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. 2026 May 15;94:104205. doi: 10.1016/j.redox.2026.104205

Aboveground whole-plant live H2S imaging method sheds new light on the relationships between H2S, NO, and H2O2

Devasantosh Mohanty a, María Ángeles Peláez-Vico a, Ranjita Sinha a, María Inmaculada Sánchez-Vicente b, Oscar Lorenzo b, Ron Mittler a,c,⁎
PMCID: PMC13199770  PMID: 42140139

Abstract

Hydrogen peroxide (H2O2), nitric oxide (NO), and hydrogen sulfide (H2S) regulate processes such as growth, development, and stress responses via post-translational modifications that alter the structure, localization, and function of multiple cellular proteins. H2O2, NO, and H2S are also thought to regulate the levels of each other in cells via multiple pathways. Although several methods were developed for their imaging at the cell and tissue levels, whole-plant live imaging methods were only developed for NO and H2O2. Here, we report on the development of a method for whole-plant live imaging of H2S that complements the H2O2 and NO methods we previously developed and can be used side-by-side with them. Using H2S donor and scavenger, the l-CYSTEINE DESULFHYDRASE 1 (des1) mutant, as well as treatment of plants with heat stress (HS), or flg22, we confirmed the specificity and biological relevance of the method developed. Using side-by-side imaging of H2O2, NO and H2S, in wild type and different mutants, following HS, we further reveal that H2O2 accumulation could be uncoupled from H2S and NO accumulation, but that H2S and NO accumulation are mostly tied to each other, suggesting a close interplay between them. We also show that H2S accumulation following HS requires NO accumulation, and that mutants deficient in ASCORBATE PEROXIDASE 1 do not accumulate NO or H2S following HS. Our findings shed new light on the intricate relationships between H2O2, NO, and H2S in plants, and pave the way for future studies of these three key signalling molecules.

Keywords: Hydrogen sulfide, Hydrogen peroxide, Imaging, Nitric oxide, Plant, Stress

Graphical abstract

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Highlights

  • •

    A live whole-plant imaging method was developed to study H2S signaling.

  • •

    The method could be used side-by-side with live imaging of NO and H2O2.

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    Intricate relationships between H2S, NO, and H2O2 were revealed during heat stress.

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    A mutant deficient in APX1 did not accumulate H2S or NO following heat stress.

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    A mutant deficient in RBOHD accumulated high basal levels of NO but not H2S.

1. Introduction

Hydrogen peroxide (H2O2), nitric oxide (NO), and hydrogen sulfide (H2S) are thought to regulate the structure, localization, and function of multiple proteins and enzymes in cells, thereby impact processes such as metabolic fluxes, growth, development, and abiotic/biotic stress responses [[1], [2], [3], [4], [5], [6]]. The levels of H2O2, that is highly soluble in aqueous media, are regulated by controlling the rates of H2O2 production [e.g., via enzymes such as respiratory burst oxidase homologs (RBOHs) coupled with superoxide dismutases (SODs)], scavenging [e.g., via enzymes such as catalases (CATs) and/or ascorbate peroxidases (APXs)], and transport [e.g., via peroxiporins/aquaporins (AQPs)] [7]. The levels of NO, a soluble molecule/gas, are mainly controlled by NO production [e.g., via nitrate reductase (NIA) and/or nitric oxide associated (NOA)], NO homeostasis (e.g., via GSNO reductase), and NO scavenging [e.g., via phytoglobins (PGBs)] [[8], [9], [10], [11]]. The levels of H2S, a soluble gaseous molecule, are controlled by regulating production [e.g., via l-cysteine desulfhydrases (DESs) in the cytosol, sulfite reductases (SIRs) in the chloroplast, and/or d-cysteine desulfhydrases (DCDs) in the mitochondria] and H2S scavenging [e.g., via O-acetylserine (thiol)lyases (OAS-TLs)/cysteine synthases (CSs)] [[12], [13], [14], [15]].

Hydrogen peroxide can impact protein structure and function by oxidizing cysteine and methionine residues via a posttranslational modification (PTM) process termed oxidative PTM (oxiPTM) and this process is mostly reversible via the function of glutathione peroxidases (GPXs), glutaredoxins (GRXs), thioredoxins (TRXs) and peroxiredoxin (PRXRs) [7,[16], [17], [18]]. Nitric oxide and NO-derived molecules can also mediate PTMs, mainly S-nitrosylation and tyrosine nitration, that are reversible (denitrosylation) via S-nitrosoglutathione reductases (GSNORs), TRXs, glutaredoxins (ROXYs or CC-type GRXs) and PRXRs [4,[19], [20], [21], [22]]. Hydrogen sulfide can cause an oxiPTM designated persulfidation, that can be reversed via NADPH/thioredoxin reductase (NTRB)/TRX1 and/or the glutathione/GRX system [23,24].

In addition to controlling multiple growth, developmental, and stress response processes in plants, H2O2, NO, and H2S are thought to impact the levels of each other via PTMs of different protein/enzymes. For example, NO and H2S were found to modulate the activity of RBOHD, a major source of O2−. (that is converted to H2O2 by SODs) in Arabidopsis, via S-nitrosylation and persulfidation, respectively. Thus, Arabidopsis thaliana RESPIRATORY BURST OXIDASE HOMOLOG D (AtRBOHD) is inhibited by S-nitrosylation Cys 890 [25] whereas it is stimulated by persulfidation of the same Cys 890 [26]. Interestingly, NO and H2S were also found to regulate the function of APXs and CATs via S-nitrosylation and persulfidation, respectively, impacting the levels of H2O2 scavenging. Thus, ASCORBATE PEROXIDASE 1 (APX1) is negatively regulated by tyrosine nitration but upregulated by S-nitrosylation and persulfidation [[27], [28], [29], [30]], whereas CAT is negatively regulated by tyrosine nitration, S-nitrosylation and persulfidation, mediated by NO and H2S, respectively [31,32]. While much is known about how NO and H2S can regulate H2O2 levels, the impact of H2O2 on NO and H2S levels in plant cells requires further studies. In mammals, H2O2 was found to enhance the activity of endothelial NO synthase (eNOS) via stimulating phosphorylation/dephosphorylation reactions [33,34], as well as to inhibit the activity of key H2S production enzymes such as cystathionine-lyase (CSE), cystathionine-synthase (CBS), and 3-mercaptopyruvate sulfurtransferase (MPST) [35,36]. Thus, at least in mammals and potentially also in plants [[37], [38], [39]], enhanced H2O2 levels have the ability to elevate NO production, while suppressing H2S levels.

The complex interactions among H2O2, NO, and H2S, highlighted above, should also be considered within the scope of the plant overall structure, as well as within the scope of plant-to-plant signaling. Plant structures include both cell-to-cell contacts [via cell wall (CW) and plasmodesmata (PD)] and airspaces. H2O2 can transverse cell-to-cell contacts, while NO and H2S can transverse cell-to-cell contacts as well as airspaces. NO and H2S can also transverse larger spaces, e.g., between leaves or root branches of the same plant, or even between different plants [40], while H2O2 can only be transferred between plants through physical touch [41]. H2O2, NO and H2S can also transverse intracellular structures and link the plasma membrane (PM) and organelle (e.g., chloroplast and mitochondria) signaling with the nuclei [40,[42], [43], [44]].

While multiple methods were developed for NO, H2O2, and H2S imaging at the cell and tissue levels (e.g., Ref. [[45], [46], [47], [48], [49]]), live methods to image the aboveground parts of whole plants grown in soil were only developed so far for H2O2 [50,51] and NO [52]. Here, we report on the development of a live whole-plant (aboveground) method for H2S imaging. This method is non-destructive, can be used on live plants grown in soil, is suitable for H2S detection in multiple wild type (WT) and mutant plants smaller than 20 cm, does not require plant transformation, and could be used in conjunction with different stress treatments and/or developmental growth phases. Using this method, we show that heat stress (HS), or treatment with the elicitor flg22, causes a whole plant accumulation of H2S, and that following HS occurring under low light conditions NO and H2S accumulation could be uncoupled from H2O2 accumulation. In addition, we reveal that while H2O2 accumulates in WT and mutants deficient in APX1 (apx1) or NIA1, NIA2 and NOA1 (nia1nia2noa1; deficient in NO synthesis) subjected to a HS treatment, H2S and NO do not accumulate in apx1 and nia1nia2noa1 plants under the same conditions. In addition, we show that NO, but not H2S, accumulates in rbohd mutants grown under controlled growth conditions. Our findings reveal that NO and H2S accumulation can be uncoupled from H2O2 accumulation following HS, depending on light stress intensity, that H2S accumulation following HS requires NO accumulation, and that APX1 is required for NO and H2S accumulation following HS.

2. Methods

2.1. Plant material and growth conditions

Arabidopsis thaliana plants cv. Columbia-0 wild-type (WT), rbohd (AT5G47910; [53]), apx1 (AT1G07890; [54]), nia1nia2noa1-2 [55] and des1-1 (AT5G28030; [56]) plants were grown on peat pellets (Jiffy 7; Jiffy International, Kristiansand, Norway) for 4-5 weeks under a 10 h/14 h; light (80 μmol photons s−1 m−2)/dark regime at 20 °C, 65% relative humidity. The light cycle started at 07:00 h and ended at 17:00 h. des1-1 T-DNA insertion mutant seeds were obtained from the Arabidopsis Biological Resource Center (ABRC; https://abrc.osu.edu/).

