scholarly journals Abscisic Acid Increases Hydrogen Peroxide in Multiple Subcellular Compartments to Drive Stomatal Closure

2022 ◽  
Author(s):  
Anthony E Postliglione ◽  
Gloria K Muday
Keyword(s):  
Hydrogen Peroxide ◽  
Stomatal Closure ◽  
Subcellular Location ◽  
Guard Cells ◽  
Subcellular Compartments ◽  
Hydrogen Peroxide Treatment ◽  
Genes Encoding ◽  
Aba Insensitive ◽  
Necessary And Sufficient ◽  
Respiratory Burst Oxidase

Stomatal closure regulates transpiration and gas exchange in response to environmental cues. Drought upregulates ABA signaling, which elevates levels of reactive oxygen species (ROS). However, the subcellular location and identity of these ROS has received limited study. We found that in guard cells, ABA increased fluorescence of the general redox sensor, dichlorofluorescein (DCF), in distinct subcellular locations including chloroplasts, cytosol, nuclei, and cytosolic puncta. These changes were lost in ABA-insensitive quintuple receptor mutant and accentuated in an ABA-hypersensitive mutant. ABA induced ROS accumulation in these subcellular compartments was lost in mutants with defects in genes encoding hydrogen peroxide synthesizing respiratory burst oxidase homolog (RBOH) enzymes and guard cells treated with the RBOH inhibitor VAS2870, while exogenous hydrogen peroxide treatment is sufficient to close guard cells. The hydrogen peroxide-selective probe, peroxy orange1, also showed ABA-dependent increases in chloroplasts and cytosolic puncta. Using the more sensitive genetically-encoded hydrogen peroxide reporter roGFP-Orp1, we also detected significant hydrogen peroxide increases in the cytosol and nucleus. These cytosolic puncta accumulate ROS after ABA treatment show colocalization with Mitotracker and with a mitochondrial targeted mt-roGFP2-Orp1, which also revealed ABA-increased ROS in mitochondria. These results indicate that elevated hydrogen peroxide after ABA treatment in these subcellular compartments is necessary and sufficient to drive stomatal closure.

Hydrogen peroxide is a common signal for darkness- and ABA-induced stomatal closure in Pisum sativum

2004 ◽  
Vol 31 (9) ◽  
pp. 913 ◽  
Author(s):  
Radhika Desikan ◽  
Man-Kim Cheung ◽  
Andrew Clarke ◽  
Sarah Golding ◽  
Moshe Sagi ◽  
...  
Keyword(s):  
Hydrogen Peroxide ◽  
Pisum Sativum ◽  
Nadph Oxidase ◽  
Stomatal Closure ◽  
Guard Cells ◽  
Dependent Manner ◽  
Degenerate Oligonucleotide ◽  
Diphenylene Iodonium ◽  
Common Signal ◽  
Respiratory Burst Oxidase

The requirement for hydrogen peroxide (H2O2) generation and action during stomatal closure induced by darkness and abscisic acid (ABA) was investigated in pea (Pisum sativum L.). Stomatal closure induced by darkness or ABA was inhibited by the H2O2-scavenging enzyme catalase or the antioxidant N-acetyl cysteine (NAC), or by diphenylene iodonium (DPI), an inhibitor of the H2O2-generating enzyme NADPH oxidase. Exogenous H2O2 induced stomatal closure in a dose- and time-dependent manner, and H2O2 was also required for ABA-inhibition of stomatal opening in the light. H2O2 accumulation in guard cells was increased by darkness or ABA, as assessed with the fluorescent dye dichlorodihydrofluorescein diacetate (H2-DCFDA) and confocal microscopy. Such increases were inhibited by catalase, NAC or DPI, consistent with the effects of these compounds on stomatal apertures. Employing polymerase chain reaction (PCR) with degenerate oligonucleotide primers, several NADPH oxidase homologues were identified from pea genomic DNA that had substantial identity to the Arabidopsis thaliana (L.) Heynh. rboh (respiratory burst oxidase homologue) genes. Furthermore, an antibody raised against the tomato rboh identified immunoreactive proteins in epidermal, mesophyll and guard cells.

scholarly journals Dual Role of Hydrogen Peroxide in Arabidopsis Guard Cells in Response to Sulfur Dioxide

