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

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.

2019 ◽  
Author(s):  
Yinli Ma ◽  
Luhan Shao ◽  
Jiao Niu

Abstract Background Whether hydrogen sulfide (H2S) mediates darkness-induced stomatal closure in A. thaliana is unknown, and the interaction between H2S and hydrogen peroxide (H2O2) in the process needs to be elucidated. Results Our results indicated that H2S modulators hypotaurine (HT), aminooxy acetic acid (AOA), hydroxylamine (NH2OH) and potassium pyruvate (N3H3KO3)+ammonia (NH3) all inhibited darkness-induced stomatal closure, H2S generation and L-/D-cysteine desulfhydras (L-/D-CDes) activity increase in wild-type A. thaliana leaves. Darkness induced stomatal closure in wild-type plants, but failed in Atl-cdes and Atd-cdes mutants. Additionally, H2S content and L-/D-CDes activity were significantly decreased after application with H2O2 modulators ascorbic acid (ASA), catalase (CAT), diphenylene iodonium (DPI), and salicylhydroxamic acid (SHAM) in darkness, but there is almost no effects on H2O2 levels when in presence of HT, AOA, NH2OH, and C3H3KO3+NH3 in darkness in wild-type plants. Moreover, darkness couldn't increase H2S content and L-/D-CDes activity of AtrbohF and AtrbohD/F mutants leaves, but the levels of H2O2 increased in guard cells of Atl-cdes and Atd-cdes mutants. Conclusions The results suggest that L-/D-CDes-generated H2S mediates darkness-induced stomatal closure, and functions downstream of H2O2 in A. thaliana.


2019 ◽  
Author(s):  
Yinli Ma ◽  
Luhan Shao ◽  
Jiao Niu

Abstract Background Whether stomatal movement by darkness in Arabidopsis thaliana is mediated by hydrogen sulfide (H2S) is undiscovered yet, so the interaction between hydrogen peroxide (H2O2) and H2S in the process needs to be elucidated. Results Our results indicated that H2S modulators aminooxy acetic acid (AOA), potassium pyruvate (N3H3KO3) + ammonia (NH3), hydroxylamine (NH2OH), and hypotaurine (HT) inhibited darkness-induced stomatal closure, H2S generation and L-/D-cysteine desulfhydrase (L-/D-CDes) activity increased in wild-type A. thaliana leaves. Darkness induced stomatal closure in wild-type plants, but failed in Atl-cdes and Atd-cdes mutants. Additionally, both L-/D-CDes activity and H2S content were significantly decreased after applying H2O2 modulators salicylhydroxamic acid (SHAM), ascorbic acid (ASA), diphenylene iodonium (DPI), and catalase (CAT) in darkness, but there was almost no effects on H2O2 levels in the presence of AOA, C3H3KO3+NH3, NH2OH, and HT of wild-type plants in darkness. Moreover, darkness couldn't increase H2S content and L-/D-CDes activity of AtrbohF and AtrbohD/F mutants leaves, but increased H2O2 levels in Atl-cdes and Atd-cdes guard cells. Conclusions We observed that L-/D-CDes-generated H2S mediates stomatal closure by darkness, and functions downstream of H2O2 in A. thaliana.


2019 ◽  
Vol 46 (2) ◽  
pp. 136 ◽  
Author(s):  
Yinli Ma ◽  
Wei Zhang ◽  
Jiao Niu ◽  
Yu Ren ◽  
Fan Zhang

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.


Author(s):  
Md Tahjib-Ul-Arif ◽  
Shintaro Munemasa ◽  
Toshiyuki Nakamura ◽  
Yoshimasa Nakamura ◽  
Yoshiyuki Murata

Abstract Cytosolic calcium ([Ca2+]cyt) elevation activates plasma membrane anion channels in guard cells, which is required for stomatal closure. However, involvement of the anion channels in the [Ca2+]cyt elevation remains unclear. We investigated the involvement using Arabidopsis thaliana anion channel mutants, slac1-4 slah3-3 and slac1-4 almt12-1. Extracellular calcium induced stomatal closure in the wild-type plants but not in the anion channel mutant plants whereas extracellular calcium induced [Ca2+]cyt elevation both in the wild-type guard cells and in the mutant guard cells. The peak height and the number of the [Ca2+]cyt spike were lower and larger in the slac1-4 slah3-3 than in the wild-type and the height and the number in the slac1-4 almt12-1 were much lower and much larger than in the wild-type. These results suggest that the anion channels are involved in the regulation of [Ca2+]cyt elevation in guard cells.


