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1.
Recent studies have demonstrated that hydrogen sulfide (H2S) produced through the activity of l -cysteine desulfhydrase (DES1) is an important gaseous signaling molecule in plants that could participate in abscisic acid (ABA)-induced stomatal closure. However, the coupling of the DES1/H2S signaling pathways to guard cell movement has not been thoroughly elucidated. The results presented here provide genetic evidence for a physiologically relevant signaling pathway that governs guard cell in situ DES1/H2S function in stomatal closure. We discovered that ABA-activated DES1 produces H2S in guard cells. The impaired guard cell ABA phenotype of the des1 mutant can be fully complemented when DES1/H2S function has been specifically rescued in guard cells and epidermal cells, but not mesophyll cells. This research further characterized DES1/H2S function in the regulation of LONG HYPOCOTYL1 (HY1, a member of the heme oxygenase family) signaling. ABA-induced DES1 expression and H2S production are hyper-activated in the hy1 mutant, both of which can be fully abolished by the addition of H2S scavenger. Impaired guard cell ABA phenotype of des1/hy1 can be restored by H2S donors. Taken together, this research indicated that guard cell in situ DES1 function is involved in ABA-induced stomatal closure, which also acts as a pivotal hub in regulating HY1 signaling.  相似文献   

2.
The role and signaling of sphingosine-1-phosphate (S1P) during darkness-induced stomatal closure were examined in Vicia faba. Darkness substantially raised S1P and hydrogen peroxide (H2O2) levels and closed stomata. These darkness effects were significantly suppressed by DL-threo-dihydrosphingosine (DL-threo-DHS) and N,N-dimethylsphingosine (DMS), two inhibitors of long-chain base kinases. Exogenous S1P led to stomatal closure and H2O2 production, and the effects of S1P were largely prevented by the H2O2 modulators ascorbic acid, catalase, and diphenyleneiodonium. These results indicated that S1P mediated darkness-induced stomatal closure by triggering H2O2 production. In addition, DL-threo-DHS and DMS significantly suppressed both darkness-induced cytosolic alkalization in guard cells and stomatal closure. Exogenous S1P caused cytosolic alkalization and stomatal closure, which could be largely abolished by butyric acid. These results demonstrated that S1P synthesis was necessary for cytosolic alkalization during stomatal closure caused by darkness. Furthermore, together with the data described above, inhibition of darkness-induced H2O2 production by butyric acid revealed that S1P synthesis-induced cytosolic alkalization was a prerequisite for H2O2 production during stomatal closure caused by darkness, a conclusion supported by the facts that the pH increase caused by exogenous S1P had a shorter lag and peaked faster than H2O2 levels and that butyric acid prevented exogenous S1P-induced H2O2 production. Altogether, our data suggested that darkness induced S1P synthesis, causing cytosolic alkalization and subsequent H2O2 production, finally leading to stomatal closure.  相似文献   

3.
Nitric oxide (NO) has recently emerged as a second messenger involved in the complex network of signaling events that regulate stomatal closure. Little is known about the signaling events occurring downstream of NO. Previously, we demonstrated the involvement of phospholipase D (PLD) in NO signaling during stomatal closure. PLDδ, one of the 12 Arabidopsis PLDs, is involved in dehydration stress responses. To investigate the role of PLDδ in NO signaling in guard cells, we analyzed guard cells responses using Arabidopsis wild type and two independent pldδ single mutants. In this work, we show that pldδ mutants failed to close the stomata in response to NO. Treatments with phosphatidic acid, the product of PLD activity, induced stomatal closure in pldδ mutants. Abscisic acid (ABA) signaling in guard cells involved H2O2 and NO production, both required for ABA-induced stomatal closure. pldδ guard cells produced similar NO and H2O2 levels as the wild type in response to ABA. However, ABA- or H2O2-induced stomatal closure was impaired in pldδ plants. These data indicate that PLDδ is downstream of NO and H2O2 in ABA-induced stomatal closure.  相似文献   