2014 ◽  
Vol 2014 ◽  
pp. 1-9 ◽  
Author(s):  
Huilan Yi ◽  
Xin Liu ◽  
Min Yi ◽  
Gang Chen
Keyword(s):  
Hydrogen Peroxide ◽  
Cell Death ◽  
Sulfur Dioxide ◽  
Nadph Oxidase ◽  
Stomatal Closure ◽  
Critical Role ◽  
Guard Cells ◽  
Oxidase Inhibitor ◽  
Air Pollutant ◽  
Low Levels

Sulfur dioxide (SO2) is a major air pollutant and has significant impacts on plant physiology. Plant can adapt to SO2 stress by controlling stomatal movement, gene expression, and metabolic changes. Here we show clear evidences that SO2-triggered hydrogen peroxide (H2O2) production mediated stomatal closure and cell death in Arabidopsis leaves. High levels of SO2 caused irreversible stomatal closure and decline in guard cell viability, but low levels of SO2 caused reversible stomatal closure. Exogenous antioxidants ascorbic acid (AsA) and catalase (CAT) or Ca2+ antagonists EGTA and LaCl3 blocked SO2-induced stomatal closure and decline in viability. AsA and CAT also blocked SO2-induced H2O2 and [Ca2+]cyt elevation. However, EGTA and LaCl3 inhibited SO2-induced [Ca2+]cyt increase but did not suppress SO2-induced H2O2 elevation. These results indicate that H2O2 elevation triggered stomatal closure and cell death via [Ca2+]cyt signaling in SO2-stimulated Arabidopsis guard cells. NADPH oxidase inhibitor DPI blocked SO2-induced cell death but not the stomatal closure triggered by low levels of SO2, indicating that NADPH oxidase-dependent H2O2 production plays critical role in SO2 toxicity but is not necessary for SO2-induced stomatal closure. Our results suggest that H2O2 production and accumulation in SO2-stimulated plants trigger plant adaptation and toxicity via reactive oxygen species mediating Ca2+ signaling.

Hydrogen sulfide may function downstream of hydrogen peroxide in salt stress-induced stomatal closure in Vicia faba

2019 ◽  
Vol 46 (2) ◽  
pp. 136 ◽  
Author(s):  
Yinli Ma ◽  
Wei Zhang ◽  
Jiao Niu ◽  
Yu Ren ◽  
Fan Zhang
Keyword(s):  
Hydrogen Peroxide ◽  
Hydrogen Sulfide ◽  
Salt Stress ◽  
Vicia Faba ◽  
Laser Scanning ◽  
Stomatal Closure ◽  
Guard Cells ◽  
H2o2 Production ◽  
Diphenylene Iodonium ◽  
Cysteine Desulfhydrase

The roles of hydrogen sulfide (H2S) and hydrogen peroxide (H2O2) in signalling transduction of stomatal closure induced by salt stress were examined by using pharmacological, spectrophotographic and laser scanning confocal microscopic (LSCM) approaches in Vicia faba L. Salt stress resulted in stomatal closure, and this effect was blocked by H2S modulators hypotaurine (HT), aminooxy acetic acid (AOA), hydroxylamine (NH2OH), potassium pyruvate (C3H3KO3) and ammonia (NH3) and H2O2 modulators ascorbic acid (ASA), catalase (CAT), diphenylene iodonium (DPI). Additionally, salt stress induced H2S generation and increased L-/D-cysteine desulfhydrase (L-/D-CDes, pyridoxalphosphate-dependent enzyme) activity in leaves, and caused H2O2 production in guard cells, and these effects were significantly suppressed by H2S modulators and H2O2 modulators respectively. Moreover, H2O2 modulators suppressed salt stress-induced increase of H2S levels and L-/D-CDes activity in leaves as well as stomatal closure of V. faba. However, H2S modulators had no effects on salt stress-induced H2O2 production in guard cells. Altogether, our data suggested that H2S and H2O2 probably are involved in salt stress-induced stomatal closure, and H2S may function downstream of H2O2 in salt stress-induced stomatal movement in V. faba.

scholarly journals Aquaporins facilitate hydrogen peroxide entry into guard cells to mediate ABA- and pathogen-triggered stomatal closure

2017 ◽  
Vol 114 (34) ◽  
pp. 9200-9205 ◽  
Author(s):  
Olivier Rodrigues ◽  
Ganna Reshetnyak ◽  
Alexandre Grondin ◽  
Yusuke Saijo ◽  
Nathalie Leonhardt ◽  
...  
Keyword(s):  
Hydrogen Peroxide ◽  
Water Status ◽  
Stomatal Closure ◽  
Guard Cells ◽  
Molecular Pattern ◽  
Osmotic Water ◽  
Cellular Context ◽  
Guard Cell Protoplasts

