Role and interrelationship of PTPases and H2O2 in light/dark-regulated stomatal movement in Vicia faba

2009 ◽  
Vol 57 (6) ◽  
pp. 486 ◽  
Author(s):  
Yuanhua Zhang ◽  
Xiaoping She ◽  
Guangbin Zhang
Keyword(s):  
Vicia Faba ◽  
Laser Scanning ◽  
Stomatal Closure ◽  
Specific Inhibitor ◽  
Guard Cells ◽  
Stomatal Aperture ◽  
Stomatal Movement ◽  
Obvious Effect ◽  
Exogenous H2o2 ◽  
Ptpase Activity

Role and interrelationship of protein tyrosine phosphatases (PTPases) and H2O2 in light/dark-regulated stomatal movement in Vicia faba were investigated by epidermal strip bioassay, laser-scanning confocal microscopy and assays of PTPase activity. Our results indicate that phenylarsine oxide (PAO), a specific inhibitor of PTPases, ascorbic acid (ASA), an important reducing substrate for H2O2 removal, and catalase (CAT), one of the H2O2 scavenging enzymes, did not cause any change of stomatal aperture in light, but remarkably prevented dark-induced stomatal closure. Exogenous H2O2 had no obvious effect on stomatal aperture in the dark, but significantly induced stomatal closure in light. Both PTPase activity in epidermal strips and endogenous H2O2 level in guard cells in the dark were higher than those in light. The results showed that both PTPases and H2O2 mediate light/dark-regulated stomatal movement, that dark-induced stomatal closure requires the activation of PTPases and the enhancement of H2O2 levels in guard cells, and stomatal opening caused by light is associated with the inactivation of PTPases and the reduction of H2O2 levels in guard cells. Additionally, like ASA and CAT, PAO abolished dark-, exogenous H2O2-induced stomatal closure and dichlorofluorescein fluorescence in guard cells, indicating that activation of PTPases can enhance H2O2 levels probably via suppressing the decrease of H2O2 levels in guard cells. On the other hand, similar to PAO, ASA and CAT evidently prevented dark-, exogenous H2O2-induced stomatal closure and obviously inactivated PTPases in the dark. However, exogenous H2O2 significantly activated PTPases in light. The results show that H2O2 can induce activation of PTPases. Taken together, the present results provide evidence that both H2O2 and PTPases are involved in light/dark-regulated stomatal movement, and the interaction between H2O2 and PTPases plays a pivotal role in light/dark signal transduction process in guard cells.

Cytokinin- and auxin-induced stomatal opening involves a decrease in levels of hydrogen peroxide in guard cells of Vicia faba

2006 ◽  
Vol 33 (6) ◽  
pp. 573 ◽  
Author(s):  
Xi-Gui Song ◽  
Xiao-Ping She ◽  
Jun-Min He ◽  
Chen Huang ◽  
Tu-sheng Song
Keyword(s):  
Hydrogen Peroxide ◽  
Vicia Faba ◽  
Laser Scanning ◽  
Guard Cells ◽  
Laser Scanning Confocal Microscopy ◽  
Stomatal Opening ◽  
Scanning Confocal Microscopy ◽  
Diphenylene Iodonium ◽  
The Relationship ◽  
Exogenous H2o2