4.
Ethylene promotes stomatal closure via inducing hydrogen peroxide (H2O2) generation. H2O2 can be catalytically synthesized by several enzymes in plants. Here, by means of stomatal bioassay, the analysis of enzyme activity and using laser-scanning confocal microscopy based on the H2O2-sensitive probe 2′,7′-dichlorodihydrofluorescein diacetate (H2DCF-DA), the roles of copper amine oxidase (CuAO) in ethylene-induced H2O2 production in guard cells and stomatal closure in Vicia faba L. were investigated. 1-aminocyclopropane-1-carboxylic acid (ACC), an immediate precursor of ethylene synthesis, and ethylene gas significantly activated CuAO in intercellular washing fluid from leaves, the production of H2O2 in guard cells, and stomatal closure. These effects of ACC and ethylene gas were largely prevented by both aminoguanidine and 2-bromoethylamine, which are irreversible inhibitors of CuAO. Among major catalyzed and metabolized products of CuAO, only H2O2 could markedly promote stomatal closure and evidently reversed the effect of CuAO inhibitor on stomatal closure by ACC and ethylene gas. The data described above show that CuAO-mediated H2O2 production is involved in ethylene-induced stomatal closure.  相似文献   

5.
The objective of this study was to clarify the relationships among stomatal, residual, and epidermal conductances in determining the flux of SO2 air pollution to leaves. Variations in leaf SO2 and H2O vapor fluxes were determined using four plant species: Pisum sativum L. (garden pea), Lycopersicon esculentum Mill. flacca (mutant of tomato), Geranium carolinianum L. (wild geranium), and Diplacus aurantiacus (Curtis) Jeps. (a native California shrub). Fluxes were measured using the mass-balance approach during exposure to 4.56 micromoles per cubic meter (0.11 microliters per liter) SO2 for 2 hours in a controlled environmental chamber. Flux through adaxial and abaxial leaf surfaces with closed stomata ranged from 1.9 to 9.4 nanomoles per square meter per second for SO2, and 0.3 to 1.3 millimoles per square meter per second for H2O vapor. Flux of SO2 into leaves through stomata ranged from ~0 to 8.5 (dark) and 3.8 to 16.0 (light) millimoles per square meter per second. Flux of H2O vapor from leaves through stomata ranged from ~0 to 0.6 (dark) to 0.4 to 0.9 (light) millimole per square meter per second. Lycopersicon had internal flux rates for both SO2 and H2O vapor over twice as high as for the other species. Stomatal conductance based on H2O vapor flux averaged from 0.07 to 0.13 mole per square meter per second among the four species. Internal conductance of SO2 as calculated from SO2 flux was from 0.04 mole per square meter per second lower to 0.06 mole per square meter per second higher than stomatal conductance. For Pisum, Geranium, and Diplacus stomatal conductance was the same or slightly higher than internal conductance, indicating that, in general, SO2 flux could be predicted from stomatal conductance for H2O vapor. However, for the Lycopersicon mutant, internal leaf conductance was much higher than stomatal conductance, indicating that factors inside leaves can play a significant role in determining SO2 flux.  相似文献   

6.
7.
Joint Action of O(3) and SO(2) in Modifying Plant Gas Exchange   总被引:5,自引:2,他引:3       下载免费PDF全文
The joint action of O3 and SO2 stress on plants was investigated by determining the quantitative relationship between air pollutant fluxes and effects on stomatal conductance. Gas exchange measurements of O3, SO2, and H2O vapor were made for Pisum sativum L. (garden pea). Plants were grown under controlled environments, and O3, SO2, and H2O vapor fluxes were evaluated with a whole-plant gas exchange chamber using the mass-balance approach. Maximum O3 and SO2 fluxes per unit area (2 sided) into leaves averaged 8 nanomoles per square meter per second with exposure to either O3 or SO2 at 0.1 microliters per liter. Internal fluxes of either O3 or SO2 were reduced by up to 50% during exposure to combined versus individual pollutants; the greatest reduction occurred with simultaneous versus sequential combinations of the pollutants. Stomatal conductance to H2O was substantially altered by the pollutant exposures, with O3 molecules twice as effective as SO2 molecules in inducing stomatal closure. Stomatal conductance was related to the integrated dose of pollutants. The regression equations relating integrated dose to stomatal conductance were similar with O3 alone, O3 plus added SO2, and O3 plus SO2 simultaneously; i.e. a dose of 100 micromoles per square meter produced a 39 to 45% reduction in conductance over nonexposed plants. With SO2 alone, or SO2 plus added O3, a dose of 100 micromoles per square meter produced a 20 to 25% reduction in conductance. When O3 was present at the start of the exposure, then stomatal response resembled that for O3 more than the response for SO2. This study indicated that stomatal responses with combinations of O3 and SO2 are not dependent solely on the integrated dose of pollutants, but suggests that a metabolic synergistic effect exists.  相似文献   

