Next Article in Journal
Transcriptome and Physiological Analyses of a Navel Orange Mutant with Improved Drought Tolerance and Water Use Efficiency Caused by Increases of Cuticular Wax Accumulation and ROS Scavenging Capacity
Next Article in Special Issue
Crosstalk between Melatonin and Reactive Oxygen Species in Plant Abiotic Stress Responses: An Update
Previous Article in Journal
Photopharmacology of Antimitotic Agents
Previous Article in Special Issue
Hydrogen Sulfide Alleviates Manganese Stress in Arabidopsis
 
 
Search for Articles:
Title / Keyword
Author / Affiliation / Email
Journal
Article Type
 
 
Advanced Search
 
Section
Special Issue
Volume
Issue
Number
Page
 
Journals
IJMS
Volume 23
Issue 10
10.3390/ijms23105648
ijms-logo
Submit to this Journal Review for this Journal Propose a Special Issue
► Article Menu

Article Menu

share Share announcement Help format_quote Cite question_answer Discuss in SciProfiles thumb_up
...
Endorse textsms
...
Comment
first_page
settings
Order Article Reprints
Font Type:
Arial Georgia Verdana
Font Size:
Aa Aa Aa
Line Spacing:
Column Width:
Background:
Article

H2S in Horticultural Plants: Endogenous Detection by an Electrochemical Sensor, Emission by a Gas Detector, and Its Correlation with L-Cysteine Desulfhydrase (LCD) Activity

by
María A. Muñoz-Vargas
,
Salvador González-Gordo
,
José M. Palma
and
Francisco J. Corpas
*
Group of Antioxidants, Free Radicals and Nitric Oxide in Biotechnology, Food and Agriculture, Department of Biochemistry, Cell and Molecular Biology of Plants, Estación Experimental del Zaidín, Spanish National Research Council (CSIC), C/Profesor Albareda 1, E-18008 Granada, Spain
*
Author to whom correspondence should be addressed.
Int. J. Mol. Sci. 2022, 23(10), 5648; https://doi.org/10.3390/ijms23105648
Submission received: 27 April 2022 / Revised: 13 May 2022 / Accepted: 17 May 2022 / Published: 18 May 2022
(This article belongs to the Special Issue Hydrogen Sulfide and Reactive Oxygen Species, Antioxidant Defense, Abiotic Stress Tolerance Mechanisms in Plants)
Browse Figures
Versions Notes

Abstract

:
H2S has acquired great attention in plant research because it has signaling functions under physiological and stress conditions. However, the direct detection of endogenous H2S and its potential emission is still a challenge in higher plants. In order to achieve a comparative analysis of the content of H2S among different plants with agronomical and nutritional interest including pepper fruits, broccoli, ginger, and different members of the genus Allium such as garlic, leek, Welsh and purple onion, the endogenous H2S and its emission was determined using an ion-selective microelectrode and a specific gas detector, respectively. The data show that endogenous H2S content range from pmol to μmol H2S · g−1 fresh weight whereas the H2S emission of fresh-cut vegetables was only detected in the different species of the genus Allium with a maximum of 9 ppm in garlic cloves. Additionally, the activity and isozymes of the L-cysteine desulfhydrase (LCD) were analyzed, which is one of the main enzymatic sources of H2S, where the different species of the genus Allium showed the highest activities. Using non-denaturing gel electrophoresis, the data indicated the presence of up to nine different LCD isozymes from one in ginger to four in onion, leek, and broccoli. In summary, the data indicate a correlation between higher LCD activity with the endogenous H2S content and its emission in the analyzed horticultural species. Furthermore, the high content of endogenous H2S in the Allium species supports the recognized benefits for human health, which are associated with its consumption.

1. Introduction

H2S is a key signaling molecule that plays multiple functions in many physiological and pathological processes in humans, regulating the basal metabolism, central nervous system, blood pressure, gastrointestinal motility, inflammation, the immune system, or cancer, among others [1,2,3,4,5,6,7]. In higher plants, H2S also has relevant functions due to its direct or indirect implication in physiological functions including seed germination, root development, plant growth, stomatal closure, senescence, and fruit ripening as well as in the mechanism of response against adverse environmental conditions [8,9,10,11,12,13]. H2S is part of the sulfur metabolism being enzymatically generated by different enzymes present in diverse subcellular compartments including cytosol, plastids, and mitochondria [14]. At the biochemical level, H2S mediates the regulation of protein function by a posttranslational modification designated persufidation which involved the thiol group of cysteine residues from target proteins [15,16,17]. However, the detection of endogenous hydrogen sulfide and its possible emission continues to be a scientific challenge due to the complex biochemistry that is affected by its interaction with peptides and proteins, pH, cellular location, type of biological samples, etc. Different methodologies allow the detection of endogenous H2S in different types of biological samples such as high-performance liquid chromatography (HPLC), gas chromatography (GC), colorimetric, specific fluorescence probes, ion-selective electrode (ISE), amperometric (polarographic) H2S sensor, ozone-based chemiluminescence detection among others and all have advantages and disadvantages as well as different detection limits [18,19,20,21].
L-Cysteine desulfhydrase (LCD, EC 4.4.1.28) is considered one of the main cytosolic enzymatic sources of H2S in Arabidopsis cells which is also generated by other enzymes such as the chloroplastic sulfite reductase (SiR, EC 1.8.7.1) or the mitochondrial bifunctional D-cysteine desulfhydrase/1-aminocyclopropane-1-carboxylate deaminase (DCDES1, EC 4.4.1.15) and D-cysteine desulfhydrase 2 (DCDES2, EC 4.4.1.15) [14,22,23]. The LCD catalyzes the following reaction: L-cysteine + H2O → pyruvate + NH4+ + H2S + H+, and it requires pyridoxal 5′-phosphate (PLP) as cofactor [23,24]. This enzyme is involved in diverse processes such as root development [25], stomatal closure [26,27,28], leaf senescence [29], fruit ripening [30,31], and response to diverse stresses [32].
Aiming to correlate the potential relationship between endogenous H2S, its potential emission, and the activity of the LCD, the present study provides a comparative analysis of these parameters in some horticultural plants such as pepper fruits, broccoli, ginger, fennel, eggplant, leek, garlic, Welsh and onion, which are relevant to human nutrition. The Allium species, particularly garlic cloves, are horticultural plants that present the highest values of H2S that is well correlated with its very active metabolism of organosulfur compounds such as allicin, allyl sulfides, allyl thiosulfinate, ajoene, and S-allyl cysteine among others.