Previous studies have shown that cytokinins and auxins can induce the opening of stomata. However, the mechanism of stomatal opening caused by cytokinins and auxins remains unclear. The purpose of this paper is to investigate the relationship between hydrogen peroxide (H2O2) levels in guard cells and stomatal opening induced by cytokinins and auxins in Vicia faba. By means of stomatal bioassay and laser-scanning confocal microscopy, we provide evidence that cytokinins and auxins reduced the levels of H2O2 in guard cells and induced stomatal opening in darkness. Additionally, cytokinins not only reduced exogenous H2O2 levels in guard cells caused by exposure to light, but also abolished H2O2 that had been generated during a dark period, and promoted stomatal opening, as did ascorbic acid (ASA, an important reducing substrate for H2O2 removal). However, unlike cytokinins, auxins did not reduce exogenous H2O2, did not abolish H2O2 that had been generated in the dark, and therefore did not promote reopening of stoma induced to close in the dark. The above-mentioned effects of auxins were similar to that of diphenylene iodonium (DPI, an inhibitor of the H2O2-generating enzyme NADPH oxidase). Taken together our results indicate that cytokinins probably reduce the levels of H2O2 in guard cells by scavenging, whereas auxins limit H2O2 levels through restraining H2O2 generation, inducing stomatal opening in darkness.

The role and the interrelationship of hydrogen peroxide and nitric oxide in the UV-B-induced stomatal closure in broad bean

2005 ◽  
Vol 32 (3) ◽  
pp. 237 ◽  
Author(s):  
Jun-Min He ◽  
Hua Xu ◽  
Xiao-Ping She ◽  
Xi-Gui Song ◽  
Wen-Ming Zhao
Keyword(s):  
Vicia Faba ◽  
Laser Scanning ◽  
Broad Bean ◽  
Stomatal Closure ◽  
Guard Cells ◽  
Laser Scanning Confocal Microscopy ◽  
H2o2 Production ◽  
No Production ◽  
No Donor ◽  
Scanning Confocal Microscopy

Previous studies have showed that UV-B can stimulate closure as well as opening of stomata. However, the mechanism of this complex effect of UV-B is not clear. The purpose of this paper is to investigate the role and the interrelationship of H2O2 and NO in UV-B-induced stomatal closure in broad bean (Vicia faba L.). By epidermal strip bioassay and laser-scanning confocal microscopy, we observed that UV-B-induced stomatal closure could be largely prevented not only by NO scavenger c-PTIO or NO synthase (NOS) inhibitor l-NAME, but also by ascorbic acid (ASC, an important reducing substrate for H2O2 removal) or catalase (CAT, the H2O2 scavenger), and that UV-B-induced NO and H2O2 production in guard cells preceded UV-B-induced stomatal closure. These results indicate that UV-B radiation induces stomatal closure by promoting NO and H2O2 production. In addition, c-PTIO, l-NAME, ASC and CAT treatments could effectively inhibit not only UV-B-induced NO production, but also UV-B-induced H2O2 production. Exogenous H2O2-induced NO production and stomatal closure were partly abolished by c-PTIO and l-NAME. Similarly, exogenous NO donor sodium nitroprusside-induced H2O2 production and stomatal closure were also partly reversed by ASC and CAT. These results show a causal and interdependent relationship between NO and H2O2 during UV-B-regulated stomatal movement. Furthermore, the l-NAME data also indicate that the NO in guard cells of Vicia faba is probably produced by a NOS-like enzyme.

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.

UV-B-induced stomatal closure occurs via ethylene-dependent NO generation in Vicia faba

2011 ◽  
Vol 38 (4) ◽  
pp. 293 ◽  
Author(s):  
Jun-Min He ◽  
Zhan Zhang ◽  
Rui-Bin Wang ◽  
Yi-Ping Chen
Keyword(s):  
Vicia Faba ◽  
Laser Scanning ◽  
Stomatal Closure ◽  
Guard Cells ◽  
Laser Scanning Confocal Microscopy ◽  
Ultraviolet B ◽  
Ethylene Synthesis ◽  
Scanning Confocal Microscopy ◽  
Bean Plants ◽  
Exogenous Ethylene