8.
9.
Reactive oxygen species (ROS), including hydrogen peroxide (H2O2), are among the important second messengers in abscisic acid (ABA) signaling in guard cells. In this study, to investigate specific roles of H2O2 in ABA signaling in guard cells, we examined the effects of mutations in the guard cell-expressed catalase (CAT) genes, CAT1 and CAT3, and of the CAT inhibitor 3-aminotriazole (AT) on stomatal movement. The cat3 and cat1 cat3 mutations significantly reduced CAT activities, leading to higher basal level of H2O2 in guard cells, when assessed by 2′,7′-dichlorodihydrofluorescein, whereas they did not affect stomatal aperture size under non-stressed condition. In addition, AT-treatment at concentrations that abolish CAT activities, showed trivial affect on stomatal aperture size, while basal H2O2 level increased extensively. In contrast, cat mutations and AT-treatment potentiated ABA-induced stomatal closure. Inducible ROS production triggered by ABA was observed in these mutants and wild type as well as in AT-treated guard cells. These results suggest that ABA-inducible cytosolic H2O2 elevation functions in ABA-induced stomatal closure, while constitutive increase of H2O2 do not cause stomatal closure.  相似文献   

10.
Specific cellular components have been identified to function in abscisic acid (ABA) regulation of stomatal apertures, including calcium, the cytoskeleton, and phosphatidic acid. In this study, the regulation and dynamic organization of microtubules during ABA-induced stomatal closure by phospholipase D (PLD) and its product PA were investigated. ABA induced microtubule depolymerization and stomatal closure in wide-type (WT) Arabidopsis, whereas these processes were impaired in PLD mutant (pldα1). The microtubule-disrupting drugs oryzalin or propyzamide induced microtubule depolymerization, but did not affect the stomatal aperture, whereas their co-treatment with ABA resulted in stomatal closure in both WT and pldα1. In contrast, the microtubule-stabilizing drug paclitaxel arrested ABA-induced microtubule depolymerization and inhibited ABA-induced stomatal closure in both WT and pldα1. In pldα1, ABA-induced cytoplasmic Ca2+ ([Ca2+]cyt) elevation was partially blocked, and exogenous Ca2+-induced microtubule depolymerization and stomatal closure were impaired. These results suggested that PLDα1 and PA regulate microtubular organization and Ca2+ increases during ABA-induced stomatal closing and that crosstalk among signaling lipid, Ca2+, and microtubules are essential for ABA signaling.  相似文献   

11.
Recent evidence has demonstrated that both copper amine oxidase (CuAO; EC 1.4.3.6) and phospholipase D (PLD; EC 3.1.4.4) are involved in abscisic acid (ABA)-induced stomatal closure. In this study, we investigated the interaction between CuAO and PLD in the ABA response. Pretreatment with either CuAO or PLD inhibitors alone or that with both additively led to impairment of ABA-induced H2O2 production and stomatal closure in Vicia faba. ABA-stimulated PLD activation could not be inhibited by the CuAO inhibitor, and CuAO activity was not affected by the PLD inhibitor. These data suggest that CuAO and PLD act independently in the ABA response. To further examine PLD and CuAO activities in ABA responses, we used the Arabidopsis mutants cuaoζ and pldα1. Ablation of guard cell-expressed CuAOζ or PLDα1 gene retarded ABA-induced H2O2 generation and stomatal closure. As a product of PLD, phosphatidic acid (PA) substantially enhanced H2O2 production and stomatal closure in wide type, pldα1, and cuaoζ. Moreover, putrescine (Put), a substrate of CuAO as well as an activator of PLD, induced H2O2 production and stomatal closure in WT but not in both mutants. These results suggest that CuAO and PLD act independently in ABA-induced stomatal closure.  相似文献   