2. Results

Figure 1a–c illustrates the endogenous H2S content in different horticultural plant species which was measured using an ion-selective microelectrode (Arrow H2S™) H2S measurement system. Accordingly, three well-differentiated data groups can be distinguished, while the species of the Allium genus give H2S values in the range of μmol H2S · g−1 FW, the leek with 1 μmol H2S · g−1 FW being the species with the highest content, followed by garlic cloves with 0.54 μmol H2S · g−1 FW. Other groups are in the range of nmol H2S · g−1 FW such as broccoli but also in the range of pmol H2S · g−1 FW such as pepper fruit at different stages of ripening green and red. On the other hand, in fennel and eggplant samples the H2S detected was even lower, so they were not used for subsequent studies.
As part of the characterization of H2S in these horticultural plants, the detection of H2S gas emission was performed using 300 g of cut materials. Figure 2 indicates that the species of the genus Allium were the only ones that allowed it to be detected. It is noteworthy that the emission of H2S is relatively quite fast and is maintained over time. Thus, leek reaches a maximum peak of 3.3 ppm of H2S after 30 min, followed by spring onion with a maximum peak of 7.4 ppm after 75 min, purple onion with 8.1 ppm after 90 min, and finally garlic with 9.0 after 180 min. It is noteworthy that the garlic maintained the emission of H2S up to 0.9 ppm after 21 h.
Assuming that the L-cysteine desulfhydrase (LCD) activity is considered the enzyme that most contributes to the generation of H2S in the cell, its activity was measured spectrophotometrically and also in polyacrylamide gels under non-denaturing conditions. Figure 3a shows that species of the Allium genus have higher LCD activity, with garlic having the highest one, followed by broccoli, pepper, and ginger. On the other hand, Figure 3b shows the LCD isozymes profile in the different analyzed plant species. They were designated as I to IX according to their increasing electrophoretic mobility in the non-denaturing polyacrylamide gel. The number and relative abundance were quite different, while a single LCD isozyme is identified in ginger, three isozymes appear in garlic, broccoli, and green peppers, and up to four LCD isozymes in purple and Welsh onions. In pepper fruits, it is remarkable that the number of isozymes changes with ripening, having three in green pepper fruits and only one in red peppers.