2.2. Application of fluorescent dyes, H2S donor, and/or H2S scavenger

Four to five-week-old Arabidopsis plants were fumigated using a nebulizer (Punasi Direct, Hong Kong, China) in a glass chamber as described by Refs. [[50], [51], [52],57], with: (1) 10 μM HSip-1 DA trifluoroacetate hydrate (HSip-1 DA; a cell permeable fluorescent H2S indicator, excitation/emission 491/516 nm; [58,59]; cat-1346170-03-3; Sigma-Aldrich, St. Louis, MO, USA); (2) 10 μM Sulfidefluor-7 acetoxymethyl ester (SF7-AM; fluorescent H2S indicator, excitation/emission 498/526 nm; [[59], [60], [61]]; cat-1416872-50-8; Cayman Chemical Co., Ann Arbor, MI, USA); (3) 10 μM WSP-1 (fluorescent H2S indicator, excitation/emission 465/515 nm; [46,62,63]; cat-1352750-34-5, MCE, Shanghai, China); (4) 100 μM GYY4137 Dichloromethane complex (GYY4137; H2S donor, [64]; cat-106740-09-4; Sigma-Aldrich, St. Louis, MO, USA); (5) 1 mM hypotaurine (H2S scavenger, [65]; cat-300-84-5; Sigma-Aldrich, St. Louis, MO, USA); (6) 15 μM 4,5-diaminofluorescein diacetate (DAF-2-DA; NO fluorescent indicator; excitation/emission 495 nm/517 nm; cat-205391-02-2; Chemodex, Gallen, Switzerland); and/or (7) 100 μM Peroxy Orange 1 (PO1; H2O2-specific dye; excitation/emission 540 nm/565 nm; [50]; cat-1199576-10-7; Tocris Bioscience, Bristol, UK). To perform fumigation all compounds were prepared in 10 mL of 10 mM phosphate buffer (pH 7.4) containing 0.002% Silwet L-77 (LEHLE seeds, Round Rock, TX, USA). To control for dye uptake into plants (e.g., in cases in which no signal was detected following plant heat stress treatment), at the end of the experiment plants were fumigated for 30 min with the compound the dye was meant to detect (H2O2, or NO or H2S donors) and immediately imaged again, as described by Fichman et al. [50].

For H2S scavenging experiments, plants were fumigated with phosphate buffer with or without hypotaurine for 30 min in separate glass chambers. Immediately upon completion of this first round of fumigation, buffer-treated plants were fumigated with HSip-1 DA and hypotaurine-treated plants were fumigated with HSip-1 DA + hypotaurine for an additional 30 min. It is extremely important to note that hypotaurine solution must be made fresh from powder right before fumigation to ensure maximum effectiveness. Likewise, for the H2S donor experiments, plants were fumigated with HSip-1 DA for 30 min and left inside the chamber for 30 min. Then, half of the plants were fumigated with GYY4137 + HSip-1 DA, and the other half were fumigated again with HSip-1 DA for an additional 30 min in separate chambers. For H2S imaging with WSP1, we used 10-day-old A. thaliana seedlings grown on ½ MS plates (Fig. S1), as this dye did not penetrate into mature plants.

2.3. Stress treatment

Post fumigation whole plant heat stress was performed by subjecting the entire plants, or agar plates, to 40 °C for 10 min under 5 or 100 photons s−1 m−2 light conditions [52]. In contrast, local heat stress was given by exposing a single leaf of the plant to a pre-heated heating block for 2 min under 100 photons s−1 m−2 light condition [66]. A thermal camera was used to ensure that the exposed leaf temperature experiences 40 °C. Flg22 treatment was applied by dipping two to three local leaves in 10 mM phosphate buffer (pH 7.4) containing 1 μM flg22 (PhytoTech Labs, Lenexa KS, USA), 0.002% Silwet L-77, and HSip-1 DA, whereas leaves dipped in buffer containing Silwet L-77 and HSip-1 DA served as mock treatment.

2.4. Image acquisition

Immediately after applying stress treatments to plants, plants were imaged using an IVIS Lumina S5 platform. Images were then processed using Living Image v4.7.3 (PerkinElmer) with fixed acquisition parameters. Fluorescence signals were acquired at 480/520 nm (HSip-1 DA, SF7-AM, WSP-1, and DAF-2-DA) or 540/565 nm (PO1) using f/2, 4-s exposure, and small binning. Images were acquired every 60 s for 30 min [50,52].

2.5. Image data and statistical analysis

The total radiant efficiency of regions of interest (ROIs) was measured using ImageJ as described by Ref. [52]. Each experiment was performed at least three times with four biological replicates. Data from all experiments was pooled to generate the final figure. Student's t-test was performed to evaluate the level of significance (p-value <0.05) in experiments that involve one genotype and one treatment. One-way ANOVA with Fisher's Least Significant Difference post-hoc test was performed to determine significant differences in graphs with one genotype and multiple treatments. Two-way ANOVA with Fisher's Least Significant Difference post-hoc test was performed to determine significant differences in graphs with multiple genotypes and multiple treatments. Different letters denote significant difference at p-value <0.05. As different treatments were used in different figures and these may require longer or shorter exposure times during imaging (due to differences in fold change of signal intensity), the baseline level of H2O2, H2S, or NO in untreated control plants cannot be compared between different figures.

3. Results

3.1. Developing a whole-plant live imaging method of H2S

Intracellular H2S can be detected using a diverse group of fluorescent and near-infrared fluorescent probes (Table S1), each based on a different mode of action ([[67], [68], [69]]; [70]; [46,[58], [59], [60], [61], [62], [63],71,72]). Detection of H2S by the fluorescent probe WSP-1 (Washington State Probe-1; [46,63]) is based on a two-step nucleophilic reaction sequence. H2S attacks the pyridyl disulfide group that leads to formation of a thiol intermediate and releases 2-thiopyridone, which then cyclizes to fluorescein, producing a strong fluorescence signal [70]. The fluorescent H2S probe SF7-AM (Sulfidefluor 7 a.m.; [[59], [60], [61]]) mode-of-function is based on a reaction between H2S and its azide groups. Following activation of the dye by intracellular esterases, H2S converts the azides to amines, restoring fluorophore conjugation which leads to an irreversible and selective fluorescence signal [67,69]. The fluorescent H2S probe HSip-1DA (Hydrogen Sulfide imaging probe-1 Diacetate; [58,59]), a metal-based probe, functions by Cu2+ displacement. Following activation of the dye by intracellular esterases, H2S binds to the central Cu2+ of the probe to form an insoluble CuS which relaxes the dye quenching and triggers fluorescence with high selectivity and sensitivity [68].

Intracellular H2S levels can also be chemically suppressed by hypotaurine that quenches H2S via an oxidation reaction that turns it into taurine [73]. In contrast, GYY4137 gradually and slowly releases H2S through hydrolysis of the phosphorodithioate bonds of GYY4137 followed by sulfur reduction which results in H2S release [74].

We previously developed methods for the whole-plant live imaging of H2O2, NO, and general reactive oxygen species (ROS) in Arabidopsis thaliana (A. thaliana) plants grown in soil [[50], [51], [52]]. These methods were based on applying different dyes via placing plants in a glass container for 30 min together with medical nebulizers that converted the dye (solvent in buffer) into a mist of small droplets that then entered the plant (confirmed via Z-stacking analysis performed with a confocal fluorescence microscope; [50]). Following fumigation of plants for 30 min, plants were treated/untreated and imaged for 30 to 60 min using an IVIS Lumina S5 fluorescent, white light, and bioluminescence imager [50]; originally developed to image fluorescent proteins under the skin of mice [75]. This optical high-sensitivity bioluminescence and fluorescence imaging system is based on a super cooled charge-coupled device (CCD) camera, and can image plants grown in soil or other media, such as agar or hydroponics, using an 20 × 20 cm imaging area (https://www.revvity.com/product/ivis-lumina-s5-imaging-system-cls148588; [[50], [51], [52]]). To image H2S using this setup, we first tested three different dyes, previously used to image H2S in cells: HSip-1 DA (Fig. 1A), WSP-1 (Fig. S1), and SF7-AM (Fig. S2). Of these, we chose to work with HSip-1 DA, as it provided us with the highest signal intensity and was permeable to mature plants.

Fig. 1.

Fig. 1

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Whole-plant live H2S imaging in the presence or absence of an H2S donor or scavenger. A. Representative time-lapse images (left) and a bar graph showing statistical analysis at 30 min (right) of whole-plant H2S imaging in wild type A. thaliana plants grown under controlled conditions and treated with the H2S donor GYY4137. Statistical analysis was performed with a Student's t-test, N ≥ 8; ∗p < 0.05. B. Same as in A, but for wild type A. thaliana plants grown under controlled conditions and subjected to heat stress (HS) in the presence or absence of the H2S scavenger hypotaurine. Statistical analysis was performed with one-way ANOVA followed by a Fisher LSD post hoc test, N ≥ 8; Different letters denote significance (p < 0.05). Borders of each box plot indicate the 25th and 75th percentiles of collected data, and each point is depicted within the plot. The internal horizontal line on each box plot corresponds to the median while the “X” corresponds to the mean. Whiskers represent the 1.5x interquartile data range. Dotted lines indicate the image areas (regions of interest) used to quantify signal intensity. Abbreviations: cm, centimeters; min, minutes.