12.
To ascertain the effect of exogenously applied hydrogen peroxide (H2O2) on drought stress, we examined whether the spraying of soybean leaves with H2O2 would alleviate the symptoms of drought stress. Pre-treatment by spraying leaves with H2O2 delayed foliar wilting caused by drought stress compared to leaves sprayed with distilled water (DW). Additionally, the relative water content of drought-stressed leaves pre-treated with H2O2 was higher than that of leaves pre-treated with DW. Therefore, we analyzed the effect of H2O2 spraying on photosynthetic parameters and on the biosynthesis of oligosaccharides related to water retention in leaves during drought stress. Under conditions of drought stress, the net photosynthetic rate and stomatal conductance of leaves pre-treated with H2O2 were higher than those of leaves pre-treated with DW. In contrast to DW spraying, H2O2 spraying immediately caused an increase in the mRNA levels of d-myo-inositol 3-phosphate synthase 2 (GmMIPS2) and galactinol synthase (GolS), which encode key enzymes for the biosynthesis of oligosaccharides known to help plants tolerate drought stress. In addition, the levels of myo-inositol and galactinol were higher in H2O2-treated leaves than in DW-treated leaves. These results indicated that H2O2 spraying enabled the soybean plant to avoid drought stress through the maintenance of leaf water content, and that this water retention was caused by the promotion of oligosaccharide biosynthesis rather than by rapid stomatal closure.  相似文献   

13.
Effects of hydrogen sulfide (H2S) on plant physiology have been previously studied, but such studies have relied on the use of NaSH as a method for supplying H2S to tissues. Now new compounds which give a less severe H2S shock and a more prolonged exposure to H2S have been developed. Here the effects of one such compound, GYY4137, has been investigated to determine its effects on stomatal closure in Arabidopsis thaliana. It was found that both NaSH and GYY4137 caused stomatal opening in the light and prevented stomatal closure in the dark. Nitric oxide (NO) has been well established as a mediator of stomatal movements and here it was found that both NaSH and GYY4137 reduced the accumulation of NO in guard cells, perhaps suggesting a mode of action for H2S in this system. GYY4137, and future related compounds, will be important tools to unravel the effects of plant exposure to H2S and to determine how H2S may fit into plant cell signalling pathways.  相似文献   

14.
In Cucurbitaceae young leaves are resistant to injury from acute exposure to SO2, whereas mature leaves are sensitive. After exposure of cucumber (Cucumis sativus L.) plants to SO2 at injurious concentrations, illuminated leaves emit volatile sulfur, which is solely H2S. Young leaves emit H2S many times more rapidly than do mature leaves. Young leaves convert approximately 10% of absorbed [35S]SO2 to emitted [35S]H2S, but mature leaves convert less than 2%. These results suggest that a high capability for the reduction of SO2 to H2S and emission of the H2S is a part of the biochemical basis of the resistance of young leaves to SO2.  相似文献   