3. Discussion

H2S is recognized as a key molecule with a signaling function in animal and plant cells, with similar regulatory properties to those exerted by nitric oxide (NO) in higher plants under physiological and stress conditions [9,33,34,35,36,37,38]. With the aim of obtaining a better understanding of this molecule in different plants with agronomic interest, the endogenous content, its emission, as well as its possible correlation with LCD activity have been comparatively studied, since it is considered the most relevant enzyme in the H2S production in plant cells.
One of the difficulties in determining the endogenous H2S content in a specific sample is based on its chemistry because, being a weak acid, it can be dissociated to hydrosulfide (HS–) and sulfide (S2−) anions in an aqueous solution according to the following equations: H2S(gas) ↔ H2S(aqueous solution) ↔ HS− + H+ ↔ S2− + 2H+. Other considerations that should be taken into account are the pH of the medium, the nature of biological samples, that H2S can interact with thiol groups present in peptides and proteins, the selectivity of the technical approach as well as the experimental conditions [2,15,18,39]. Otherwise, the development of new approaches such as the specific fluorescent probes has allowed the detection of H2S at the subcellular level by bioimaging approaches [40,41,42,43]; however, its quantification is still a challenge. In an earlier study in cucumber (Cucumis sativus L.) plants, an H2S concentration of 8 nmol · min−1 g−1 FW that was light-dependent [44] was reported and, after fumigation with SO2, the H2S content was 0.02 to 0.2 ng · g−1 dry weight [45]. In Arabidopsis thaliana and faba bean (Vicia faba) leaves, using a micro sulfide ion electrode, an H2S content between 1 to 5 μmol · L−1 was reported [46,47]. More recently, Jin et al. [48], using both methylene blue and electrode methods, determined the H2S content of 17 species in different developmental stages and different organs and they showed a wider range, from 0.177 to 0.708 μmol · g−1 FW, in which Platycladus sp. in the cypress family had the highest content whereas tobacco had the lowest content. Consequently, our data in the assayed horticultural plants are in good agreement with all these previous reports. Although the values of H2S content determined with the micro sulfide ion electrode provide relative measurements accurately and reliably, the difficulty to measure the H2S content when these values are low to 0.1 μmol · g−1 FW should be mentioned and, consequently, the obtained values should be interpreted with caution.
The emission of volatile H2S is another aspect that could have great relevance in higher plants, although the available information is still very scarce. In an earlier study, Sekiya et al. [49], analyzed the H2S emission by gas chromatography in leaf discs incubated in the presence of 10 mM L-Cys from nine species (Cucumis sativus, Cucurbita pepo, Nicotiana tabacum, Coleus blumei, Beta vulgaris, Phaseolus vulgaris, Medicago sativa, Hordeum vulgare, and Gossypium hirsutum) and they found an emission around 40 pmol H2S · min−1 cm−2; however, the H2S emission was not observed in the presence of D-Cys. Rennenberg et al. [50], using a flame photometric sulfur analyzer, reported an H2S emission between 38 to 91 pmol H2S · min−1 cm−2 pumpkin leaf area. In our experimented conditions, we only observed H2S emission in the species of the genus Allium, and this emission was maintained in the time, particularly in garlic which was up to 9 ppm. It is well known that these species of the genus Allium have a characteristic smell and taste which is due to the sulfur-containing volatile flavor compounds. These volatile constituents are generated by the action of the enzyme alliinase, with a molecular mass between 13 and 35 kDa depending on the Allium species, when plant tissue is disrupted, the alliinase catalyzes the conversion of odorless S-alk(en)yl-L-cysteine sulfoxides (SACs), known as alliin, into volatile smelling thiosulfinates [51,52]. These SACs are synthesized from glutathione in the cytosol and when the compartmentalization of the alliinase, present in the vacuole, is broken down, it allows the metabolization of these SACs and its emission in volatile thiosulfinates [53,54,55]. In fact, these groups of sulfur-containing natural products present in the different species of Allium are correlated with the benefits associated with human health when they are part of our diet [52,56]. Thus, it is well documented that garlic consumption reduces some risk factors related to cardiovascular diseases such as high blood pressure, high cholesterol, platelet aggregation, blood coagulation, and the increased content of reactive oxygen species (ROS) [52,57,58,59,60]. The detected H2S emission in the different species of Allium is also well correlated with the beneficial effects exerted on human health [59]. In plants, these sulfur-containing molecules including H2S have great relevance in the resistance of crops against diverse fungal diseases [61,62].
L-cysteine desulfhydrase (LCD) catalyzes the desulfuration of L-Cys to generate H2S and is considered one of the main sources of this molecule in the cytosol of plant cells [63]. As was mentioned, biochemical and molecular approaches have revealed that LCD is implicated in diverse processes such as seed germination [64], root development [25,65], stomatal closure [26,27,66], drought tolerance [67], and fruit ripening [31,68].
In our experimental conditions, it is well correlated with the total LCD activity found in the Allium species with the highest level of endogenous H2S content as well as its emission. However, the available information about the number of LCD isozymes present in a specific tissue or plant species and its corresponding functions is to our knowledge very scarcer. Considering all analyzed species and tissues, the present data indicate the existence of nine LCD isozymes indicating the great diversity in the number and relative abundance which seems to support the differential potential physiological regulatory functions that they could have. For example, in the pepper fruit samples, it was observed that total LCD activity was downregulated during ripening from green to red fruits, which was well correlated with the diminishment of the LCD isozymes since two of them were not detected in red fruits.

4. Materials and Methods

4.1. Plant Material

California-type sweet pepper (Capsicum annuum L., cv. Melchor) fruits were collected from plastic-covered experimental greenhouses (Syngenta Seeds, Ltd., El Ejido, Almería, Spain) whereas the other plants were acquired in the local market including broccoli (Brassica oleracea var. Itálica), ginger (Zingiber officinale) rhizome, fennel (Foeniculum vulgare), eggplant (Solanum melongena), leek (Allium ampeloprasum var. porrum), Welsh onion (Allium fistulosum), purple onion (Allium cepa) and garlic (Allium sativum L.) cloves.

4.2. Preparation of Plant Extracts

Plant samples were ground to a fine powder in liquid N2 using an IKA® A11 basic analytical mill. The resulting powder was suspended in 0.1 M Tris-HCl buffer, pH 7.5, containing 1 mM EDTA, 0.1% (v/v) Triton X-100, 10% (v/v) glycerol to a final plant material/buffer (w/v) ratio of 1:1 for pepper, fennel, and leek fruit; 1:2 for eggplant, garlic, ginger, and broccoli; and, 2:1 for red onion and Welsh onion. Homogenates were then filtered through two layers of Miracloth and centrifuged at 27,000× g for 20 min. The supernatants were used for subsequent analyses. For H2S endogenous quantification, the supernatants were mixed with antioxidant buffer (2 M NaOH, 170 mM sodium ascorbate, and 180 mM EDTA). In the case of enzymatic activity, the extraction buffer was similar but with a pH of 8.0. Protein content was determined by a standard Bradford assay using a reagent (Bio-Rad Laboratories, Hercules, CA, USA).

4.3. Endogenous H2S Quantification

H2S was measured in plant extracts for 5 min at 25 °C using a micro sulfide ion electrode (LIS-146AGSCM; Lazar Research Laboratories) attached to a voltage meter (Lazar Research Lab. Inc., Los Angeles, CA, USA, model ISM-146 H2S-XS). H2S concentrations were calculated from a standard curve made with sodium sulfide (Na2S) according to the micro-electrode manufacturer’s instructions.