Stomatal movements are crucial for the control of plant water status and protection against pathogens. Assays on epidermal peels revealed that, similar to abscisic acid (ABA), pathogen-associated molecular pattern (PAMP) flg22 requires the AtPIP2;1 aquaporin to induce stomatal closure. Flg22 also induced an increase in osmotic water permeability (Pf) of guard cell protoplasts through activation of AtPIP2;1. The use of HyPer, a genetic probe for intracellular hydrogen peroxide (H2O2), revealed that both ABA and flg22 triggered an accumulation of H2O2 in wild-type but not pip2;1 guard cells. Pretreatment of guard cells with flg22 or ABA facilitated the influx of exogenous H2O2. Brassinosteroid insensitive 1-associated receptor kinase 1 (BAK1) and open stomata 1 (OST1)/Snf1-related protein kinase 2.6 (SnRK2.6) were both necessary to flg22-induced Pf and both phosphorylated AtPIP2;1 on Ser121 in vitro. Accumulation of H2O2 and stomatal closure as induced by flg22 was restored in pip2;1 guard cells by a phosphomimetic form (Ser121Asp) but not by a phosphodeficient form (Ser121Ala) of AtPIP2;1. We propose a mechanism whereby phosphorylation of AtPIP2;1 Ser121 by BAK1 and/or OST1 is triggered in response to flg22 to activate its water and H2O2 transport activities. This work establishes a signaling role of plasma membrane aquaporins in guard cells and potentially in other cellular context involving H2O2 signaling.

Inhibition of abscisic acid-induced stomatal closure by ethylene is related to the change of hydrogen peroxide levels in guard cells in broad bean

2011 ◽  
Vol 59 (8) ◽  
pp. 781 ◽  
Author(s):  
XiGui Song ◽  
XiaoPing She ◽  
Juan Wang
Keyword(s):  
Hydrogen Peroxide ◽  
Ascorbic Acid ◽  
Abscisic Acid ◽  
Nadph Oxidase ◽  
Broad Bean ◽  
Stomatal Closure ◽  
Guard Cells ◽  
Diphenylene Iodonium ◽  
Scavenging Enzymes ◽  
Exogenous H2o2

We analysed the role and relationship between hydrogen peroxide (H2O2) reduction and the inhibition of abscisic acid (ABA)-induced stomatal closure by ethylene. Like ascorbic acid (ASA), the most important reducing substrate for H2O2 removal, catalase, one of the H2O2 scavenging enzymes and diphenylene iodonium, an inhibitor of the H2O2-generating enzyme NADPH oxidase, both ethylene-releasing compound 2-chloroethylene phosphonic acid (ethephon, ETH) and 1-aminocyclopropane-1-carboxylic acid (ACC), the immediate precursor of ethylene, were found to inhibit stomatal closure by ABA and to reduce H2O2 levels by ABA in guard cells, indicating that ethylene-caused inhibition of ABA-induced stomatal closure involves reduction of H2O2 levels in guard cells. Additionally, similar to ASA and catalase, ACC/ETH not only suppressed H2O2-induced stomatal closure and H2O2 levels in guard cells treated with exogenous H2O2 in light, but also reopened the stomata which had been closed by ABA and reduced H2O2 levels that had been generated by ABA. The abovementioned effects of ACC and ETH were dissimilar to that of diphenylene iodonium, an inhibitor of the H2O2-generating enzyme NADPH oxidase, which not only had incapability to reduce H2O2 levels by exogenous H2O2 but also could not abolish H2O2 that had been generated by ABA. So we suggest that ethylene probably induces H2O2 removal and reduces H2O2 levels in Vicia faba guard cells, and finally inhibits stomatal closure induced by ABA.

scholarly journals Differences between apoplastic and cytosolic reactive oxygen species production in Arabidopsis during pattern-triggered immunity

2021 ◽  
Author(s):  
Dominique Arnaud ◽  
Michael J. Deeks ◽  
Nicholas Smirnoff
Keyword(s):  
Reactive Oxygen Species ◽  
Hydrogen Peroxide ◽  
Cell Wall ◽  
Stomatal Closure ◽  
Reactive Oxygen ◽  
Guard Cells ◽  
Oxygen Species ◽  
Nadph Oxidases ◽  
Molecular Patterns ◽  
Intracellular Hydrogen Peroxide