To test the specificity of HSip-1 DA in mature A. thaliana plants, we used the H2S donor GYY4137 [Same concentration as used by Li et al. [74] and Pantaleno et al. [64], heat stress (HS) treatment [a known inducer of H2S accumulation; e.g., Ref. [76]; Li et al. [77]; same temperature and time as used by Mohanty et al. [52], and the H2S scavenger hypotaurine [Same concentration as used by Ortega et al. [73] and Shi et al. [65]. As shown in Fig. 1A, treatment of plants with the H2S donor GYY4137 prior to applying the HSip-1 DA dye resulted in enhanced HSip-1 DA fluorescence. In addition, subjecting plants to a 10 min HS treatment of 40oC resulted in an enhanced HSip-1 DA fluorescence signal, and this signal was quenched by co-treatment of plants with the H2S scavenger hypotaurine (Fig. 1B). To further test the specificity of the method we developed for H2S (Fig. 1), we used the l-CYSTEINE DESULFHYDRASE 1 (des1) mutant, deficient in the primary cytosolic producer of H2S in A. thaliana [42,56,78]. As shown in Fig. 2A, compared to wild type (WT) plants, the des1 mutant displayed significantly lower fluorescence of the HSip-1 DA dye following a HS treatment of 40oC for 10 min. As responses to pathogen infection are also accompanied with H2S accumulation (e.g., Ref. [78,79]), we further tested H2S accumulation in WT and the des1 mutant in response to treatment with the elicitor flg22, a highly conserved 22-amino acid peptide derived from the N-terminus of bacterial flagellin that acts as a potent pathogen-associated molecular pattern (PAMP) inducer [e.g., Ref. [80]; we used the same concentration of flg22 as reported by Mohanty et al. [52]. As shown in Fig. 2B, compared to WT plants that displayed enhanced fluorescence of HSip-1 DA in response to the flg22 treatment, des1 plants displayed significantly lower levels of HSip-1 DA fluorescence.

Fig. 2.

Fig. 2

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Hydrogen sulfide (H2S) accumulation is suppressed in the l-CYSTEINE DESULFHYDRASE 1 (des1) mutant. A. Representative time-lapse images (left) and a bar graph showing statistical analysis at 30 min (right) of whole-plant H2S imaging in wild type (WT) and des1 A. thaliana plants grown under controlled conditions or subjected to heat stress. B. Same as in A, but for WT and des1 mutants treated or untreated with flg22. Statistical analysis was performed with two-way ANOVA followed by a Fisher LSD post hoc test, N ≥ 8; Different letters denote significance (p < 0.05). Borders of each box plot indicate the 25th and 75th percentiles of collected data, and each point is depicted within the plot. The internal horizontal line on each box plot corresponds to the median while the “X” corresponds to the mean. Whiskers represent the 1.5x interquartile data range. Dotted lines indicate the image areas (regions of interest) used to quantify signal intensity. Abbreviations: cm, centimeters; des1, l-cysteine desulfhydrase 1; FLG22, flagellin 22; m, meters; min, minutes; s, seconds; WT, wild type.

Taken together, the findings presented in Figs. 1 and 2, S1, and S2, support the specificity and biological relevance of the method we developed for H2S imaging.

3.2. Imaging of H2S, NO, and H2O2 levels in A. thaliana plants subjected to HS under moderate or low light

H2S, NO, and H2O2 are thought to play a key role in regulating thermotolerance in plants (e.g., Ref. [52,76,81]). We previously reported that the levels of H2O2 and NO increased in A. thaliana in response to HS (40oC, 10 min) under moderate light (100 μmol photons s−1 m−2), but that only NO levels increased if the same HS treatment was conducted under low light conditions (5 μmol photons s−1 m−2; [52]). Thus, depending on light intensity, NO and H2O2 levels could be uncoupled during HS in A. thaliana. The dependency of H2O2 accumulation on light intensity during HS [52] allowed us to test whether H2S levels in cells are associated with H2O2 or NO during HS in A. thaliana. We hypothesized that in A. thaliana plants subjected to HS under low light, when H2O2 levels are suppressed and NO levels are elevated, H2S levels will be elevated. To test whether H2S accumulation in plants in response to HS is associated with H2S accumulation, we subjected WT A. thaliana plants to HS (40oC, 10 min) under low (5 μmol photons s−1 m−2), or moderate (100 μmol photons s−1 m−2), light conditions [52] and imaged H2S, NO, and H2O2. As shown in Fig. 3, under moderate light conditions, HS resulted in the accumulation of H2S, NO, and H2O2. In contrast, as shown in Fig. 4, under low light conditions, the same HS treatment resulted in the accumulation of H2S and NO, but not H2O2. The accumulation pattern of NO and H2O2 in response to HS under low light conditions agreed with Mohanty et al. [52], and could be the outcome of either light being needed for H2O2 accumulation during HS and/or reflect a suppressive mechanism of H2O2 accumulation by NO under low light. In contrast, the finding that H2S accumulated following HS occurring under low light conditions, when H2O2 levels are suppressed (Fig. 4), suggests that under these conditions H2S accumulation is not dependent on H2O2 levels, and could be associated with NO levels. Taken together, the findings presented in Fig. 3, Fig. 4 suggest that while H2O2 and NO or H2S accumulation can be uncoupled in response to HS, pending light intensity, H2S and NO accumulation cannot.

Fig. 3.

Fig. 3

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Whole-plant live H2S, NO and H2O2 imaging of wild type A. thaliana plants subjected to heat stress under moderate light. Representative time-lapse images (left) and a bar graph showing statistical analysis at 30 min (right) of whole-plant H2S (top), NO (middle), and H2O2 (bottom) imaging of A. thaliana plants grown under controlled conditions or subjected to heat stress under a light intensity of 100 μmol photons s−1 m−2 are shown. Statistical analysis was performed with a Student's t-test, N ≥ 8; ∗p < 0.05. Borders of each box plot indicate the 25th and 75th percentiles of collected data, and each point is depicted within the plot. The internal horizontal line on each box plot corresponds to the median while the “X” corresponds to the mean. Whiskers represent the 1.5x interquartile data range. Dotted lines indicate the image areas (regions of interest) used to quantify signal intensity. Abbreviations: cm, centimeters; H2O2, hydrogen peroxide; H2S, hydrogen sulfide; m, meters; min, minutes; NO, nitric oxide; s, seconds.

Fig. 4.

Fig. 4

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Whole-plant live H2S, NO and H2O2 imaging of wild type A. thaliana plants subjected to heat stress under low light. Representative time-lapse images (left) and a bar graph showing statistical analysis at 30 min (right) of whole-plant H2S (top), NO (middle), and H2O2 (bottom) imaging of A. thaliana plants grown under controlled conditions or subjected to heat stress under a light intensity of 5 μmol photons s−1 m−2 are shown. Statistical analysis was performed with a Student's t-test, N ≥ 8; ∗p < 0.05. Borders of each box plot indicate the 25th and 75th percentiles of collected data, and each point is depicted within the plot. The internal horizontal line on each box plot corresponds to the median while the “X” corresponds to the mean. Whiskers represent the 1.5x interquartile data range. Dotted lines indicate the image areas (regions of interest) used to quantify signal intensity. Abbreviations: cm, centimeters; H2O2, hydrogen peroxide; H2S, hydrogen sulfide; m, meters; min, minutes; NO, nitric oxide; s, seconds.

3.3. Imaging of H2S, NO, and H2O2 levels in WT, rbohd, apx1, and nia1nia2noa1 A. thaliana plants subjected to HS under moderate light conditions

To further study the relationship between H2O2, NO, and H2S accumulation during HS under moderate light, we subjected WT and different mutants impaired in H2O2 or NO levels to HS and imaged H2O2, NO, and H2S levels (Fig. 5, Fig. 6, Fig. 7). For this purpose, we used mutants deficient in H2O2 production (rbohd; [53,57,82]) and scavenging (apx1; [54,83]), as well as in NO accumulation (nia1nia2noa1; [52,55]). By using these mutants, we aimed to test several hypotheses. Using mutants deficient in H2O2 accumulation (rbohd) or scavenging (apx1) we wanted to test whether H2O2 is required for NO or H2S accumulation during HS under moderate light. Using a mutant deficient in NO accumulation (nia1nia2noa1), we wanted to test whether NO is required for H2S and/or H2O2 accumulation during HS under moderate light. As both RBOHD and APX1 are post-translationally regulated by NO and H2S [[25], [26], [27], [28], [29], [30]], we also wanted to test whether the levels of NO and H2S are altered in the rbohd and apx1 mutants under control conditions or in response to stress (as part of a feedback regulatory mechanism).

Fig. 5.