4.4. H2S Gas Emission Hydrogen Sulfide Gas Detector

H2S gas emission was recorded using a high accuracy H2S sensor gas analyzer portable device (NOBGP brand, model TK3Bcb4T78 model), which can measure H2S gas in a range between 0~100 μmol · mL−1, a resolution of 0.1 with an accuracy less than or equal to ±5% full scale. In all cases, 300 g of fresh plant samples were cut into homogeneous pieces and placed in a methacrylate hermetic box (10 mm-thickness walls): 25 (large) × 25 (width) × 30 (height) cm = 15.34 L, furnished with a lid made on the same material. The H2S gas detector was placed into the box and the H2S emission was recorded for 18 h. In the case of garlic, the cloves were used but for leek and Welsh onion, the stringy roots and dark green leaves were chopped off.

4.5. Spectrophotometric Assay and In-Gel Isozyme Profile of L-Cysteine Desulfrydrase (LCD) Activity

L-cysteine desulfhydrase (LCD, E.C. 4.4.1.28) activity was spectrophotometrically determined by the release of H2S from L-Cys as described previously [69,70]. Briefly, the enzyme assay contained 1 mM L-Cys, 100 mM Tris-HCl, pH 8.0, 1 mM dithiothreitol, and plant extract in a final volume of 1 mL. After 15 min incubation at 37 °C, the reaction was stopped by the addition of 100 μL of 30 mM FeCl3 prepared in 1.2 N HCl and 100 μL of 20 mM N,N-dimethyl-p-phenylenediamine dihydrochloride prepared in 7.2 N HCl. The formation of methylene blue was measured at 670 nm, and the enzyme activity was calculated using the extinction coefficient of 15 × 106 cm2 mol−1.
For in-gel isozyme profile analysis, protein samples were separated using non-denaturing polyacrylamide gel electrophoresis (PAGE) on 8% acrylamide gels. After the electrophoresis, the gels were incubated in the dark in a staining buffer containing Tris-HCl 100 mM, pH 7.5, L-cysteine 20 mM, lead acetate 0.4 mM, pyridoxal 5′-phosphate hydrate 50 µM and β-mercaptoethanol 20 mM until the appearance of brown bands [24,71].

5. Conclusions

H2S is a signaling molecule in both animal and plant cells. The analysis of its endogenous content as to its possible emission in higher plants to determine its physiological functions and in response to environmental stresses has been a challenge for years [72]. However, the information concerning H2S in horticultural species is still scarce; therefore, the present study provides new information on the content and emission of H2S in horticultural plants, particularly in the species of the Allium genus. This may be of great importance in horticultural crops, considering that H2S applied exogenously has been shown to exert multiple benefits for vegetables and fruits, since it has the capacity to preserve their quality during postharvest storage and prevents infections by pathogens because H2S stimulates phytohormone, reactive oxygen, and nitrogen metabolism [73,74,75,76,77,78]. Furthermore, the identification of different LCD isozymes in the analyzed horticultural species indicates the relevance of this enzyme in H2S metabolism and raises new questions about the specific function of each isozyme which could be modulated by environmental conditions or physiological processes such as was observed in the ripening of pepper fruits. Allium species have been recognized for a long time to have healthy properties [79], and among the sulfur compounds that it contains, H2S seems to be of great relevance [58,80]. On the other hand, considering the high H2S emission of Allium species, they should be regarded as a potential source of this gas for its possible biotechnological application in the horticulture industry since it can extend the quality of vegetables and fruits during postharvest storage.

Author Contributions

M.A.M.-V. and S.G.-G. performed biochemical analyses. F.J.C. and J.M.P. designed the work, drove and coordinated the tasks. F.J.C. wrote the first draft of the manuscript. All authors have read and agreed to the published version of the manuscript.

Funding

Our research is supported by a European Regional Development Fund cofinanced grants from the Spanish Ministry of Science and Innovation (PID2019-103924GB-I00) and Junta de Andalucía (P18-FR-1359), Spain.

Institutional Review Board Statement

Not applicable.

Informed Consent Statement

Not applicable.

Acknowledgments

M.A.M.-V. and S.G.-G. acknowledge a ‘Formación de Personal Investigador’ contract (PRE2020-093882 and BES-2016-078368, respectively) from the Ministry of Science and Innovation, Spain. The valuable technical assistance of Carmelo Ruiz-Torres is deeply acknowledged.

Conflicts of Interest

The authors declare no conflict of interest.