2014 ◽  
Vol 41 (8) ◽  
pp. 803 ◽  
Author(s):  
Xiao-Min Ge ◽  
Yan Zhu ◽  
Jun-Min He

The role and the interrelationship of cytosolic alkalisation and nitric oxide (NO) in UVB-induced stomatal closure were investigated in Arabidopsis thaliana (L.) Heynh. by stomatal bioassay and laser-scanning confocal microscopy. In response to 0.5 W m–2 UVB radiation, the rise of NO levels in guard cells occurred after cytosolic alkalisation but preceded stomatal closure. UVB-induced NO production and stomatal closure were both inhibited by NO scavengers, nitrate reductase (NR) inhibitors and a Nia2–5/Nia1–2 mutation, and also by butyrate. Methylamine induced NO generation and stomatal closure in the wild-type but not in the Nia2–5/Nia1–2 mutant or wild-type plants pretreated with NO scavengers or NR inhibitors while enhancing the cytosolic pH in guard cells under light. NO generation in wild-type guard cells was largely induced after 60 min of UVB radiation. The defect in UVB-induced NO generation in Nia2–5/Nia1–2 guard cells did not affect the changes of guard cell pH before 60 min of UVB radiation, but prevented the UVB-induced cytosolic alkalisation after 60 min of radiation. Meanwhile, exogenous NO caused a marked rise of cytosolic pH in guard cells. Together, our results show that cytosolic alkalisation and NR-dependent NO production coordinately function in UVB signalling in A. thaliana guard cells.


2020 ◽  
Vol 61 (10) ◽  
pp. 1788-1797
Author(s):  
Md. Moshiul Islam ◽  
Wenxiu Ye ◽  
Fahmida Akter ◽  
Mohammad Saidur Rhaman ◽  
Daiki Matsushima ◽  
...  

Abstract Production of reactive oxygen species (ROS) is a key signal event for methyl jasmonate (MeJA)- and abscisic acid (ABA)-induced stomatal closure. We recently showed that reactive carbonyl species (RCS) stimulates stomatal closure as an intermediate downstream of hydrogen peroxide (H2O2) production in the ABA signaling pathway in guard cells of Nicotiana tabacum and Arabidopsis thaliana. In this study, we examined whether RCS functions as an intermediate downstream of H2O2 production in MeJA signaling in guard cells using transgenic tobacco plants overexpressing A. thaliana 2-alkenal reductase (n-alkanal + NAD(P)+ ⇌ 2-alkenal + NAD(P)H + H+) (AER-OE tobacco) and Arabidopsis plants. The stomatal closure induced by MeJA was impaired in the AER-OE tobacco and was inhibited by RCS scavengers, carnosine and pyridoxamine, in the wild-type (WT) tobacco plants and Arabidopsis plants. Application of MeJA significantly induced the accumulation of RCS, including acrolein and 4-hydroxy-(E)-2-nonenal, in the WT tobacco but not in the AER-OE plants. Application of MeJA induced H2O2 production in the WT tobacco and the AER-OE plants and the H2O2 production was not inhibited by the RCS scavengers. These results suggest that RCS functions as an intermediate downstream of ROS production in MeJA signaling and in ABA signaling in guard cells.