Fig. 5

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Whole-plant live H2O2 imaging of wild type, rbohd, apx1, and nia1nia2noa1 A. thaliana plants subjected to heat stress under moderate light. Representative time-lapse images (left) and a bar graph showing statistical analysis at 30 min (right) of whole-plant H2O2 imaging of wild type (WT), rbohd, apx1, and nia1nia2noa1 A. thaliana plants grown under controlled conditions or subjected to heat stress under a light intensity of 100 μmol photons s−1 m−2 are shown. Statistical analysis was performed with two-way ANOVA followed by a Fisher LSD post hoc test, N ≥ 8; different letters denote significance; p < 0.05. Borders of each box plot indicate the 25th and 75th percentiles of collected data, and each point is depicted within the plot. The internal horizontal line on each box plot corresponds to the median while the “X” corresponds to the mean. Whiskers represent the 1.5x interquartile data range. Dotted lines indicate the image areas (regions of interest) used to quantify signal intensity. Abbreviations: apx, ascorbate peroxidase; cm, centimeters; H2O2, hydrogen peroxide; m, meters; min, minutes; nia, nitrate reductase; noa, NO associated; rbohd, respiratory burst oxidase homolog D; s, seconds.

Fig. 6.

Fig. 6

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Whole-plant live NO imaging of wild type, rbohd, apx1, and nia1nia2noa1 A. thaliana plants subjected to heat stress under moderate light. Representative time-lapse images (left) and a bar graph showing statistical analysis at 30 min (right) of whole-plant NO imaging of wild type (WT), rbohd, apx1, and nia1nia2noa1 A. thaliana plants grown under controlled conditions or subjected to heat stress under a light intensity of 100 μmol photons s−1 m−2 are shown. Statistical analysis was performed with two-way ANOVA followed by a Fisher LSD post hoc test, N ≥ 8; different letters denote significance; p < 0.05. Borders of each box plot indicate the 25th and 75th percentiles of collected data, and each point is depicted within the plot. The internal horizontal line on each box plot corresponds to the median while the “X” corresponds to the mean. Whiskers represent the 1.5x interquartile data range. Dotted lines indicate the image areas (regions of interest) used to quantify signal intensity. Abbreviations: apx, ascorbate peroxidase; cm, centimeters; m, meters; min, minutes; nia, nitrate reductase; NO, nitric oxide; noa, NO associated; rbohd, respiratory burst oxidase homolog D; s, seconds.

Fig. 7.

Fig. 7

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Whole-plant live H2S imaging of wild type, rbohd, apx1, and nia1nia2noa1 A. thaliana plants subjected to heat stress under moderate light. Representative time-lapse images (left) and a bar graph showing statistical analysis at 30 min (right) of whole-plant H2S imaging of wild type (WT), rbohd, apx1, and nia1nia2noa1 A. thaliana plants grown under controlled conditions or subjected to heat stress under a light intensity of 100 μmol photons s−1 m−2 are shown. Statistical analysis was performed with two-way ANOVA followed by a Fisher LSD post hoc test, N ≥ 8; different letters denote significance; p < 0.05. Borders of each box plot indicate the 25th and 75th percentiles of collected data, and each point is depicted within the plot. The internal horizontal line on each box plot corresponds to the median while the “X” corresponds to the mean. Whiskers represent the 1.5x interquartile data range. Dotted lines indicate the image areas (regions of interest) used to quantify signal intensity. Abbreviations: apx, ascorbate peroxidase; cm, centimeters; H2S, hydrogen sulfide; m, meters; min, minutes; nia, nitrate reductase; noa, NO associated; rbohd, respiratory burst oxidase homolog D; s, seconds.

As shown in Fig. 5, under the growth conditions we used (moderate light), and in agreement with prior studies [54,57,82], none of three mutants accumulated high basal levels of H2O2 (Fig. 5). In response to the whole plant HS treatment, WT, as well as all 3 mutants, accumulated H2O2 (Fig. 5). While this finding was expected for WT, apx1 and nia1nia2noa1, it was surprising that rbohd accumulated H2O2, as this mutant was previously reported to be deficient in whole-plant ROS accumulation (imaged with 2′,7′-dichlorofluorescein, DCF) following a local treatment of HS ([84]; confirmed here again Fig. S3). It is therefore possible that RBOHD plays different roles in plants subjected to local or whole-plant HS treatments. The results shown in Fig. 5 suggest that H2O2 accumulation during HS under moderate light is not dependent on NO accumulation in WT or the different mutant backgrounds studied.

When WT, rbohd, apx1, and nia1nia2noa1 plants were subjected to the same HS treatment (whole plant HS of 40oC for 10 min under 100 μmol photons s−1 m−2) and imaged for NO accumulation (Fig. 6), it was found that NO accumulated in WT in response to the HS treatment, but not in apx1 and nia1nia2noa1 plants. While this result was expected for the nia1nia2noa1 triple mutant that is deficient in NO accumulation [52,55], the finding that NO levels did not accumulate in apx1 plants following HS was surprising. In addition, as shown in Fig. 6, compared to WT grown under controlled conditions, rbohd plants accumulated significantly higher levels of NO under controlled conditions or following HS. The altered levels of NO in the rbohd (elevated under controlled growth conditions, compared to WT) and apx1 (deficient following HS under moderate light, compared to WT) mutants (Fig. 6) could result from a feedback regulatory mechanism controlling NO levels in these mutants.

When WT, rbohd, apx1, and nia1nia2noa1 plants were subjected to the same HS treatment and imaged for H2S accumulation (Fig. 7), it was found that H2S accumulated in WT and rbohd plants in response to HS, but not in apx1 and nia1nia2noa1 plants. This result revealed that, at least in WT plants, in response to HS, H2S and NO accumulation could not be uncoupled. In addition, as shown in Fig. 7, rbohd plants accumulated significantly higher levels of H2S compared to WT in response to HS, but did not contain high levels of H2S when grown under controlled growth conditions (rbohd plants grown under controlled growth conditions were found to have high basal levels of NO; Fig. 6). Thus, at least in rbohd plants grown under controlled growth conditions the levels of NO and H2S could be uncoupled (Fig. 6, Fig. 7). The results shown in Fig. 7 could suggest that H2S accumulation during HS under moderate light is dependent on NO accumulation.

Taken together, the results presented in Fig. 5, Fig. 6, Fig. 7 suggest a complex relationship between H2O2, NO, and H2S during plant responses to HS.

4. Discussion

We developed a method for whole plant imaging of H2S that complements the H2O2 [50,51] and NO [52] methods we previously developed and could be used side-by-side with them (Fig. 3, Fig. 4, Fig. 5, Fig. 6, Fig. 7). We confirmed the specificity and biological relevance of our method using an H2S donor (Fig. 1A), an H2S scavenger (Fig. 1B), and the des1 mutant (Fig. 2), as well as treatment of plants with HS under low or moderate light (Figs. 1B-7) or flg22 (Fig. 2). Using side-by-side imaging of H2O2, NO and H2S, following HS, we showed that in WT plants, under low light conditions, H2O2 accumulation could be uncoupled from H2S and NO accumulation (Fig. 3, Fig. 4). The implications of this finding are important, as H2O2, NO and H2S were proposed to regulate the levels of each other [5,37,85,86]. Under low light, it is possible therefore that H2O2 is not needed to promote NO and H2S accumulation (Fig. 4). In support of this uncoupling, we also found that H2O2 accumulation in the apx1 or nia1nia2noa1 mutants did not result in NO or H2S accumulation (Fig. 6, Fig. 7). Moreover, H2O2 accumulated in response to HS in the absence of H2S or NO in the apx1 or nia1nia2noa1 mutants (Fig. 5), suggesting that NO or H2S are not required for H2O2 accumulation under these conditions. Thus, at least in A. thaliana, WT, apx1 and nia1nia2noa1 mutants, following a HS treatment under moderate light, H2O2 accumulation did not cause NO or H2S accumulation and was not dependent on NO or H2S to accumulate (Fig. 5, Fig. 6, Fig. 7). As H2S levels in the des1 mutant were only suppressed and not completely blocked in response to HS (Fig. 2), we unfortunately could not use this mutant in the experiments shown in Fig. 5, Fig. 6, Fig. 7.

In contrast to the uncoupling between H2O2 and NO or H2S accumulation (Fig. 3, Fig. 4, Fig. 5, Fig. 6, Fig. 7), the levels of NO and H2S appeared to correspond with each other under most conditions we studied (Fig. 3, Fig. 4, Fig. 5, Fig. 6, Fig. 7; except in rbohd plants grown under controlled growth conditions). Thus, while H2O2 and H2S or NO could be uncoupled under conditions of HS occurring at low light, NO and H2S could not. In addition, following HS, the nia1nia2noa1 mutant that did not accumulate NO (Fig. 6; [52,55]), also did not accumulate H2S (Fig. 7), further supporting the tight relationship between NO and H2S levels in cells. Further studies are needed to decipher the relationships between NO and H2S levels in plants, especially since NO may be needed for H2S accumulation (Fig. 6, Fig. 7), and both molecules have a prominent effect on the structure and function of multiple proteins in cells [25,26,29,87,88]. Although it was previously proposed that H2S functions downstream of NO during heat stress in maize seedlings ([81]; supported by our findings that H2S did not accumulate in the nia1nia2noa1 triple mutant that does not accumulate NO during heat stress; Fig. 7), we could not detect any temporal differences between the accumulation of these two redox regulators in wild type plants. In future studies we plan to conduct more detailed time-course experiments for H2S, NO, and H2O2 accumulation during heat stress in Arabidopsis.