References

  1. Banerjee, R.; Vitvitsky, V.; Garg, S.K. The undertow of sulfur metabolism on glutamatergic neurotransmission. Trends Biochem. Sci. 2008, 33, 413–419. [Google Scholar] [CrossRef] [PubMed]
  2. Kabil, O.; Vitvitsky, V.; Banerjee, R. Sulfur as a signaling nutrient through hydrogen sulfide. Annu Rev. Nutr. 2014, 34, 171–205. [Google Scholar] [CrossRef] [Green Version]
  3. Szabo, C. Hydrogen sulfide, an endogenous stimulator of mitochondrial function in cancer cells. Cells 2021, 10, 220. [Google Scholar] [CrossRef] [PubMed]
  4. Parfenova, H.; Liu, J.; Hoover, D.T.; Fedinec, A.L. Vasodilator effects of sulforaphane in cerebral circulation: A critical role of endogenously produced hydrogen sulfide and arteriolar smooth muscle KATP and BK channels in the brain. J. Cereb. Blood Flow Metab. 2020, 40, 1987–1996. [Google Scholar] [CrossRef] [PubMed]
  5. Kumar, R.; Banerjee, R. Regulation of the redox metabolome and thiol proteome by hydrogen sulfide. Crit. Rev. Biochem. Mol. Biol. 2021, 56, 221–235. [Google Scholar] [CrossRef]
  6. Kimura, H. Hydrogen sulfide (H2S) and polysulfide (H2Sn) signaling: The First 25 Years. Biomolecules 2021, 11, 896. [Google Scholar] [CrossRef]
  7. Deng, G.; Muqadas, M.; Adlat, S.; Zheng, H.; Li, G.; Zhu, P.; Nasser, M.I. Protective effect of hydrogen sulfide on cerebral ischemia-reperfusion injury. Cell. Mol. Neurobiol. 2022, in press. [CrossRef]
  8. 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. [Google Scholar] [CrossRef]
  9. 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. [Google Scholar] [CrossRef]
  10. Rai, P.; Singh, V.P.; Peralta-Videa, J.; Tripathi, D.K.; Sharma, S.; Corpas, F.J. Hydrogen sulfide (H2S) underpins the beneficial silicon effects against the copper oxide nanoparticles (CuO NPs) phytotoxicity in Oryza sativa seedlings. J. Hazard. Mater. 2021, 415, 124907. [Google Scholar] [CrossRef]
  11. Raza, A.; Tabassum, J.; Mubarik, M.S.; Anwar, S.; Zahra, N.; Sharif, Y.; Hafeez, M.B.; Zhang, C.; Corpas, F.J.; Chen, H. Hydrogen sulfide: An emerging component against abiotic stress in plants. Plant Biol. 2021, in press. [CrossRef] [PubMed]
  12. Zhang, J.; Zhou, M.; Zhou, H.; Zhao, D.; Gotor, C.; Romero, L.C.; Shen, J.; Ge, Z.; Zhang, Z.; Shen, W.; et al. Hydrogen sulfide, a signaling molecule in plant stress responses. J. Integr. Plant Biol. 2021, 63, 146–160. [Google Scholar] [CrossRef] [PubMed]
  13. Aroca, A.; Zhang, J.; Xie, Y.; Romero, L.C.; Gotor, C. Hydrogen sulfide signaling in plant adaptations to adverse conditions: Molecular mechanisms. J. Exp. Bot. 2021, 72, 5893–5904. [Google Scholar] [CrossRef] [PubMed]
  14. González-Gordo, S.; Palma, J.M.; Corpas, F.J. Appraisal of H2S metabolism in Arabidopsis thaliana: In silico analysis at the subcellular level. Plant Physiol. Biochem. 2020, 155, 579–588. [Google Scholar] [CrossRef]
  15. Filipovic, M.R.; Zivanovic, J.; Alvarez, B.; Banerjee, R. Chemical biology of H2S signaling through persulfidation. Chem. Rev. 2018, 118, 1253–1337. [Google Scholar] [CrossRef]
  16. Corpas, F.J.; González-Gordo, S.; Muñoz-Vargas, M.A.; Rodríguez-Ruiz, M.; Palma, J.M. The modus operandi of hydrogen sulfide(H2S)-dependent protein persulfidation in higher plants. Antioxidants 2021, 10, 1686. [Google Scholar] [CrossRef]
  17. Corpas, F.J.; González-Gordo, S.; Palma, J.M. Nitric oxide and hydrogen sulfide modulate the NADPH-generating enzymatic system in higher plants. J. Exp. Bot. 2021, 72, 830–847. [Google Scholar] [CrossRef]
  18. Zhao, D.; Zhang, J.; Zhou, M.; Zhou, H.; Gotor, C.; Romero, L.C.; Shen, J.; Yuan, X.; Xie, Y. Current approaches for detection of hydrogen sulfide and persulfidation in biological systems. Plant Physiol. Biochem. 2020, 155, 367–373. [Google Scholar] [CrossRef]
  19. Luo, Y.; Zuo, Y.; Shi, G.; Xiang, H.; Gu, H. Progress on the reaction-based methods for detection of endogenous hydrogen sulfide. Anal. Bioanal. Chem. 2021, 414, 2809–2839. [Google Scholar] [CrossRef]
  20. Quan, F.S.; Lee, G.J. Analytical methods for detection of gasotransmitter hydrogen sulfide released from live cells. BioMed Res. Int. 2021, 2021, 5473965. [Google Scholar] [CrossRef]
  21. Li, B.; Kim, Y.L.; Lippert, A.R. Chemiluminescence measurement of reactive sulfur and nitrogen species. Antioxid. Redox Signal. 2022, 36, 337–353. [Google Scholar] [CrossRef] [PubMed]