Cells ◽  
2021 ◽  
Vol 10 (12) ◽  
pp. 3399
Author(s):  
Ilaria Fraudentali ◽  
Chiara Pedalino ◽  
Paraskevi Tavladoraki ◽  
Riccardo Angelini ◽  
Alessandra Cona

Plant defence responses to adverse environmental conditions include different stress signalling, allowing plant acclimation and survival. Among these responses one of the most common, immediate, and effective is the modulation of the stomatal aperture, which integrates different transduction pathways involving hydrogen peroxide (H2O2), calcium (Ca2+), nitric oxide (NO), phytohormones and other signalling components. The Arabidopsis thaliana copper amine oxidases β (AtCuAOβ) encodes an apoplastic CuAO expressed in guard cells and root protoxylem tissues which oxidizes polyamines to aminoaldehydes with the production of H2O2 and ammonia. Here, its role in stomatal closure, signalled by the wound-associated phytohormone methyl-jasmonate (MeJA) was explored by pharmacological and genetic approaches. Obtained data show that AtCuAOβ tissue-specific expression is induced by MeJA, especially in stomata guard cells. Interestingly, two Atcuaoβ T-DNA insertional mutants are unresponsive to this hormone, showing a compromised MeJA-mediated stomatal closure compared to the wild-type (WT) plants. Coherently, Atcuaoβ mutants also show compromised H2O2-production in guard cells upon MeJA treatment. Furthermore, the H2O2 scavenger N,N1-dimethylthiourea (DMTU) and the CuAO-specific inhibitor 2-bromoethylamine (2-BrEtA) both reversed the MeJA-induced stomatal closure and the H2O2 production in WT plants. Our data suggest that AtCuAOβ is involved in the H2O2 production implicated in MeJA-induced stomatal closure.


2015 ◽  
Vol 42 (2) ◽  
pp. 126 ◽  
Author(s):  
Suli Yan ◽  
Shuitian Luo ◽  
Shanshan Dong ◽  
Ting Zhang ◽  
Jingru Sun ◽  
...  

Heterotrimeric G-proteins play an important role in plant signalling pathways. The plant hormone methyl jasmonate (MeJA) can induce stomatal closure in many plant species. The signal cascade in MeJA-induced stomatal closure has been studied previously. However, the function of G proteins in this process has not yet been evaluated. In this study, the stomatal movement induced by MeJA in the wild-type Arabidopsis thaliana (L. Heynh.) (WS), Gα subunit loss-of-function mutant gpa1–1 and gpa1–2 guard cells were measured. Further, the transmembrane ion flux (H+, Ca2+ and K+) and reactive oxygen species (ROS) experiments were performed in guard cells from WS, GDP-β-S pre-treated WS, gpa1–1 and gpa1–2 using non-invasive micro-test technique (NMT) and confocal technique. It was observed that the MeJA-induced stomatal closure was abolished in guard cells of gpa1 mutants. GDP-β-S pre-treatment and gpa1 mutants impaired the MeJA-activated H+ efflux, Ca2+ influx and K+ efflux. The accumulation of ROS in gpa1–1 and gpa1–2 guard cells was also lower than that in WS guard cells under MeJA treatment. These results suggested that Gα subunits are involved in regulating the signal events in JA signal pathway and stomatal closure.


2020 ◽  
Vol 11 ◽  
Author(s):  
Xuan-shan Liu ◽  
Chao-chao Liang ◽  
Shu-guo Hou ◽  
Xin Wang ◽  
Dong-hua Chen ◽  
...  

Guard cells shrink in response to drought stress and abscisic acid (ABA) signaling, thereby reducing stomatal aperture. Hydrogen peroxide (H2O2) is an important signaling molecule acting to induce stomatal closure. As yet, the molecular basis of control over the level of H2O2 in the guard cells remains largely unknown. Here, the leucine-rich repeat (LRR)—receptor-like kinase (RLK) protein HSL3 has been shown to have the ability to negatively regulate stomatal closure by modulating the level of H2O2 in the guard cells. HSL3 was markedly up-regulated by treating plants with either ABA or H2O2, as well as by dehydration. In the loss-of-function hsl3 mutant, both stomatal closure and the activation of anion currents proved to be hypersensitive to ABA treatment, and the mutant was more tolerant than the wild type to moisture deficit; the overexpression of HSL3 had the opposite effect. In the hsl3 mutant, the transcription of NADPH oxidase gene RbohF involved in H2O2 production showed marked up-regulation, as well as the level of catalase activity was weakly inducible by ABA, allowing H2O2 to accumulate in the guard cells. HSL3 was concluded to participate in the regulation of the response to moisture deficit through ABA-induced stomatal closure triggered by the accumulation of H2O2 in the guard cells.


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