The findings that H2O2 levels did not accumulate in response to HS under low light conditions (Fig. 5), nor were they dependent on RBOHD under moderate light (Fig. 5), suggest that H2O2 generated during the whole-plant HS treatment, in our experimental setup, were produced in chloroplasts (in which H2O2 production is mostly dependent on light; [7,89].). If NO and H2S, generated in cells during HS, were produced outside the chloroplast (e.g., in the mitochondria or cytosol), the levels of NO and H2S might have less impact on the levels of chloroplastic-generated H2O2 (as they were not produced in the same cellular compartment; albeit, they should be able to travel between organelles). As our study suggests that, in WT plants, NO and H2S levels could be uncoupled from H2O2, further studies are needed to identify the subcellular sources of H2O2, NO and H2S under the controlled and HS conditions we used, as well as other abiotic/biotic stresses. Such studies should unravel how H2S, NO, and H2O2 levels are coordinated, and/or regulate each other, between the different subcellular compartments of plants.

Somewhat of a mystery is our finding that NO or H2S did not accumulate in the apx1 mutant following HS (Fig. 6, Fig. 7). As the apx1 mutant was found to uptake the dye we used for H2O2 imaging (Fig. 5), it is unlikely that it would not uptake the dyes we used for NO or H2S imaging due to enhanced stomatal conductance. ASCORBATE PEROXIDASE 1 is thought to regulate the levels of H2O2 in the cytosol and mutants deficient in APX1 are more susceptible to excess light stress [54,83]. In addition, these mutants are smaller in size and undergo different metabolic and molecular alterations [54,83,90,91]. It is therefore possible that in these mutants the expression and/or activity of the different enzymes required for NO and H2S production are suppressed. Further studies of the apx1 mutant are needed to resolve this mystery, especially since APX1 is regulated by both NO and H2S [27,91] and the absence of their accumulation in the apx1 mutant could reflect a feedback regulatory mechanism between APX1 protein levels and NO and/or H2S signaling.

In addition to APX1, RBOHD is also regulated by NO and H2S [25,26]. Interestingly, in contrast to the apx1 mutant (Fig. 6, Fig. 7), we found that in the rbohd mutant NO levels were elevated under control conditions (Fig. 6), and H2S levels were higher than WT following the HS treatment (Fig. 7). As RBOHD was found to be inhibited by S-nitrosylation, or stimulated by persulfidation, of Cys 890 [25,26], the altered accumulation of NO and H2S in the rbohd mutant (Fig. 6, Fig. 7), could also reflect a feedback regulatory mechanism (between RBOHD and NO and/or H2S).

While the use of biosensors for H2O2 and redox has been successfully established in plants (e.g., Ref. [49,92]), to the best of our knowledge, similar protein-based reporters for NO and H2S in plants have not been developed yet. Thus, fluorescent dyes are still the best alternative for the in vivo imaging of these molecules in plants. Moreover, even if biosensors were developed for H2S and NO imaging in plants, they would need to be transformed/crossed into each mutant studied and their levels in WT and mutants studied standardized. The method presented here provides therefore a fast and efficient way to study H2S levels in plants (WT and different mutants), as well as to compare it to NO and H2O2 levels.

In future studies the different H2S, NO and H2O2 imaging methods described in this study could be used to test multiple other hypotheses regarding the roles of these important redox-signaling molecules. For example, H2O2 or donors of NO or H2S could be applied to WT plants and different mutants, grown under controlled conditions, or subjected to different stresses, followed by measurements of H2O2, NO, and/or H2S, to further study the dependency of these different compounds on each other. Similar experiments could be performed using biotic (e.g., flg22 or DC3000 treatments) and/or abiotic (e.g., excess light or cold stresses) stimuli. Finally, the role of NO, H2S, and H2O2 signaling could be studied in plants during different biotic and/or abiotic stress combinations. Similarly, gain- or loss-of-function mutants, altered in developmental processes or stress responses, could be phenotypically analyzed for H2S, NO and H2O2 levels in terms of complex redox regulation.

5. Conclusion

We developed a whole-plant live imaging method for H2S, validated it, and used it to study the intricate relationships between H2O2, NO, and H2S following HS in Arabidopsis thaliana. Our findings revealed that H2O2 accumulation could be uncoupled from H2S and NO accumulation following HS, but that H2S and NO accumulation are mostly tied to each other. We further show that H2S accumulation following HS requires NO accumulation, that rbohd mutants grown under controlled growth conditions accumulate high levels of NO, and that apx1 mutants do not accumulate NO or H2S following HS. Our study uncovers the complex relationships between H2S, NO and H2O2 signaling, and paves the way for future studies on these important redox regulators.

Funding

This work was supported by funding from the National Science Foundation (IOS-2414183; IOS-2110017; IOS-2343815) and the Interdisciplinary Plant Group, and University of Missouri, Columbia, and from the Spanish Ministry of Science, Innovation and Universities MCIU/AEI/10.13039/501100011033 (PID2023-149447OB-I00); Regional Government of Castile and Leon (SA142P23) and ‘Escalera de Excelencia’ CLU-2O25-2-04 co-funded by the FEDER Operative Program of Castile and Leon 2021–2027 Spain (to OL).

CRediT authorship contribution statement

Devasantosh Mohanty: Data curation, Formal analysis, Investigation, Methodology, Validation, Writing – review & editing. María Ángeles Peláez-Vico: Formal analysis, Investigation, Methodology, Validation, Writing – review & editing. Ranjita Sinha: Formal analysis, Investigation, Validation, Writing – review & editing. María Inmaculada Sánchez-Vicente: Data curation, Formal analysis, Investigation, Resources, Writing – review & editing. Oscar Lorenzo: Conceptualization, Investigation, Resources, Supervision, Writing – review & editing. Ron Mittler: Conceptualization, Funding acquisition, Investigation, Project administration, Supervision, Writing – original draft, Writing – review & editing.

Declaration of competing interest

The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

Acknowledgments

We thank the Arabidopsis Biological Resource Center (ABRC) for seeds of wild type and mutant A. thaliana plants.

Footnotes

Appendix A

Supplementary data to this article can be found online at https://doi.org/10.1016/j.redox.2026.104205.

Appendix A. Supplementary data

The following are the Supplementary data to this article:

Multimedia component 1
mmc1.pdf (239.2KB, pdf)
Multimedia component 2
mmc2.pdf (485.2KB, pdf)

Data availability

Data will be made available on request.

