Browsing by Subject "NADPH OXIDASE RBOHD"

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  • Deger, Aysin Guzel; Scherzer, Sönke; Nuhkat, Maris; Kedzierska, Justyna; Kollist, Hannes; Brosche, Mikael; Unyayar, Serpil; Boudsocq, Marie; Hedrich, Rainer; Roelfsema, M. Rob G. (2015)
    During infection plants recognize microbe-associated molecular patterns (MAMPs), and this leads to stomatal closure. This study analyzes the molecular mechanisms underlying this MAMP response and its interrelation with ABA signaling. Stomata in intact Arabidopsis thaliana plants were stimulated with the bacterial MAMP flg22, or the stress hormone ABA, by using the noninvasive nanoinfusion technique. Intracellular double-barreled microelectrodes were applied to measure the activity of plasma membrane ion channels. Flg22 induced rapid stomatal closure and stimulated the SLAC1 and SLAH3 anion channels in guard cells. Loss of both channels resulted in cells that lacked flg22-induced anion channel activity and stomata that did not close in response to flg22 or ABA. Rapid flg22-dependent stomatal closure was impaired in plants that were flagellin receptor (FLS2)-deficient, as well as in the ost1-2 (Open Stomata 1) mutant, which lacks a key ABA-signaling protein kinase. By contrast, stomata of the ABA protein phosphatase mutant abi1-1 (ABscisic acid Insensitive 1) remained flg22-responsive. These data suggest that the initial steps in flg22 and ABA signaling are different, but that the pathways merge at the level of OST1 and lead to activation of SLAC1 and SLAH3 anion channels.
  • Overmyer, Kirk Loren; Vuorinen, Katariina Elina; Brosche, Mikael Johan (2018)
    Plants live in a world where they are challenged by abiotic and biotic stresses. In response to unfavorable conditions or an acute challenge like a pathogen attack, plants use various signaling pathways that regulate expression of defense genes and other mechanisms to provide resistance or stress adaptation. Identification of the regulatory steps in defense signaling has seen much progress in recent years. Many of the identified signaling pathways show interactions with each other, exemplified by the modulation of the jasmonic acid response by salicylic acid. Accordingly, defense regulation is more appropriately thought of as a web of interactions, rather than linear pathways. Here we describe various regulatory components and how they interact to provide an appropriate defense response. One of the common assays to monitor the output of defense signaling, as well as interaction between signaling pathways, is the measurement of altered gene expression. We illustrate that, while this is a suitable assay to monitor defense regulation, it can also inadvertently provide overstated conclusions about interaction among signaling pathways.
  • Durian, Guido; Rahikainen, Moona; Alegre, Sara; Brosche, Mikael; Kangasjarvi, Saijaliisa (2016)
    Biotic stress factors pose a major threat to plant health and can significantly deteriorate plant productivity by impairing the physiological functions of the plant. To combat the wide range of pathogens and insect herbivores, plants deploy converging signaling pathways, where counteracting activities of protein kinases and phosphatases form a basic mechanism for determining appropriate defensive measures. Recent studies have identified Protein Phosphatase 2A (PP2A) as a crucial component that controls pathogenesis responses in various plant species. Genetic, proteomic and metabolomic approaches have underscored the versatile nature of PP2A, which contributes to the regulation of receptor signaling, organellar signaling, gene expression, metabolic pathways, and cell death, all of which essentially impact plant immunity. Associated with this, various PP2A subunits mediate post-translational regulation of metabolic enzymes and signaling components. Here we provide an overview of protein kinase/phosphatase functions in plant immunity signaling, and position the multifaceted functions of PP2A in the tightly inter-connected regulatory network that controls the perception, signaling and responding to biotic stress agents in plants.
  • Ehonen, Sanna; Yarmolinsky, Dmitry; Kollist, Hannes; Kangasjärvi, Jaakko (2019)
    Significance: Stomata sense the intercellular carbon dioxide (CO2) concentration (C-i) and water availability under changing environmental conditions and adjust their apertures to maintain optimal cellular conditions for photosynthesis. Stomatal movements are regulated by a complex network of signaling cascades where reactive oxygen species (ROS) play a key role as signaling molecules. Recent Advances: Recent research has uncovered several new signaling components involved in CO2- and abscisic acid-triggered guard cell signaling pathways. In addition, we are beginning to understand the complex interactions between different signaling pathways. Critical Issues: Plants close their stomata in reaction to stress conditions, such as drought, and the subsequent decrease in C-i leads to ROS production through photorespiration and over-reduction of the chloroplast electron transport chain. This reduces plant growth and thus drought may cause severe yield losses for agriculture especially in arid areas. Future Directions: The focus of future research should be drawn toward understanding the interplay between various signaling pathways and how ROS, redox, and hormonal balance changes in space and time. Translating this knowledge from model species to crop plants will help in the development of new drought-resistant crop species with high yields. Antioxid. Redox Signal. 00, 000-000.
  • Nickolov, Kaloian; Gauthier, Adrien; Hashimoto, Kenji; Laitinen, Teresa; Väisänen, Enni; Paasela, Tanja; Soliymani, Rabah; Kurusu, Takamitsu; Himanen, Kristiina; Blokhina, Olga; Fagerstedt, Kurt; Jokipii-Lukkari, Soile; Tuominen, Hannele; Haggman, Hely; Wingsle, Gunnar; Teeri, Teemu H.; Kuchitsu, Kazuyuki; Kärkönen, Anna (2022)
    Plant respiratory burst oxidase homologs (RBOHs) are plasma membrane-localized NADPH oxidases that generate superoxide anion radicals, which then dismutate to H2O2, into the apoplast using cytoplasmic NADPH as an electron donor. PaRBOH1 is the most highly expressed RBOH gene in developing xylem as well as in a lignin-forming cell culture of Norway spruce (Picea abies L. Karst.). Since no previous information about regulation of gymnosperm RBOHs exist, our aim was to resolve how PaRBOH1 is regulated with a focus on phosphorylation. The N-terminal part of PaRBOH1 was found to contain several putative phosphorylation sites and a four-times repeated motif with similarities to the Botrytis-induced kinase 1 target site in Arabidopsis AtRBOHD. Phosphorylation was indicated for six of the sites in in vitro kinase assays using 15 amino-acid-long peptides for each of the predicted phosphotarget site in the presence of protein extracts of developing xylem. Serine and threonine residues showing positive response in the peptide assays were individually mutated to alanine (kinase-inactive) or to aspartate (phosphomimic), and the wild type PaRBOH1 and the mutated constructs transfected to human kidney embryogenic (HEK293T) cells with a low endogenous level of extracellular ROS production. ROS-producing assays with HEK cells showed that Ca2+ and phosphorylation synergistically activate the enzyme and identified several serine and threonine residues that are likely to be phosphorylated including a novel phosphorylation site not characterized in other plant species. These were further investigated with a phosphoproteomic study. Results of Norway spruce, the first gymnosperm species studied in relation to RBOH regulation, show that regulation of RBOH activity is conserved among seed plants.