  22. Feldman-Salit, A.; Veith, N.; Wirtz, M.; Hell, R.; Kummer, U. Distribution of control in the sulfur assimilation in Arabidopsis thaliana depends on environmental conditions. New Phytol. 2019, 222, 1392–1404. [Google Scholar] [CrossRef] [PubMed]
  23. Kurmanbayeva, A.; Bekturova, A.; Soltabayeva, A.; Oshanova, D.; Nurbekova, Z.; Srivastava, S.; Tiwari, P.; Dubey, A.K.; Sagi, M. Active O-acetylserine-(thiol) lyase A and B confer improved selenium resistance and degrade l-Cys and l-SeCys in Arabidopsis. J. Exp. Bot. 2022, 73, 2525–2539. [Google Scholar] [CrossRef]
  24. Muñoz-Vargas, M.A.; Rodríguez-Ruiz, M.; González-Gordo, S.; Palma, J.M.; Corpas, F.J. Analysis of Plant L-Cysteine desulfhydrase (LCD) isozymes by non-denaturing polyacrylamide gel electrophoresis. In Plant Abiotic Stress Signaling. Methods in Molecular Biology; Couée, I., Ed.; Humana: New York, NY, USA, 2023; in press. [Google Scholar]
  25. Mei, Y.; Zhao, Y.; Jin, X.; Wang, R.; Xu, N.; Hu, J.; Huang, L.; Guan, R.; Shen, W. L-Cysteine desulfhydrase-dependent hydrogen sulfide is required for methane-induced lateral root formation. Plant Mol. Biol. 2019, 99, 283–298. [Google Scholar] [CrossRef]
  26. Scuffi, D.; Álvarez, C.; Laspina, N.; Gotor, C.; Lamattina, L.; García-Mata, C. Hydrogen sulfide generated by L-cysteine desulfhydrase acts upstream of nitric oxide to modulate abscisic acid-dependent stomatal closure. Plant Physiol. 2014, 166, 2065–2076. [Google Scholar] [CrossRef] [Green Version]
  27. Shen, J.; Zhang, J.; Zhou, M.; Zhou, H.; Cui, B.; Gotor, C.; Romero, L.C.; Fu, L.; Yang, J.; Foyer, C.H.; et al. Persulfidation-based modification of cysteine desulfhydrase and the NADPH oxidase RBOHD controls guard cell abscisic acid signaling. Plant Cell 2020, 32, 1000–1017. [Google Scholar] [CrossRef] [PubMed]
  28. Ma, Y.; Shao, L.; Zhang, W.; Zheng, F. Hydrogen sulfide induced by hydrogen peroxide mediates brassinosteroid-induced stomatal closure of Arabidopsis thaliana. Funct. Plant Biol. 2021, 48, 195–205. [Google Scholar] [CrossRef]
  29. Hu, K.; Peng, X.; Yao, G.; Zhou, Z.; Yang, F.; Li, W.; Zhao, Y.; Li, Y.; Han, Z.; Chen, X.; et al. Roles of a cysteine desulfhydrase LCD1 in regulating leaf senescence in tomato. Int. J. Mol. Sci. 2021, 22, 13078. [Google Scholar] [CrossRef]
  30. Muñoz-Vargas, M.A.; González-Gordo, S.; Cañas, A.; López-Jaramillo, J.; Palma, J.M.; Corpas, F.J. Endogenous hydrogen sulfide (H2S) is up-regulated during sweet pepper (Capsicum annuum L.) fruit ripening. In vitro analysis shows that NADP-dependent isocitrate dehydrogenase (ICDH) activity is inhibited by H2S and NO. Nitric Oxide 2018, 81, 36–45. [Google Scholar] [CrossRef]
  31. Hu, K.D.; Zhang, X.Y.; Yao, G.F.; Rong, Y.L.; Ding, C.; Tang, J.; Yang, F.; Huang, Z.Q.; Xu, Z.M.; Chen, X.Y.; et al. A nuclear-localized cysteine desulfhydrase plays a role in fruit ripening in tomato. Hortic. Res. 2020, 7, 211. [Google Scholar] [CrossRef]
  32. Kharbech, O.; Sakouhi, L.; Mahjoubi, Y.; Ben Massoud, M.; Debez, A.; Zribi, O.T.; Djebali, W.; Chaoui, A.; Mur, L.A.J. Nitric oxide donor, sodium nitroprusside modulates hydrogen sulfide metabolism and cysteine homeostasis to aid the alleviation of chromium toxicity in maize seedlings (Zea mays L.). J. Hazard. Mater. 2022, 424 Pt A, 127302. [Google Scholar] [CrossRef]
  33. Gheibi, S.; Jeddi, S.; Kashfi, K.; Ghasemi, A. Regulation of vascular tone homeostasis by NO and H2S: Implications in hypertension. Biochem. Pharmacol. 2018, 149, 42–59. [Google Scholar] [CrossRef] [PubMed]
  34. Lv, B.; Chen, S.; Tang, C.; Jin, H.; Du, J.; Huang, Y. Hydrogen sulfide and vascular regulation—An update. J. Adv. Res. 2020, 27, 85–97. [Google Scholar] [CrossRef] [PubMed]
  35. Kimura, H. Hydrogen polysulfide (H2Sn) signaling along with hydrogen sulfide (H2S) and nitric oxide (NO). J. Neural Transm. (Vienna) 2016, 123, 1235–1245. [Google Scholar] [CrossRef] [PubMed]
  36. Kimura, H. Hydrogen sulfide signalling in the CNS—Comparison with NO. Br. J. Pharmacol. 2020, 177, 5031–5045. [Google Scholar] [CrossRef] [PubMed]
  37. Corpas, F.J. Hydrogen sulfide: A new warrior against abiotic Stress. Trends Plant Sci. 2019, 24, 983–988. [Google Scholar] [CrossRef] [PubMed]
  38. Kharbech, O.; Sakouhi, L.; Ben Massoud, M.; Jose Mur, L.A.; Corpas, F.J.; Djebali, W.; Chaoui, A. Nitric oxide and hydrogen sulfide protect plasma membrane integrity and mitigate chromium-induced methylglyoxal toxicity in maize seedlings. Plant Physiol. Biochem. 2020, 157, 244–255. [Google Scholar] [CrossRef]