References

  • 1.Sánchez-Vicente I., Albertos P., Lorenzo O. Protein shuttle between nucleus and cytoplasm: new paradigms in the ABI5-dependent ABA responses. Mol. Plant. 2019;12:1425–1427. doi: 10.1016/j.molp.2019.09.005. [DOI] [PubMed] [Google Scholar]
  • 2.Corpas F.J., González-Gordo S., Cañas A., Palma J.M. Nitric oxide and hydrogen sulfide in plants: which comes first? J. Exp. Bot. 2019;70:4391–4404. doi: 10.1093/jxb/erz031. [DOI] [PubMed] [Google Scholar]
  • 3.Corpas F.J., Rivero R.M., Freschi L., Palma J.M. Functional interactions among H2O2, NO, H2S, and melatonin in the physiology, metabolism, and quality of horticultural solanaceae. J. Exp. Bot. 2025;76:3634–3656. doi: 10.1093/jxb/erae513. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 4.Corpas F.J., Taboada J., Palma J.M. Integration of NO and H2S signaling in higher plants. Plant Commun. 2026;7 doi: 10.1016/j.xplc.2025.101664. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 5.Mukherjee S., Corpas F.J. H2O2, NO, and H2S networks during root development and signalling under physiological and challenging environments: beneficial or toxic? Plant. Cell Environ. 2023;46:688–717. doi: 10.1111/pce.14531. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 6.Rai K.K., Corpas F.J. Interplay between transcription factors and redox-related genes in ROS, RNS and H2S signalling during plant stress responses. Plant Cell Environ. 2025;48:7796–7814. doi: 10.1111/pce.70077. [DOI] [PubMed] [Google Scholar]
  • 7.Mittler R., Zandalinas S.I., Fichman Y., Van Breusegem F. Reactive oxygen species signalling in plant stress responses. Nat. Rev. Mol. Cell Biol. 2022;23:663–679. doi: 10.1038/s41580-022-00499-2. [DOI] [PubMed] [Google Scholar]
  • 8.Lee U., Wie C., Fernandez B.O., Feelisch M., Vierling E. Modulation of nitrosative stress by S-nitrosoglutathione reductase is critical for thermotolerance and plant growth in arabidopsis. Plant Cell. 2008;20:786–802. doi: 10.1105/tpc.107.052647. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 9.Berger A., Boscari A., Puppo A., Brouquisse R. Nitrate reductases and hemoglobins control nitrogen-fixing symbiosis by regulating nitric oxide accumulation. J. Exp. Bot. 2021;72:873–884. doi: 10.1093/jxb/eraa403. [DOI] [PubMed] [Google Scholar]
  • 10.Terrón-Camero L.C., Molina-Moya E., Peláez-Vico M.Á., Sandalio L.M., Romero-Puertas M.C. Nitric oxide and globin Glb1 regulate Fusarium oxysporum infection of Arabidopsis thaliana. Antioxidants. 2023;12:1321. doi: 10.3390/antiox12071321. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 11.Pathak P.K., Yadav N., Kaladhar V.C., Jaiswal R., Kumari A., Igamberdiev A.U., Loake G.J., Gupta K.J. The emerging roles of nitric oxide and its associated scavengers-phytoglobins-in plant symbiotic interactions. J. Exp. Bot. 2024;75:563–577. doi: 10.1093/jxb/erad399. [DOI] [PubMed] [Google Scholar]
  • 12.Romero L.C., García I., Gotor C. L-cysteine desulfhydrase 1 modulates the generation of the signaling molecule sulfide in plant cytosol. Plant Signal. Behav. 2013;8 doi: 10.4161/psb.24007. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 13.Yarmolinsky D., Brychkova G., Fluhr R., Sagi M. Sulfite reductase protects plants against sulfite toxicity. Plant Physiol. 2013;161:725–743. doi: 10.1104/pp.112.207712. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 14.Aroca A., Gotor C., Romero L.C. Hydrogen sulfide signaling in plants: emerging roles of protein persulfidation. Front. Plant Sci. 2018;9:1369. doi: 10.3389/fpls.2018.01369. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 15.Fang H., Chen W., Xing K., Xu L., Wu J., Huang Y., Pei Y., Zang Y. Hydrogen sulfide generated by L-cysteine desulfhydrase 1 enhances plant basal thermotolerance through the regulation of stomatal behavior and the promotion of photosynthetic efficiency. Plant Sci. 2025;361 doi: 10.1016/j.plantsci.2025.112805. [DOI] [PubMed] [Google Scholar]
  • 16.Sevilla F., Camejo D., Ortiz-Espín A., Calderón A., Lázaro J.J., Jiménez A. The thioredoxin/peroxiredoxin/sulfiredoxin system: current overview on its redox function in plants and regulation by reactive oxygen and nitrogen species. J. Exp. Bot. 2015;66:2945–2955. doi: 10.1093/jxb/erv146. [DOI] [PubMed] [Google Scholar]
  • 17.Waszczak C., Akter S., Jacques S., Huang J., Messens J., Van Breusegem F. Oxidative post-translational modifications of cysteine residues in plant signal transduction. J. Exp. Bot. 2015;66:2923–2934. doi: 10.1093/jxb/erv084. [DOI] [PubMed] [Google Scholar]
  • 18.Jiménez A., Martí M.C., Sevilla F. Oxidative post-translational modifications of plant antioxidant systems under environmental stress. Physiol. Plantarum. 2025;177 doi: 10.1111/ppl.70118. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 19.Kneeshaw S., Gelineau S., Tada Y., Loake G.J., Spoel S.H. Selective protein denitrosylation activity of thioredoxin-h5 modulates plant immunity. Mol. Cell. 2014;6:153–162. doi: 10.1016/j.molcel.2014.08.003. [DOI] [PubMed] [Google Scholar]
  • 20.Tichá T., Lochman J., Činčalová L., Luhová L., Petřivalský M. Redox regulation of plant S-nitrosoglutathione reductase activity through post-translational modifications of cysteine residues. Biochem. Biophys. Res. Commun. 2017;494:27–33. doi: 10.1016/j.bbrc.2017.10.090. [DOI] [PubMed] [Google Scholar]
  • 21.León J. Protein tyrosine nitration in plant nitric oxide signaling. Front. Plant Sci. 2022;13 doi: 10.3389/fpls.2022.859374. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 22.Sánchez-Vicente I., Albertos P., Sanz C., Wybouw B., De Rybel B., Begara-Morales J.C., Chaki M., Mata-Pérez C., Barroso J.B., Lorenzo O. Reversible S-nitrosylation of bZIP67 by peroxiredoxin IIE activity and nitro-fatty acids regulates the plant lipid profile. Cell Rep. 2024;43 doi: 10.1016/j.celrep.2024.114091. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 23.De Brasi-Velasco S., Aroca A., Romero L.C., Gotor C., Sevilla F., Jiménez A. New role for thioredoxins in plants: implication of TRXo1 in protein depersulfidation. Redox Biol. 2025;82 doi: 10.1016/j.redox.2025.103627. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 24.Romero L.C., Carrillo R., Montesinos-Pereira D., Luque C., Aroca A., Gotor C. Protein persulfidation: a ubiquitous modification regulating a broad spectrum of biological processes. J. Exp. Bot. 2025 doi: 10.1093/jxb/eraf464. eraf464. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 25.Yun B.-W., Feechan A., Yin M., Saidi N.B.B., Le Bihan T., Yu M., Moore J.W., Kang J.-G., Kwon E., Spoel S.H., Pallas J.A., Loake G.J. S-nitrosylation of NADPH oxidase regulates cell death in plant immunity. Nature. 2011;478:264–268. doi: 10.1038/nature10427. [DOI] [PubMed] [Google Scholar]
  • 26.Shen J., Zhang J., Zhou M., Zhou H., Cui B., Gotor C., Romero L.C., Fu L., Yang J., Foyer C.H., Pan Q., Shen W., Xie Y. Persulfidation-based modification of cysteine desulfhydrase and the NADPH oxidase RBOHD controls guard cell abscisic acid signaling. Plant Cell. 2020;32:1000–1017. doi: 10.1105/tpc.19.00826. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 27.Begara-Morales J.C., Sánchez-Calvo B., Chaki M., Valderrama R., Mata-Pérez C., López-Jaramillo J., Padilla M.N., Carreras A., Corpas F.J., Barroso J.B. Dual regulation of cytosolic ascorbate peroxidase (APX) by tyrosine nitration and S-nitrosylation. J. Exp. Bot. 2014;65:527–538. doi: 10.1093/jxb/ert396. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 28.Aroca Á., Serna A., Gotor C., Romero L.C. S-sulfhydration: a cysteine posttranslational modification in plant systems. Plant Physiol. 2015;168:334–342. doi: 10.1104/pp.15.00009. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 29.Yang H., Mu J., Chen L., Feng J., Hu J., Li L., Zhou J.-M., Zuo J. S-nitrosylation positively regulates ascorbate peroxidase activity during plant stress responses. Plant Physiol. 2015;167:1604–1615. doi: 10.1104/pp.114.255216. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 30.Kolbert Z., Molnár Á., Oláh D., Feigl G., Horváth E., Erdei L., Ördög A., Rudolf E., Barth T., Lindermayr C. S-nitrosothiol signaling is involved in regulating hydrogen peroxide metabolism of zinc-stressed arabidopsis. Plant Cell Physiol. 2019;60:2449–2463. doi: 10.1093/pcp/pcz138. [DOI] [PubMed] [Google Scholar]
  • 31.Li J., Shi C., Wang Xiaofeng, Liu C., Ding X., Ma P., Wang Xiao, Jia H. Hydrogen sulfide regulates the activity of antioxidant enzymes through persulfidation and improves the resistance of tomato seedling to copper oxide nanoparticles (cuO NPs)-induced oxidative stress. Plant Physiol. Biochem. 2020;156:257–266. doi: 10.1016/j.plaphy.2020.09.020. [DOI] [PubMed] [Google Scholar]
