Enhancing plasma membrane NADPH oxidase activity increases current output by diatoms in biophotovoltaic devices

Biophotovoltaic (BPV) devices employ the photosynthetic activity of microalgae or cyanobacteria to harvest light energy and generate electrical current directly as a result of the release of electrons from the algal cells. NADPH oxidases (NOX) are plasma-membrane enzymes that transport electrons from the cytosol to generate extracellular superoxide anions, and have been implicated in BPV output. In this study, we investigated NOX activity in the diatoms, Phaeodactylum tricornutum and Thalassiosira pseudonana in an attempt to understand and enhance NOX and BPV function. We found that NOX activity was linked to defined growth regimes and growth phases, and was light dependent. Crucially, current output in a BPV device correlated with NOX activity, and levels of up to 14 μA per 106 cells (approximately 500 mA.m-2) were obtained. Expression of two putative P. tricornutum NOX genes (PtNOX1 and PtNOX2) was found to correspond with the observed growth patterns of superoxide anion production and power output, suggesting these are responsible for the observed patterns of NOX activity. Crucially, we demonstrate that NOX activity levels could be enhanced via semi-continuous culturing, pointing to the possibility of maintaining long-term power output in BPV devices.This work was supported by the United Kingdom Engineering and Physical Sciences Research Council (EPSRC), grant reference EP/F047940/1.This is the final version of the article. It first appeared from Elsevier via http://dx.doi.org/10.1016/j.algal.2015.08.00

Arabidopsis annexin1 mediates the radical-activated plasma membrane Ca2+ - and K+ -permeable conductance in root cells

Plant cell growth and stress signaling require Ca2+ influx through plasma membrane transport proteins that are regulated by reactive oxygen species. In root cell growth, adaptation to salinity stress, and stomatal closure, such proteins operate downstream of the plasma membrane NADPH oxidases that produce extracellular superoxide anion, a reactive oxygen species that is readily converted to extracellular hydrogen peroxide and hydroxyl radicals, OH_. In root cells, extracellular OH_ activates a plasma membrane Ca2+-permeable conductance that permits Ca2+ influx. In Arabidopsis thaliana, distribution of this conductance resembles that of annexin1 (ANN1). Annexins are membrane binding proteins that can form Ca2+-permeable conductances in vitro. Here, the Arabidopsis loss-of-function mutant for annexin1 (Atann1) was found to lack the root hair and epidermal OH_-activated Ca2+- and K+-permeable conductance. This manifests in both impaired root cell growth and ability to elevate root cell cytosolic free Ca2+ in response to OH_. An OH_-activated Ca2+ conductance is reconstituted by recombinant ANN1 in planar lipid bilayers. ANN1 therefore presents as a novel Ca2+-permeable transporter providing a molecular link between reactive oxygen species and cytosolic Ca2+ in plants

Adaptive mechanisms of plants against salt stress and salt shock

Salinization process occurs when soil is contaminated with salt, which consequently influences plant growth and development leading to reduction in yield of many food crops. Responding to a higher salt concentration than the normal range can result in plant developing complex physiological traits and activation of stress-related genes and metabolic pathways. Many studies have been carried out by different research groups to understand adaptive mechanism in many plant species towards salinity stress. However, different methods of sodium chloride (NaCl) applications definitely give different responses and adaptive mechanisms towards the increase in salinity. Gradual increase in NaCl application causes the plant to have salt stress or osmotic stress, while single step and high concentration of NaCl may result in salt shock or osmotic shock. Osmotic shock can cause cell plasmolysis and leakage of osmolytes in plant. Also, the gene expression pattern is influenced by the type of methods used in increasing the salinity. Therefore, this chapter discusses the adaptive mechanism in plant responding to both types of salinity increment, which include the morphological changes of plant roots and aerial parts, involvement of signalling molecules in stress perception and regulatory networks and production of osmolyte and osmoprotective proteins

Annexin 1 regulates the H(2)O(2)-induced calcium signature in Arabidopsis thaliana roots