AbstractDespite an ever-increasing interest in reactive oxygen species (ROS) signalling during plant-microbe interactions, very little information exists, mainly for technical reasons, on the molecular mechanisms regulating intracellular hydrogen peroxide (H2O2) signalling during PAMP-triggered immunity. Here, we used a sensitive fluorimetry method and the H2O2 sensor roGFP2-Orp1, which revealed unsuspected features on the regulation of cytoplasmic H2O2 and thiol redox dynamics upon pathogen-associated molecular patterns (PAMPs) perception by Arabidopsis thaliana. Extended PAMP-induced cytosolic roGFP2-Orp1 oxidation was distinct from the transient oxidative burst in the apoplast measured by luminol oxidation. Pharmacological and genetic analyses indicate that the prolonged PAMP-induced H2O2 increase in the cytoplasm was largely independent on NADPH oxidases and apoplastic peroxidases. By contrast, the NADPH oxidase mutant rbohF was hyper-sensitive to roGFP2-Orp1 oxidation by H2O2 and PAMP indicating a lower antioxidant capacity. Unlike previous reports, the rbohF mutant, but not rbohD, was impaired in PAMP-triggered stomatal closure and ROS production measured by a fluorescein-based probe in guard cells resulting in defects in stomatal defences against bacteria. However, stomatal closure was not correlated with an increase in roGFP2-Orp1 oxidation in guard cells. Interestingly, RBOHF also participated in PAMP-induced apoplastic alkalinisation. Altogether, our results provide novel insights on the interplay between apoplastic and cytosolic ROS dynamics and highlight the importance of RBOHF in plant immunity.Significance statementPlants mount defence responses to pathogens by detecting pathogen-associated molecular patterns (PAMPs). One response is a rapid and transient burst of reactive oxygen species (ROS, e.g. superoxide and hydrogen peroxide) in the cell wall (apoplast) produced by NADPH oxidases and cell wall peroxidases. Using a genetically-encoded hydrogen peroxide sensor roGFP2-Orp1, we found that, in contrast to the transient apoplastic ROS burst, there is also prolonged hydrogen peroxide production in the cytosol upon PAMP perception which is independent of NADPH oxidase and cell wall peroxidases. Our results suggest that apoplastic ROS rather than intracellular hydrogen peroxide is a signal triggering stomatal closure during PAMP-triggered immunity. Additionally, we re-address the relative contribution of the NADPH oxidases D and F in stomatal immunity.

scholarly journals Ozone Altered Stomatal/Guard Cell Function: Whole Plant and Single Cell Analysis

2000 ◽  
Author(s):  
Eva J. Pell ◽  
Sarah M. Assmann ◽  
Amnon Schwartz ◽  
Hava Steinberger
Keyword(s):  
Hydrogen Peroxide ◽  
Gas Exchange ◽  
Vicia Faba ◽  
Guard Cell ◽  
Cell Function ◽  
Stomatal Closure ◽  
Relative Sensitivity ◽  
Guard Cells ◽  
Stomatal Opening ◽  
K Channels