  • 32.Palma J.M., Mateos R.M., López-Jaramillo J., Rodríguez-Ruiz M., González-Gordo S., Lechuga-Sancho A.M., Corpas F.J. Plant catalases as NO and H2S targets. Redox Biol. 2020;34 doi: 10.1016/j.redox.2020.101525. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 33.Thomas S.R., Chen K., Keaney J.F. Hydrogen peroxide activates endothelial nitric-oxide synthase through coordinated phosphorylation and dephosphorylation via a phosphoinositide 3-kinase-dependent signaling pathway. J. Biol. Chem. 2002;277:6017–6024. doi: 10.1074/jbc.M109107200. [DOI] [PubMed] [Google Scholar]
  • 34.Sartoretto J.L., Kalwa H., Pluth M.D., Lippard S.J., Michel T. Hydrogen peroxide differentially modulates cardiac myocyte nitric oxide synthesis. Proc. Natl. Acad. Sci. U. S. A. 2011;108:15792–15797. doi: 10.1073/pnas.1111331108. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 35.Wang M., Cao X., Luan C., Li Z. Hydrogen sulfide attenuates hydrogen peroxide-induced injury in human lung epithelial A549 cells. Int. J. Mol. Sci. 2019;20:3975. doi: 10.3390/ijms20163975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 36.Wang J., Xie H., Li H., Wang R., Zhang B., Ren T., Hua J., Chen N. NIR fluorescent probe for in situ bioimaging of endogenous H2S in rice roots under Al3+ and flooding stresses. J. Agric. Food Chem. 2021;69:14330–14339. doi: 10.1021/acs.jafc.1c05247. [DOI] [PubMed] [Google Scholar]
  • 37.Hancock J.T. Harnessing evolutionary toxins for signaling: reactive oxygen species, nitric oxide and hydrogen sulfide in plant cell regulation. Front. Plant Sci. 2017;8:189. doi: 10.3389/fpls.2017.00189. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 38.Liu H., Wang J., Liu J., Liu T., Xue S. Hydrogen sulfide (H2S) signaling in plant development and stress responses. aBIOTECH. 2021;2:32–63. doi: 10.1007/s42994-021-00035-4. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 39.Taziun T., Kumari R., Zaid A., Farooq B. Nitric oxide and hydrogen peroxide crosstalk enhances photosynthesis and nitrogen assimilation in tomato and brinjal under Cr (VI) stress. Discov. Environ. 2025;3:260. doi: 10.1007/s44274-025-00422-4. [DOI] [Google Scholar]
  • 40.Kolbert Z., Barroso J.B., Boscari A., Corpas F.J., Gupta K.J., Hancock J.T., Lindermayr C., Palma J.M., Petřivalský M., Wendehenne D., Loake G.J. Interorgan, intraorgan and interplant communication mediated by nitric oxide and related species. New Phytol. 2024;244:786–797. doi: 10.1111/nph.20085. [DOI] [PubMed] [Google Scholar]
  • 41.Peláez-Vico M.Á., Fichman Y., Ghani A., Dhakal S., Bereimipour A., Azad R., Karpiński S.M., Joshi T., Mittler R. United we stand: plants that physically touch each other are more resilient to excess light stress. 2025. [DOI]
  • 42.Álvarez C., García I., Moreno I., Pérez-Pérez M.E., Crespo J.L., Romero L.C., Gotor C. Cysteine-generated sulfide in the cytosol negatively regulates autophagy and modulates the transcriptional profile in Arabidopsis. Plant Cell. 2012;24:4621–4634. doi: 10.1105/tpc.112.105403. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 43.Exposito-Rodriguez M., Laissue P.P., Yvon-Durocher G., Smirnoff N., Mullineaux P.M. Photosynthesis-dependent H2O2 transfer from chloroplasts to nuclei provides a high-light signalling mechanism. Nat. Commun. 2017;8:49. doi: 10.1038/s41467-017-00074-w. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 44.Smirnoff N., Arnaud D. Hydrogen peroxide metabolism and functions in plants. New Phytol.t. 2019;221:1197–1214. doi: 10.1111/nph.15488. [DOI] [PubMed] [Google Scholar]
  • 45.Belousov V.V., Fradkov A.F., Lukyanov K.A., Staroverov D.B., Shakhbazov K.S., Terskikh A.V., Lukyanov S. Genetically encoded fluorescent indicator for intracellular hydrogen peroxide. Nat. Methods. 2006;3:281–286. doi: 10.1038/nmeth866. [DOI] [PubMed] [Google Scholar]
  • 46.Li Y.-J., Chen J., Xian M., Zhou L.-G., Han F.X., Gan L.-J., Shi Z.-Q. In site bioimaging of hydrogen sulfide uncovers its pivotal role in regulating nitric oxide-induced lateral root formation. PLoS One. 2014;9 doi: 10.1371/journal.pone.0090340. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 47.Zhang P., Luo Q., Wang R., Xu J. Hydrogen sulfide toxicity inhibits primary root growth through the ROS-NO pathway. Sci. Rep. 2017;7:868. doi: 10.1038/s41598-017-01046-2. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 48.Wang Jun, Jia G., Li H., Yan S., Qian J., Guo X., Li G., Qi H., Zhu Z., Wu Y., He W., Niu W. H2O2-mediated oxidative stress enhances cystathionine γ-lyase-derived H2S synthesis via a sulfenic acid intermediate. Antioxidants. 2021;10:1488. doi: 10.3390/antiox10091488. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 49.Ugalde J.M., Fecker L., Schwarzländer M., Müller-Schüssele S.J., Meyer A.J. In: Reactive Oxygen Species in Plants: Methods and Protocols. Mhamdi A., editor. Springer US; New York, NY: 2022. Live monitoring of ROS-induced cytosolic redox changes with roGFP2-based sensors in plants; pp. 65–85. [DOI] [PubMed] [Google Scholar]
  • 50.Fichman Y., Miller G., Mittler R. Whole-plant live imaging of reactive oxygen species. Mol. Plant. 2019;12:1203–1210. doi: 10.1016/j.molp.2019.06.003. [DOI] [PubMed] [Google Scholar]
  • 51.Fichman Y., Zandalinas S.I., Peck S., Luan S., Mittler R. HPCA1 is required for systemic reactive oxygen species and calcium cell-to-cell signaling and plant acclimation to stress. Plant Cell. 2022;34:4453–4471. doi: 10.1093/plcell/koac241. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 52.Mohanty D., Peláez-Vico M.Á., Myers Jr R.J., Sánchez-Vicente M.I., Lorenzo O., Mittler R. Aboveground whole-plant live imaging method for nitric oxide (NO) reveals an intricate relationship between NO and H2O2. New Phytol. 2025;247:2473–2483. doi: 10.1111/nph.70094. [DOI] [PubMed] [Google Scholar]
  • 53.Torres M.A., Dangl J.L., Jones J.D.G. Arabidopsis gp91phox homologues AtrbohD and AtrbohF are required for accumulation of reactive oxygen intermediates in the plant defense response. Proc. Natl. Acad. Sci. U. S. A. 2002;99:517–522. doi: 10.1073/pnas.012452499. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 54.Davletova S., Rizhsky L., Liang H., Shengqiang Z., Oliver D.J., Coutu J., Shulaev V., Schlauch K., Mittler R. Cytosolic ascorbate peroxidase 1 is a central component of the reactive oxygen gene network of arabidopsis. Plant Cell. 2005;17:268–281. doi: 10.1105/tpc.104.026971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 55.Lozano-Juste J., León J. Enhanced abscisic acid-mediated responses in nia1nia2noa1-2 triple mutant impaired in NIA/NR- and AtNOA1-dependent nitric oxide biosynthesis in Arabidopsis. Plant Physiol. 2010;152:891–903. doi: 10.1104/pp.109.148023. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 56.Baudouin E., Poilevey A., Hewage N.I., Cochet F., Puyaubert J., Bailly C. The significance of hydrogen sulfide for Arabidopsis seed germination. Front. Plant Sci. 2016;7:930. doi: 10.3389/fpls.2016.00930. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 57.Mohanty D., Fichman Y., Peláez-Vico M.Á., Myers R.J., Sealander M., Sinha R., Morrow J., Eckstein R., Olson K., Xu C., An H., Yoo C.Y., Zhu J.-K., Zhao C., Zandalinas S.I., Liscum E., Mittler R. Identification of a putative RBOHD-FERONIA-CRK10-PIP2;6 plasma membrane complex that interacts with phyB to regulate ROS production in Arabidopsis thaliana. 2025. [DOI] [PubMed]
  • 58.Fukudome M., Shimada H., Uchi N., Osuki K., Ishizaki H., Murakami E., Kawaguchi M., Uchiumi T. Reactive sulfur species interact with other signal molecules in root nodule symbiosis in Lotus japonicus. Antioxidants. 2020;9:145. doi: 10.3390/antiox9020145. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 59.Minguillón S., Matamoros M.A., Duanmu D., Becana M. Signaling by reactive molecules and antioxidants in legume nodules. New Phytol. 2022;236:815–832. doi: 10.1111/nph.18434. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 60.Zou H., Zhang N.-N., Pan Q., Zhang J.-H., Chen J., Wei G.-H. Hydrogen sulfide promotes nodulation and nitrogen fixation in soybean-rhizobia symbiotic system. MPMI (Mol. Plant-Microbe Interact.) 2019;32:972–985. doi: 10.1094/MPMI-01-19-0003-R. [DOI] [PubMed] [Google Scholar]
  • 61.Zou H., Zhang N.-N., Lin X.-Y., Zhang W.-Q., Zhang J.-H., Chen J., Wei G.-H. Hydrogen sulfide is a crucial element of the antioxidant defense system in Glycine max-Sinorhizobium fredii symbiotic root nodules. Plant Soil. 2020;449:209–231. doi: 10.1007/s11104-020-04465-9. [DOI] [Google Scholar]