  39. Papenbrock, J.; Riemenschneider, A.; Kamp, A.; Schulz-Vogt, H.N.; Schmidt, A. Characterization of cysteine-degrading and H2S-releasing enzymes of higher plants—From the field to the test tube and back. Plant Biol. 2007, 9, 582–588. [Google Scholar] [CrossRef]
  40. 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. [Google Scholar] [CrossRef] [Green Version]
  41. Yu, F.; Han, X.; Chen, L. Fluorescent probes for hydrogen sulfide detection and bioimaging. Chem. Commun. 2014, 50, 12234–12249. [Google Scholar] [CrossRef]
  42. Corpas, F.J.; Barroso, J.B.; González-Gordo, S.; Muñoz-Vargas, M.A.; Palma, J.M. Hydrogen sulfide: A novel component in Arabidopsis peroxisomes which triggers catalase inhibition. J. Integr. Plant Biol. 2019, 61, 871–883. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  43. 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. [Google Scholar] [CrossRef] [PubMed]
  44. Wilson, L.G.; Bressan, R.A.; Filner, P. Light-dependent emission of hydrogen sulfide from plants. Plant Physiol. 1978, 61, 184–189. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  45. Winner, W.E.; Smith, C.L.; Koch, G.W.; Mooney, H.A.; Bewley, J.D.; Krouse, H.R. Rates of emission of H2S from plants and patterns of stable sulphur isotope fractionation. Nature 1981, 289, 672–673. [Google Scholar] [CrossRef]
  46. García-Mata, C.; Lamattina, L. Hydrogen sulphide, a novel gasotransmitter involved in guard cell signaling. New Phytol. 2010, 188, 977–984. [Google Scholar] [CrossRef]
  47. Hou, Z.H.; Liu, J.; Hou, L.X.; Li, X.; Liu, X. H2S may function downstream of H2O2 in jasmonic acid-induced stomatal closure in Vicia faba. Chin. Bull. Bot. 2011, 4, 396–406. [Google Scholar]
  48. Jin, Z.; Wang, Z.; Yang, G.; Pei, Y. Diversity of hydrogen sulfide concentration in plant: A little spark to start a prairie fire. Sci. Bull. 2018, 63, 1314–1316. [Google Scholar] [CrossRef] [Green Version]
  49. Sekiya, J.; Schmidt, A.; Wilson, L.G.; Filner, P. Emission of hydrogen sulfide by leaf tissue in response to L-cysteine. Plant Physiol. 1982, 70, 430–436. [Google Scholar] [CrossRef] [Green Version]
  50. Rennenberg, H. Role of o-acetylserine in hydrogen sulfide emission from pumpkin leaves in response to sulfate. Plant Physiol. 1983, 73, 560–565. [Google Scholar] [CrossRef] [Green Version]
  51. Krest, I.; Glodek, J.; Keusgen, M. Cysteine sulfoxides and alliinase activity of some Allium species. J. Agric. Food Chem. 2000, 48, 3753–3760. [Google Scholar] [CrossRef]
  52. Yoshimoto, N.; Saito, K. S-Alk(en)ylcysteine sulfoxides in the genus Allium: Proposed biosynthesis, chemical conversion, and bioactivities. J. Exp. Bot. 2019, 70, 4123–4137. [Google Scholar] [CrossRef] [PubMed]
  53. Lancaster, J.E.; Collin, H.A. 1981. Presence of alliinase in isolated vacuoles and of alkyl cysteine sulphoxides in the cytoplasm of bulbs of onion (Allium cepa). Plant Sci. Lett. 1981, 22, 169–176. [Google Scholar] [CrossRef]
  54. Lancaster, J.E.; Dommisse, E.M.; Shaw, M.L. Production of flavor precursors [S-alk(en)yl-l-cysteine sulphoxides] in photomixotrophic callus of garlic. Phytochemistry 1988, 27, 2123–2124. [Google Scholar] [CrossRef]
  55. Lancaster, J.E.; Reynolds, P.H.S.; Shaw, M.L.; Dommisse, E.M.; Munro, J. Intra-cellular localization of the biosynthetic pathway to flavour precursors in onion. Phytochemistry 1989, 28, 461–464. [Google Scholar] [CrossRef]
  56. Nohara, T.; Fujiwara, Y.; El-Aasr, M.; Ikeda, T.; Ono, M.; Nakano, D.; Kinjo, J. Thiolane-type sulfides from garlic, onion, and Welsh onion. J. Nat. Med. 2021, 75, 741–751. [Google Scholar] [CrossRef]
  57. Banerjee, S.K.; Maulik, S.K. Effect of garlic on cardiovascular disorders: A review. Nutr. J. 2002, 1, 4. [Google Scholar] [CrossRef]
  58. Benavides, G.A.; Squadrito, G.L.; Mills, R.W.; Patel, H.D.; Isbell, T.S.; Patel, R.P.; Darley-Usmar, V.M.; Doeller, J.E.; Kraus, D.W. Hydrogen sulfide mediates the vasoactivity of garlic. Proc. Natl. Acad. Sci. USA 2007, 104, 17977–17982. [Google Scholar] [CrossRef] [Green Version]
  59. Rose, P.; Moore, P.K.; Zhu, Y.Z. Garlic and Gaseous Mediators. Trends Pharmacol. Sci. 2018, 39, 624–634. [Google Scholar] [CrossRef]
  60. Alali, F.Q.; El-Elimat, T.; Khalid, L.; Hudaib, R.; Al-Shehabi, T.S.; Eid, A.H. Garlic for cardiovascular disease: Prevention or treatment? Curr. Pharm. Des. 2017, 23, 1028–1041. [Google Scholar] [CrossRef]
  61. Nwachukwu, I.D.; Slusarenko, A.J.; Gruhlke, M.C. Sulfur and sulfur compounds in plant defence. Nat. Prod. Commun. 2012, 7, 395–400. [Google Scholar] [CrossRef] [Green Version]