15.
The effects of chitosan (β-1,4 linked glucosamine, a fungal elicitor), on the patterns of stomatal movement and signaling components were studied. cPTIO (NO scavenger), sodium tungstate (nitrate reductase inhibitor) or l-NAME (NO synthase inhibitor) restricted the chitosan induced stomatal closure, demonstrating that NO is an essential factor. Similarly, catalase (H2O2 scavenger) or DPI [NAD(P)H oxidase inhibitor] and BAPTA-AM or BAPTA (calcium chelators) prevented chitosan induced stomatal closure, suggesting that reactive oxygen species (ROS) and calcium were involved during such response. Monitoring the NO and ROS production in guard cells by fluorescent probes (DAF-2DA and H2DCFDA) indicated that on exposure to chitosan, the levels of NO rose after only 10 min, while those of ROS increased already by 5 min. cPTIO or sodium tungstate or l-NAME prevented the rise in NO levels but did not restrict the ROS production. In contrast, catalase or DPI restricted the chitosan-induced production of both ROS and NO in guard cells. The calcium chelators, BAPTA-AM or BAPTA, did not have a significant effect on the chitosan induced rise in NO or ROS. We propose that the production of NO is an important signaling component and participates downstream of ROS production. The effects of chitosan strike a marked similarity with those of ABA or MJ on guard cells and indicate the convergence of their signal transduction pathways leading to stomatal closure. Nupur Srivastava and Vijay K. Gonugunta have contributed equally.  相似文献   

16.
Nitric oxide, produced from exogenous NO donor, sodium nitroprusside, and hydrogen peroxide exerted antagonistic effects on tobacco leaves at micromolar concentrations but induced synergistic effects at millimolar concentrations. During H2O2-induced oxidative stress, low concentrations of NO inhibited lipid peroxidation, counteracted the fragmentation of total DNA, and prevented accumulation of soluble proteins in Nicotiana plumbaginifolia cells. When applied at high concentrations, NO induced the caspase-like activity, promoted degradation of DNA and soluble proteins, and reduced ATP synthesis. The results are consistent with the hypothesis that NO performs a dual role in plants, acting as antioxidant (scavenger of reactive oxygen species) and as a signaling messenger. There are grounds to believe that, irrespective of the mechanism involved, nitric oxide performs a protective role during oxidative stress in tobacco leaves, because even high concentrations of NO exerted no immediate toxic effect but induced the programmed cell death through the activation of caspase-like proteases.  相似文献   

17.
Low sink demand provided by pod removal and stem girdling of beans (Vicia faba, cv. Daqingshan) (-Sink) induced a significantly lower net photosynthetic rate (P n), stomatal conductance (g s), internal CO2 concentration (C i), and transpiration rate (E) compared with pod and root sink retention (CK). This depression in P n was due to stomatal limitation. Low sink demand of -Sink plants resulted in a higher leaf sucrose content, but a lower sucrose content in guard cells. Moreover, the significant accumulation of H2O2 and ABA were observed in both leaves and guard cells of -Sink plants. The most intensive electron dense deposit of cerium perhydroxides, produced by H2O2 reaction with cerium chloride, was present in the cell walls, especially the dorsal walls of guard cells. Immunogold electron-microscopy localization of ABA showed that ABA was distributed in ventral walls of guard cells and the intercellular space of mesophyll cells of -Sink leaves in contrast to CK plants. Application of exogenous sucrose to isolated bean leaves increased H2O2 and ABA contents. H2O2 and ABA in leaves was likely generated by two independently regulated pathways, each affected by the high sucrose concentration induced by low sink demand. Increased sucrose in leaves in response to low sink demand may have caused the increase of H2O2 and ABA, and their accumulation in mesophyll cells and guard cells was likely the primary cause for stomatal closure under low sink demand.  相似文献   