  • 62.Devarie-Baez N.O., Bagdon P.E., Peng B., Zhao Y., Park C.-M., Xian M. Light-induced hydrogen sulfide release from “Caged” gem-dithiols. Org. Lett. 2013;15:2786–2789. doi: 10.1021/ol401118. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 63.Ye Q., Ren M., Fan D., Mao Y., Zhu Y.-Z. Identification and validation of the miR/RAS/RUNX2 autophagy regulatory network in AngII-induced hypertensive nephropathy in MPC5 cells treated with hydrogen sulfide donors. Antioxidants. 2024;13:958. doi: 10.3390/antiox13080958. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 64.Pantaleno R., Scuffi D., Schiel P., Schwarzländer M., Costa A., García-Mata C. Mitochondrial ß-cyanoalanine synthase participates in flg22-induced stomatal immunity. Plant Cell Environ. 2025;48:537–552. doi: 10.1111/pce.15155. [DOI] [PubMed] [Google Scholar]
  • 65.Shi H., Ye T., Han N., Bian H., Liu X., Chan Z. Hydrogen sulfide regulates abiotic stress tolerance and biotic stress resistance in Arabidopsis. J. Integr. Plant Biol. 2015;57:628–640. doi: 10.1111/jipb.12302. [DOI] [PubMed] [Google Scholar]
  • 66.Zandalinas S.I., Fichman Y., Devireddy A.R., Sengupta S., Azad R.K., Mittler R. Systemic signaling during abiotic stress combination in plants. Proc. Natl. Acad. Sci. U. S. A. 2020;117:13810–13820. doi: 10.1073/pnas.200507711. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 67.Lippert A.R., New E.J., Chang C.J. Reaction-based fluorescent probes for selective imaging of hydrogen sulfide in living cells. J. Am. Chem. Soc. 2011;26:10078–10080. doi: 10.1021/ja203661j. [DOI] [PubMed] [Google Scholar]
  • 68.Sasakura K., Hanaoka K., Shibuya N., Mikami Y., Kimura Y., Komatsu T., Ueno T., Terai T., Kimura H., Nagano T. Development of a highly selective fluorescence probe for hydrogen sulfide. J. Am. Chem. Soc. 2011;133:18003–18005. doi: 10.1021/ja207851s. [DOI] [PubMed] [Google Scholar]
  • 69.Lin V.S., Lippert A.R., Chang C.J. Cell-trappable fluorescent probes for endogenous hydrogen sulfide signaling and imaging H2O2-dependent H2S production. Proc Natl Acad Sci U.S.A. 2013;18:7131–7135. doi: 10.1073/pnas.1302193110. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 70.Peng B., Chen W., Liu C., Rosser E.W., Pacheco A., Zhao Y., Aguilar H.C., Xian M. Fluorescent probes based on nucleophilic substitution-cyclization for hydrogen sulfide detection and bioimaging. Chemistry. 2014;20:1010–1016. doi: 10.1002/chem.201303757. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 71.Wang Jian, Xie H., Li H., Wang R., Zhang B., Ren T., Hua J., Chen N. NIR fluorescent probe for in situ bioimaging of endogenous H2S in rice roots under Al3+ and flooding stresses. J. Agric. Food Chem. 2021;69:14330–14339. doi: 10.1021/acs.jafc.1c05247. [DOI] [PubMed] [Google Scholar]
  • 72.Liang K., Li Y., Zeng F., Wu S. A near-infrared fluorescent probe for detection of H2S and its application in monitoring meat freshness and plant growth under aluminium-induced stress. New J. Chem. 2023;47:2643–2650. doi: 10.1039/D2NJ05750F. [DOI] [Google Scholar]
  • 73.Ortega J.A., Ortega J.M., Julian D. Hypotaurine and sulfhydryl-containing antioxidants reduce H2S toxicity in erythrocytes from a marine invertebrate. J. Exp. Biol. 2008;211:3816–3825. doi: 10.1242/jeb.021303. [DOI] [PubMed] [Google Scholar]
  • 74.Li L., Whiteman M., Guan Y.Y., Neo K.L., Cheng Y., Lee S.W., Zhao Y., Baskar R., Tan C.H., Moore P.K. Characterization of a novel, water-soluble hydrogen sulfide-releasing molecule (GYY4137): new insights into the biology of hydrogen sulfide. Circulation. 2008;117:2351–2360. doi: 10.1161/CIRCULATIONAHA.107.753467. [DOI] [PubMed] [Google Scholar]
  • 75.Ogunniyi A.D., Kopecki Z., Hickey E.E., Khazandi M., Peel E., Belov K., Boileau A., Garg S., Venter H., Chan W.Y., Hill P.B., Page S.W., Cowin A.J., Trott D.J. Bioluminescent murine models of bacterial sepsis and scald wound infections for antimicrobial efficacy testing. PLoS One. 2018;13 doi: 10.1371/journal.pone.0200195. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 76.Gahlowt P., Tripathi D.K., Corpas F.J., Gupta R., Singh V.P. Nitric oxide working: no worries about heat stress. Trends Plant Sci. 2023;28:867–869. doi: 10.1016/j.tplants.2023.05.006. [DOI] [PubMed] [Google Scholar]
  • 77.Li Z.-G., Fang J.-R., Bai S.-J. Hydrogen sulfide signaling in plant response to temperature stress. Front. Plant Sci. 2024;15 doi: 10.3389/fpls.2024.1337250. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 78.Scuffi D., Pantaleno R., Schiel P., Niemeier J.-O., Costa A., Schwarzländer M., Laxalt A.M., García-Mata C. Hydrogen sulfide modulates flagellin-induced stomatal immunity. Plant Physiol. 2025;199 doi: 10.1093/plphys/kiaf561. kiaf561. [DOI] [PubMed] [Google Scholar]
  • 79.Maidment J.H.R., Moseler A. Small molecule, big impact: H2S contributes to pathogen-induced stomatal closure. Plant Physiol. 2025;199 doi: 10.1093/plphys/kiaf530. kiaf530. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 80.Zipfel C., Robatzek S., Navarro L., Oakeley E.J., Jones J.D.G., Felix G., Boller T. Bacterial disease resistance in Arabidopsis through flagellin perception. Nature. 2004;428:764–767. doi: 10.1038/nature02485. [DOI] [PubMed] [Google Scholar]
  • 81.Li Z.G., Yang S.Z., Long W.B., Yang G.X., Shen Z.Z. Hydrogen sulphide may be a novel downstream signal molecule in nitric oxide-induced heat tolerance of maize (Zea mays L.) seedlings. Plant Cell Environ. 2013;36:1564–1572. doi: 10.1111/pce.12092. [DOI] [PubMed] [Google Scholar]
  • 82.Zandalinas S.I., Fichman Y., Mittler R. Vascular bundles mediate systemic reactive oxygen signaling during light stress. Plant Cell. 2020;32:3425–3435. doi: 10.1105/tpc.20.00453. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 83.Pnueli L., Liang H., Rozenberg M., Mittler R. Growth suppression, altered stomatal responses, and augmented induction of heat shock proteins in cytosolic ascorbate peroxidase (Apx1)-deficient Arabidopsis plants. Plant J. 2003;34:187–203. doi: 10.1046/j.1365-313X.2003.01715.x. [DOI] [PubMed] [Google Scholar]
  • 84.Zandalinas S.I., Mittler R. Vascular and nonvascular transmission of systemic reactive oxygen signals during wounding and heat stress. Plant Physiol. 2021;186:1721–1733. doi: 10.1093/plphys/kiab157. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 85.Mukherjee S., Corpas F.J. Crosstalk among hydrogen sulfide (H2S), nitric oxide (NO) and carbon monoxide (CO) in root-system development and its rhizosphere interactions: a gaseous interactome. Plant Physiol. Biochem. 2020;155:800–814. doi: 10.1016/j.plaphy.2020.08.020. [DOI] [PubMed] [Google Scholar]
  • 86.Mishra V., Singh P., Tripathi D.K., Corpas F.J., Singh V.P. Nitric oxide and hydrogen sulfide: an indispensable combination for plant functioning. Trends Plant Sci. 2021;26:1270–1285. doi: 10.1016/j.tplants.2021.07.016. [DOI] [PubMed] [Google Scholar]
  • 87.Lindermayr C., Durner J. S-nitrosylation in plants: pattern and function. J. Proteonomics. 2009;73:1–9. doi: 10.1016/j.jprot.2009.07.002. [DOI] [PubMed] [Google Scholar]
  • 88.Zhou M., Zhang J., Shen J., Zhou H., Zhao D., Gotor C., Romero L.C., Fu L., Li Z., Yang J., Shen W., Yuan X., Xie Y. Hydrogen sulfide-linked persulfidation of ABI4 controls ABA responses through the transactivation of MAPKKK18 in Arabidopsis. Mol. Plant. 2021;14:921–936. doi: 10.1016/j.molp.2021.03.007. [DOI] [PubMed] [Google Scholar]
  • 89.Foyer C.H., Hanke G. ROS production and signalling in chloroplasts: cornerstones and evolving concepts. Plant J. 2022;111:642–661. doi: 10.1111/tpj.15856. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 90.Miller G., Suzuki N., Rizhsky L., Hegie A., Koussevitzky S., Mittler R. Double mutants deficient in cytosolic and thylakoid ascorbate peroxidase reveal a complex mode of interaction between reactive oxygen species, plant development, and response to abiotic stresses. Plant Physiol. 2007;144:1777–1785. doi: 10.1104/pp.107.101436. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 91.Correa-Aragunde N., Foresi N., Delledonne M., Lamattina L. Auxin induces redox regulation of ascorbate peroxidase 1 activity by S-nitrosylation/denitrosylation balance resulting in changes of root growth pattern in Arabidopsis. J. Exp. Bot. 2013;64:3339–3349. doi: 10.1093/jxb/ert172. [DOI] [PubMed] [Google Scholar]
  • 92.Niemeier J.O., Barreto P., Morgan B., Schwarzländer M. In vivo monitoring of energy metabolism with genetically encoded fluorescent biosensors. Annu. Rev. Plant Biol. 2026;77:16–27. doi: 10.1146/annurev-arplant-071425-085632. Epub ahead of print. PMID: 41785293. [DOI] [PubMed] [Google Scholar]

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