Hydrogen peroxide is the most stable of the reactive oxygen species (ROS) and is a regulator of development, immunity and adaptation to stress. It frequently acts by elevating cytosolic free Ca2+ ([Ca2+]cyt) as a second messenger, with activation of plasma membrane Ca2+‐permeable influx channels as a fundamental part of this process. At the genetic level, to date only the Ca2+‐permeable Stelar K+ Outward Rectifier (SKOR) channel has been identified as being responsive to hydrogen peroxide. We show here that the ROS‐regulated Ca2+ transport protein Annexin 1 in Arabidopsis thaliana (AtANN1) is involved in regulating the root epidermal [Ca2+]cyt response to stress levels of extracellular hydrogen peroxide. Peroxide‐stimulated [Ca2+]cyt elevation (determined using aequorin luminometry) was aberrant in roots and root epidermal protoplasts of the Atann1 knockout mutant. Similarly, peroxide‐stimulated net Ca2+ influx and K+ efflux were aberrant in Atann1 root mature epidermis, determined using extracellular vibrating ion‐selective microelectrodes. Peroxide induction of GSTU1 (Glutathione‐S‐Transferase1 Tau 1), which is known to be [Ca2+]cyt‐dependent was impaired in mutant roots, consistent with a lesion in signalling. Expression of AtANN1 in roots was suppressed by peroxide, consistent with the need to restrict further Ca2+ influx. Differential regulation of annexin expression was evident, with AtANN2 down‐regulation but up‐regulation of AtANN3 and AtANN4. Overall the results point to involvement of AtANN1 in shaping the root peroxide‐induced [Ca2+]cyt signature and downstream signalling.Siân L. Richards, Anuphon Laohavisit, Jennifer C. Mortimer, Lana Shabala Stéphanie M. Swarbreck, Sergey Shabala, Julia M. Davie

Annexin-mediated calcium signalling in roots

Annexin-mediated calcium signalling in roots. XVI International Workshop on Plant Membrane Biolog

<i>Zea mays</i> Annexins Modulate Cytosolic Free Ca2+ and Generate a Ca2+-Permeable Conductance

Abstract Regulation of reactive oxygen species and cytosolic free calcium ([Ca2+]cyt) is central to plant function. Annexins are small proteins capable of Ca2+-dependent membrane binding or membrane insertion. They possess structural motifs that could support both peroxidase activity and calcium transport. Here, a Zea mays annexin preparation caused increases in [Ca2+]cyt when added to protoplasts of Arabidopsis thaliana roots expressing aequorin. The pharmacological profile was consistent with annexin activation (at the extracellular plasma membrane face) of Arabidopsis Ca2+-permeable nonselective cation channels. Secreted annexins could therefore modulate Ca2+ influx. As maize annexins occur in the cytosol and plasma membrane, they were incorporated at the intracellular face of lipid bilayers designed to mimic the plasma membrane. Here, they generated an instantaneously activating Ca2+-permeable conductance at mildly acidic pH that was sensitive to verapamil and Gd3+ and had a Ca2+-to-K+ permeability ratio of 0.36. These results suggest that cytosolic annexins create a Ca2+ influx pathway directly, particularly during stress responses involving acidosis. A maize annexin preparation also demonstrated in vitro peroxidase activity that appeared independent of heme association. In conclusion, this study has demonstrated that plant annexins create Ca2+-permeable transport pathways, regulate [Ca2+]cyt, and may function as peroxidases in vitro.</jats:p

ANXUR receptor-like kinases coordinate cell wall integrity with growth at the pollen tube tip via NADPH oxidases

It has become increasingly apparent that the extracellular matrix (ECM), which in plants corresponds to the cell wall, can influence intracellular activities in ways that go far beyond their supposedly passive mechanical support. In plants, growing cells use mechanisms sensing cell wall integrity to coordinate cell wall performance with the internal growth machinery to avoid growth cessation or loss of integrity. How this coordination precisely works is unknown. Previously, we reported that in the tip-growing pollen tube the ANXUR receptor-like kinases (RLKs) of the CrRLK1L subfamily are essential to sustain growth without loss of cell wall integrity in Arabidopsis. Here, we show that over-expression of the ANXUR RLKs inhibits growth by over-activating exocytosis and the over-accumulation of secreted cell wall material. Moreover, the characterization of mutations in two partially redundant pollen-expressed NADPH oxidases coupled with genetic interaction studies demonstrate that the ANXUR RLKs function upstream of these NADPH oxidases. Using the H₂O₂-sensitive HyPer and the Ca²⁺-sensitive YC3.60 sensors in NADPH oxidase-deficient mutants, we reveal that NADPH oxidases generate tip-localized, pulsating H₂O₂ production that functions, possibly through Ca²⁺ channel activation, to maintain a steady tip-focused Ca²⁺ gradient during growth. Our findings support a model where ECM-sensing receptors regulate reactive oxygen species production, Ca²⁺ homeostasis, and exocytosis to coordinate ECM-performance with the internal growth machinery