Original objectives (revisions from original proposal are highlighted) 1. Elucidate the direct effects O3 and H2O2 on guard cell function, utilizing assays of stomatal response in isolated epidermal peels and whole cell gas exchange. 2. Determine the mechanistic basis of O3 and H2O2 effects on the plasma membrane through application of the electrophysiological technique of patch clamping to isolated guard cells. 3. Determine the relative sensitivity of Israeli cultivars of economically important crops to O3 and determine whether differential leaf conductance responses to O3 can explain relative sensitivity to the air pollutant: transfer of technological expertise to Israel. Background to the topic For a long time O3 has been known to reduce gas exchange in plants; it has however been unclear if O3 can affect the stomatal complex directly. Ion channels are essential in stomatal regulation, but O3 has never before been shown to affect these directly. Major conclusions, solution, achievements 1. Ozone inhibits light-induced stomatal opening in epidermal peels isolated from Vicia faba, Arabidopsis thaliana and Nicotiana tabacum in V. faba plants this leads to reduced assimilation without a direct effect on the photosynthetic apparatus. Stomatal opening is more sensitive to O3 than stomatal closure. 2. Ozone causes inhibition of inward K+ channels (involved in stomatal opening) while no detectable effect is observed o the outward K+ channels (stomatal closure). 3. Hydrogen peroxide inhibits stomatal opening and induces stomatal closure in epidermal peels isolated from Vicia faba. 4. Hydrogen peroxide enhances stomatal closure by increasing K+ efflux from guard cells via outward rectifying K+ channels. 5. Based on epidermal peel experiments we have indirectly shown that Ca2+ may play a role in the guard cell response to O3. However, direct measurement of the guard cell [Ca2+]cyt did not show a response to O3. 6. Three Israeli cultivars of zucchini, Clarita, Yarden and Bareqet, were shown to be relatively sensitive to O3 (0.12 ml1-1 ). 7. Two environmentally important Israeli pine species are adversely affected by O3, even at 0.050 ml1-1 , a level frequently exceeded under local tropospheric conditions. P. brutia may be better equipped than P. halepensis to tolerate O3 stress. 8. Ozone directly affects pigment biosynthesis in pine seedlings, as well as the metabolism of O5 precursors, thus affecting the allocation of resources among various metabolic pathways. 9. Ozone induces activity of antioxidant enzymes, and of ascorbate content i the mesophyll and epidermis cells of Commelina communis L. Implications, both scientific and agricultural We have improved the understanding of how O3 and H2O2 do affect guard cell and stomatal function. We have shown that economical important Israeli species like zucchini and pine are relatively sensitive to O3.

scholarly journals Hydrogen peroxide-induced changes in intracellular pH of guard cells precede stomatal closure

Cell Research ◽  
2001 ◽  
Vol 11 (1) ◽  
pp. 37-43 ◽  
Author(s):  
Xiao ZHANG ◽  
Fa Cai DONG ◽  
Jun Feng GAO ◽  
Chun Peng SONG
Keyword(s):  
Hydrogen Peroxide ◽  
Intracellular Ph ◽  
Stomatal Closure ◽  
Guard Cells ◽  
Induced Changes

Hydrogen sulfide induced by hydrogen peroxide mediates brassinosteroid-induced stomatal closure of Arabidopsis thaliana

2021 ◽  
Vol 48 (2) ◽  
pp. 195 ◽  
Author(s):  
Yinli Ma ◽  
Luhan Shao ◽  
Wei Zhang ◽  
Fengxi Zheng
Keyword(s):  
Hydrogen Peroxide ◽  
Arabidopsis Thaliana ◽  
Hydrogen Sulfide ◽  
Stomatal Closure ◽  
Guard Cells ◽  
H2o2 Production ◽  
Wild Type ◽  
Synthesis Inhibitor ◽  
In The Wild ◽  
H2s Production

The role of hydrogen sulfide (H2S) and its relationship with hydrogen peroxide (H2O2) in brassinosteroid-induced stomatal closure in Arabidopsis thaliana (L.) Heynh. were investigated. In the present study, 2,4-epibrassinolide (EBR, a bioactive BR) induced stomatal closure in the wild type, the effects were inhibited by H2S scavenger and synthesis inhibitors, and H2O2 scavengers and synthesis inhibitor. However, EBR failed to close the stomata of mutants Atl-cdes, Atd-cdes, AtrbohF and AtrbohD/F. Additionally, EBR induced increase of L-/D-cysteine desulfhydrase (L-/D-CDes) activity, H2S production, and H2O2 production in the wild type, and the effects were inhibited by H2S scavenger and synthesis inhibitors, and H2O2 scavengers and synthesis inhibitor respectively. Furthermore, EBR increased H2O2 levels in the guard cells of AtrbohD mutant, but couldn’t raise H2O2 levels in the guard cells of AtrbohF and AtrbohD/F mutants. Next, scavengers and synthesis inhibitor of H2O2 could significantly inhibit EBR-induced rise of L-/D-CDes activity and H2S production in the wild type, but H2S scavenger and synthesis inhibitors failed to repress EBR-induced H2O2 production. EBR could increase H2O2 levels in the guard cells of Atl-cdes and Atd-cdes mutants, but EBR failed to induce increase of L-/D-CDes activity and H2S production in AtrbohF and AtrbohD/F mutants. Therefore, we conclude that H2S and H2O2 are involved in the signal transduction pathway of EBR-induced stomatal closure. Altogether, our data suggested that EBR induces AtrbohF-dependent H2O2 production and subsequent AtL-CDes-/AtD-CDes-catalysed H2S production, and finally closes stomata in A. thaliana.

Sign in / Sign up

Export Citation Format

Share Document