The role of ethylene and the relationship between ethylene and nitric oxide (NO) in ultraviolet B (UV-B)-induced stomatal closure were investigated in Vicia faba L. (broad bean) plants by epidermal strip bioassay, laser-scanning confocal microscopy and assay of ethylene production. In response to UV-B radiation, the rise of NO level in guard cells was after ethylene evolution peak, but preceded stomatal closure. Both UV-B-induced NO generation in guard cells and subsequent stomatal closure were substantially inhibited not only by NO scavenger and nitrate reductase (NR) inhibitors, but also by interfering with ethylene synthesis or perception. Although exogenous NO could reverse the inhibitive effect of interfering with ethylene synthesis or perception on UV-B-induced stomatal closure, the inhibitive effect of NO scavenger and NR inhibitors on UV-B-induced stomatal closure could not be rescued by exogenous ethylene. Taken together, our results clearly show that ethylene participates in the UV-B-induced stomatal closure and acts upstream of the NR source of NO generation in V. faba.

Pharmacological evidence indicates that MAPKK/CDPK modulate NO levels in darkness-induced stomatal closure of broad bean

2008 ◽  
Vol 56 (4) ◽  
pp. 347 ◽  
Author(s):  
Xiaoping She ◽  
Xigui Song
Keyword(s):  
Nitric Oxide ◽  
Protein Kinase ◽  
Laser Scanning ◽  
Broad Bean ◽  
Stomatal Closure ◽  
Specific Inhibitor ◽  
Guard Cells ◽  
Laser Scanning Confocal Microscopy ◽  
Mitogen Activated Protein Kinase ◽  
Nitric Oxide Synthase Inhibitor

By using pharmacological approaches and laser scanning confocal microscopy (LSCM) based on 4, 5-diaminofluorescein diacetate (DAF-2 DA), the roles of MAPKK/CDPK and their effects on nitric oxide (NO) levels of guard cells during darkness-induced stomatal closure in broad bean were investigated. The results indicated that both 2′-amino-3′-methoxyflavone (PD98059) (an inhibitor of mitogen-activated protein kinase kinase, MAPKK) and trifluoperazine (TFP) (a specific inhibitor of calcium-dependent protein kinase, CDPK) reduced the levels of NO in guard cells and significantly reversed darkness-induced stomatal closure, implying that MAPKK/CDPK mediate darkness-induced stomatal closure by enhancing NO levels in guard cells. In addition, as with NO scavenger 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (cPTIO), but not with nitric oxide synthase inhibitor NG-nitro-L-Arg-methyl ester (L-NAME), PD98059 and TFP not only reduced 4,5-diaminofluorescein diacetate (DAF-2 DA) fluorescence in guard cells by sodium nitroprusside (SNP) in light, but also abolished NO that had been generated during a dark period, and reversed stomatal closure by SNP and by darkness, suggesting MAPKK and CDPK are probably related to restraining the NO scavenging to elevate NO levels in guard cells, during darkness-induced stomatal closure. The results also showed that both PD98059 and TFP reduced stomatal closure by SNP, implying that the possibility of MAPKK and CDPK acting as the target downstream of NO should not be ruled out. There may be a causal and interdependent relationship between MAPKK/CDPK and NO in darkness-induced stomatal closure, and in the process this cross-talk may lead to the formation of a self-amplification loop about them.

Crosstalk of NO with Ca2+ in Stomatal Movement in Vicia faba Guard Cells

2009 ◽  
Vol 35 (8) ◽  
pp. 1491-1499 ◽  
Author(s):  
Lin ZHANG ◽  
Xiang ZHAO ◽  
Ya-Jing WANG ◽  
Xiao ZHANG
Keyword(s):  
Vicia Faba ◽  
Guard Cells ◽  
Stomatal Movement

Actin microfilaments and vacuoles are downstream targets of H2O2 signalling pathways in hyperosmotic stress-induced stomatal closure

2011 ◽  
Vol 38 (4) ◽  
pp. 303
Author(s):  
Ai-Xia Huang ◽  
Xiao-Ping She
Keyword(s):  
Osmotic Pressure ◽  
Laser Scanning ◽  
Stomatal Closure ◽  
Guard Cells ◽  
Laser Scanning Confocal Microscopy ◽  
Oxidase Inhibitor ◽  
Hyperosmotic Stress ◽  
H2o2 Production ◽  
H2o2 Treatment ◽  
Scanning Confocal Microscopy