18.
Fusicoccin (FC) treatment prevents dark‐induced stomatal closure, the mechanism of which is still obscure. By using pharmacological approaches and laser‐scanning confocal microscopy, the relationship between FC inhibition of dark‐induced stomatal closure and the hydrogen peroxide (H2O2) levels in guard cells in broad bean was studied. Like ascorbic acid (ASA), a scavenger of H2O2 and diphenylene iodonium (DPI), an inhibitor of H2O2‐generating enzyme NADPH oxidase, FC was found to inhibit stomatal closure and reduce H2O2 levels in guard cells in darkness, indicating that FC‐caused inhibition of dark‐induced stomatal closure is related to the reduction of H2O2 levels in guard cells. Furthermore, like ASA, FC not only suppressed H2O2‐induced stomatal closure and H2O2 levels in guard cells treated with H2O2 in light, but also reopened the stomata which had been closed by darkness and reduced the level of H2O2 that had been generated by darkness, showing that FC causes H2O2 removal in guard cells. The butyric acid treatment simulated the effects of FC on the stomata treated with H2O2 and had been closed by dark, and on H2O2 levels in guard cells of stomata treated with H2O2 and had been closed by dark, and both FC and butyric acid reduced cytosol pH in guard cells of stomata treated with H2O2 and had been closed by dark, which demonstrates that cytosolic acidification mediates FC‐induced H2O2 removal. Taken together, our results provide evidence that FC causes cytosolic acidification, consequently induces H2O2 removal, and finally prevents dark‐induced stomatal closure.  相似文献   

19.
Shoots of poplar (Populus euramericana L. cv. Flevo) were exposed to filtered air, SO2, NH3 or a mixture of SO2 and NH3 for 7 weeks in fumigation chambers. After this exposure gas exchange measurements were carried out using a leaf chamber. As compared to leaves exposed to filtered air, leaves pretreated with 112 μg m?3 SO2 showed a small reduction in maximum CO2 assimilation rate (Pmax) and stomatal conductance (gs). They also showed a slightly higher quantum yield and dark respiration. In addition, the fluorescence measurements indicated that the Calvin cycle of the leaves pretreated with 112 μg m?3 SO2 was more rapidly activated after transition from dark to light. An exposure to 64 μg m?3 NH3 had a positive effect on Pmax, stomatal conductance and NH3 uptake of the leaves. This positive effect was counteracted by an SO2 concentration of 45 μg m?3. The exposure treatments appeared to have no effect on the relationship between net CO2-assimilation and gs. Also, no injury of the leaf cuticle or of epidermal cells was observed. Resistance analysis showed that NH3 transfer into the leaf can be estimated from data on the boundary layer and stomatal resistance for H2O transfer and NH3 concentration at the leaf surface, irrespective of whether the leaves are exposed for a short or long time to NH3 or to a mixture of NH3 and SO2. In contrast SO2 uptake into the leaves was only partly correlated to the stomatal resistance. The results suggest a large additional uptake of this gas by the leaves. The possibility of a difference in path length between SO2 and H2O molecules is proposed.  相似文献   

20.
UV-B对拟南芥叶片不同来源H2O2的活化和气孔关闭的诱导   总被引:1,自引:0,他引:1  
在UV-B调控植物许多生理过程中过氧化氢(H2O2)作为第二信使发挥着重要作用,但H2O2来源途径并不清楚。该研究借助气孔开度分析和激光扫描共聚焦显微镜技术,探讨H2O2在介导不同剂量UV-B诱导拟南芥叶片气孔关闭过程中的酶学来源途径。结果发现:0.5W.m-2 UV-B能诱导野生型拟南芥叶片保卫细胞的H2O2产生和气孔关闭,且该效应能被NADPH氧化酶抑制剂二苯基碘(DPI)抑制,而不能被细胞壁过氧化物酶抑制剂水杨基氧肟酸(SHAM)抑制,同时该剂量UV-B也不能诱导NADPH氧化酶功能缺失单突变体AtrbohD和AtrbohF以及双突变体AtrbohD/F保卫细胞的H2O2产生和气孔关闭;相反,0.65 W.m-2 UV-B既能诱导野生型也能诱导NADPH氧化酶突变体保卫细胞的H2O2产生和气孔关闭,且该效应能被SHAM抑制,却不能被DPI抑制。结果表明,不同剂量UV-B通过活化不同生成途径的H2O2来诱导拟南芥叶片气孔关闭,即低剂量UV-B主要诱导NADPH氧化酶AtrbohD和AtrbohF途径来源的H2O2生成,而高剂量UV-B主要活化细胞壁过氧化酶途径来源的H2O2。  相似文献   

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