The first component, but not the second, is required for short-term adaptation to hyperosmotic stress. Our results highlight the role of ROS in the patterning of a Ca(2+) signal in addition to its function in regulating cell wall strength in the Fucus embryo.

Adaptation, Physiological↗