Changes in osmotic pressure can induce stomatal closure to reduce transpirational water loss from plants. In the present work, we investigated the mechanism underlying the perception and transduction of extracellular changes in osmotic pressure in Vicia faba L. guard cells. Using an epidermal strip bioassay and laser-scanning confocal microscopy, we provide evidence that hyperosmotic stress treatment led to stomatal closure and the rapid promotion of hydrogen peroxide (H2O2) production in V. faba guard cells. The effects were largely reduced by H2O2 scavengers ASA, CAT, NADPH oxidase inhibitor DPI and cell wall peroxidase inhibitor SHAM. These results indicate that hyperosmotic stress induces stomatal closure by promoting H2O2 production. Cytochalasin B (CB), latrunculin B (Lat B) and jasplakinolide (JK) inhibited stomatal closure induced by hyperosmotic stress but didn’t prevent the increase of endogenous H2O2 levels, suggesting that microfilaments reorganisation participates in stomatal closure induced by hyperosmotic stress, and may act downstream of H2O2 signalling processes. In addition, we observed splitting of big vacuoles into many small vacuoles in response to hyperosmotic stress and H2O2 treatment, and CB inhibited these changes of vacuoles; stomatal closure was also inhibited. Taken together these results indicate that the stomatal closure in response to hyperosmotic stress may initiate H2O2 generation, and that reorganisation of microfilaments and the changing of vacuoles occurs downstream of H2O2 signalling processes.

scholarly journals Downregulating VAC14 in Guard Cells Causes Drought Hypersensitivity by Inhibiting Stomatal Closure

2020 ◽  
Vol 11 ◽  
Author(s):  
Zong-Qi Wang ◽  
Qi Liu ◽  
Ju-Hua Wu ◽  
Juan Li ◽  
Jun-Min He ◽  
...  
Keyword(s):  
Stomatal Closure ◽  
Guard Cells ◽  
Land Plant ◽  
Scaffolding Protein ◽  
Stomatal Movement ◽  
Wild Type ◽  
Physiological Factors ◽  
Surrounding Environment ◽  
Intracellular Activities

Stomata are a key land plant innovation that permit the regulation of gaseous exchanges between the plant interior and the surrounding environment. By opening or closing, stomata regulate transpiration of water though the plant; and these actions are coordinated with acquisition of CO2 for photosynthesis. Stomatal movement is controlled by various environmental and physiological factors and associates with multiple intracellular activities, among which the dynamic remodeling of vacuoles plays a crucial role. Phosphatidylinositol 3,5-bisphosphate [PI(3,5)P2] is critical for dynamic remodeling of vacuoles. Its production requires a PI(3,5)P2-metabolizing complex consisting of FAB1/PIKfyve kinases, SAC phosphatases, and the scaffolding protein VAC14. Although genetic or pharmacological downregulation of PI(3,5)P2 causes hyposensitivity to ABA-induced stomatal closure, whether the effect of PI(3,5)P2 on stomatal movement is cell-autonomous and the physiological consequences of its reduction were unclear. We report that downregulating Arabidopsis VAC14 specifically in guard cells by artificial microRNAs (amiR-VAC14) results in enlarged guard cells and hyposensitivity to ABA- and dark-induced stomatal closure. Vacuolar fission during stomatal closure is compromised by downregulating VAC14 in guard cells. Exogenous application of PI(3,5)P2 rescued the amiR-VAC14 phenotype whereas PI(3,5)P2 inhibitor YM201636 caused wild-type plants to have inhibited stomatal closure. We further show that downregulating VAC14 specifically in guard cells impairs drought tolerance, suggestive of a key role of guard cell-produced PI(3,5)P2 in plant fitness.

Cytosolic alkalization-mediated H2O2 and NO production are involved in darkness-induced stomatal closure in Vicia faba

2013 ◽  
Vol 93 (1) ◽  
pp. 119-130 ◽  
Author(s):  
Yinli Ma ◽  
Xiaoping She ◽  
Shushen Yang
Keyword(s):  
Nitric Oxide ◽  
Vicia Faba ◽  
Butyric Acid ◽  
Stomatal Closure ◽  
Guard Cells ◽  
Weak Acid ◽  
No Production ◽  
Tetraacetic Acid ◽  
Acetoxymethyl Ester ◽  
Cytosolic Ph

Ma, Y., She, X. and Yang, S. 2013. Cytosolic alkalization-mediated H 2 O 2 and NO production are involved in darkness-induced stomatal closure in Vicia faba. Can. J. Plant Sci. 93: 119–130. Darkness raised cytosolic pH, hydrogen peroxide (H2O2) and nitric oxide (NO) levels in guard cells while inducing Vicia faba stomatal closure. These darkness effects were prevented by weak acid butyric acid, H2O2 modulators ascorbic acid (ASA), catalase (CAT), diphenyleneiodonium (DPI) and NO modulators 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (c-PTIO), NG-nitro-L-arg-methyl ester (L-NAME) respectively. The data suggest that cytosolic alkalization, H2O2 and NO all participate in darkness-induced stomatal closure. During darkness treatment, pH rise became noticeable at 10 min and peaked at 25 min, while H2O2 and NO production increased significantly at 20 min and reached their maximums at 40 min. The H2O2 and NO levels were increased by methylamine in light and decreased by butyric acid in darkness. The results show that cytosolic alkalization induces H2O2 and NO production. ASA, CAT and DPI suppressed NO production by methylamine, c-PTIO and L-NAME prevented H2O2 generation by methylamine. Calcium chelator 1,2-bis (2-aminophenoxy)ethane-N,N,N′,N′-tetraacetic acid acetoxymethyl ester (BAPTA-AM) and 1,2-bis(2-aminophenoxy)ethane-N,N,N′,N′-tetraacetic acid (BAPTA) restricted darkness-induced alkalization, H2O2 and NO production and stomatal closure. We suggest that cytosolic alkalization is necessary for H2O2 and NO production during darkness-induced stomatal closure. H2O2 mediates NO synthesis by alkalization, and vice versa. Calcium may act upstream of cytosolic alkalization, H2O2 and NO production, besides its known action downstream of H2O2 and NO.

Preparation of rhodamine-labelled tubulin and its reassembly in guard cells in leaves of Vicia faba L.

2004 ◽  
Vol 1 (3) ◽  
pp. 161-165
Author(s):  
Yu Rong ◽  
Yuan Ming ◽  
Wang Xue-Chen
Keyword(s):  
Vicia Faba ◽  
Laser Scanning ◽  
Guard Cells ◽  
Confocal Laser Scanning Microscope ◽  
Confocal Laser ◽  
Dorsal Wall ◽  
Pig Brain ◽  
Brain Tubulin ◽  
Ventral Wall

AbstractTubulin from fresh pig brain was extracted through two cycles of polymerization and depolymerization, purified by chromatography on an ion-exchange column and labelled with 5(6)-carboxytetramethylrhodamine succinimidyl ester. We prepared 10.8 mg/ml rhodamine-labelled tubulin probe (dye/protein=0.6) and then microinjected it into living guard cells in leaves of Vicia faba L. After 5–10 min, a network of microtubules (MTs) could clearly be observed in the guard cells under a confocal laser scanning microscope. In opening guard cells, the cortical MTs radiated from the ventral wall to the dorsal wall and in closed guard cells the radial MTs were completely depolymerized into random and diffuse fragments. This shows that pig brain tubulin can easily be incorporated into MTs in guard cells, and that the dynamic behaviour of MTs can be examined directly in vivo during stomatal movement.

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