  62. Künstler, A.; Gullner, G.; Ádám, A.L.; Kolozsváriné Nagy, J.; Király, L. The versatile roles of sulfur-containing biomolecules in plant defense-a road to disease resistance. Plants 2020, 9, 1705. [Google Scholar] [CrossRef] [PubMed]
  63. Laureano-Marín, A.M.; García, I.; Romero, L.C.; Gotor, C. Assessing the transcriptional regulation of L-cysteine desulfhydrase 1 in Arabidopsis thaliana. Front. Plant Sci. 2014, 5, 683. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  64. Zhou, M.; Zhang, J.; Zhou, H.; Zhao, D.; Duan, T.; Wang, S.; Yuan, X.; Xie, Y. Hydrogen sulfide-linked persulfidation maintains protein stability of ABSCISIC ACID-INSENSITIVE 4 and delays seed germination. Int. J. Mol. Sci. 2022, 23, 1389. [Google Scholar] [CrossRef] [PubMed]
  65. Fang, T.; Cao, Z.; Li, J.; Shen, W.; Huang, L. Auxin-induced hydrogen sulfide generation is involved in lateral root formation in tomato. Plant Physiol. Biochem. 2014, 76, 44–51. [Google Scholar] [CrossRef]
  66. Zhang, J.; Zhou, M.; Ge, Z.; Shen, J.; Zhou, C.; Gotor, C.; Romero, L.C.; Duan, X.; Liu, X.; Wu, D.; et al. Abscisic acid-triggered guard cell l-cysteine desulfhydrase function and in situ hydrogen sulfide production contributes to heme oxygenase-modulated stomatal closure. Plant Cell Environ. 2020, 43, 624–636. [Google Scholar] [CrossRef]
  67. Zhou, H.; Zhou, Y.; Zhang, F.; Guan, W.; Su, Y.; Yuan, X.; Xie, Y. Persulfidation of nitrate reductase 2 is involved in l-cysteine desulfhydrase-regulated rice drought tolerance. Int. J. Mol. Sci. 2021, 22, 12119. [Google Scholar] [CrossRef]
  68. Muñoz-Vargas, M.A.; González-Gordo, S.; Palma, J.M.; Corpas, F.J. Inhibition of NADP-malic enzyme activity by H2 S and NO in sweet pepper (Capsicum annuum L.) fruits. Physiol. Plant. 2020, 168, 278–288. [Google Scholar]
  69. Riemenschneider, A.; Riedel, K.; Hoefgen, R.; Papenbrock, J.; Hesse, H. Impact of reduced O-acetylserine(thiol)lyase isoform contents on potato plant metabolism. Plant Physiol. 2005, 137, 892–900. [Google Scholar] [CrossRef] [Green Version]
  70. Alvarez, C.; Calo, L.; Romero, L.C.; García, I.; Gotor, C. An O-acetylserine(thiol)lyase homolog with L-cysteine desulfhydrase activity regulates cysteine homeostasis in Arabidopsis. Plant Physiol. 2010, 152, 656–669. [Google Scholar] [CrossRef] [Green Version]
  71. Willhardt, I.; Wiederanders, B. Activity staining of cystathionine-beta-synthetase and related enzymes. Anal. Biochem. 1975, 63, 263–266. [Google Scholar] [CrossRef]
  72. Calderwood, A.; Kopriva, S. Hydrogen sulfide in plants: From dissipation of excess sulfur to signaling molecule. Nitric Oxide 2014, 41, 72–78. [Google Scholar] [CrossRef] [PubMed]
  73. Zheng, J.L.; Hu, L.Y.; Hu, K.D.; Wu, J.; Yang, F.; Zhang, H. Hydrogen sulfide alleviates senescence of fresh-cut apple by regulating antioxidant defense system and senescence-related gene expression. HortScience 2016, 51, 152–158. [Google Scholar] [CrossRef] [Green Version]
  74. Ge, Y.; Hu, K.D.; Wang, S.S.; Hu, L.Y.; Chen, X.Y.; Li, Y.H. Hydrogen sulfide alleviates postharvest ripening and senescence of banana by antagonizing the effect of ethylene. PLoS ONE 2017, 12, e0180113. [Google Scholar] [CrossRef] [PubMed]
  75. Huo, J.; Huang, D.; Zhang, J.; Fang, H.; Wang, B.; Wang, C. Hydrogen sulfide: A gaseous molecule in postharvest freshness. Front. Plant Sci. 2018, 9, 1172. [Google Scholar] [CrossRef] [PubMed]
  76. Corpas, F.J.; Palma, J.M. H2S signaling in plants and applications in agriculture. J. Adv. Res. 2020, 24, 131–137. [Google Scholar] [CrossRef]
  77. Molinett, S.A.; Alfaro, J.F.; Sáez, F.A.; Elgueta, S.; Moya-León, M.A.; Figueroa, C.R. Postharvest treatment of hydrogen sulfide delays the softening of chilean strawberry fruit by downregulating the expression of key genes involved in pectin catabolism. Int. J. Mol. Sci. 2021, 22, 10008. [Google Scholar] [CrossRef]
  78. Siddiqui, M.W.; Homa, F.; Lata, D.; Mir, H.; Aftab, T.; Mishra, P. Hydrogen sulphide infiltration downregulates oxidative metabolism and extends postharvest life of banana. Plant Biol. 2021, in press. [CrossRef]
  79. Rose, P.; Moore, P.K.; Whiteman, M.; Zhu, Y.Z. An appraisal of developments in Allium sulfur chemistry: Expanding the pharmacopeia of garlic. Molecules 2019, 24, 4006. [Google Scholar] [CrossRef] [Green Version]
  80. Piragine, E.; Citi, V.; Lawson, K.; Calderone, V.; Martelli, A. Potential effects of natural H2S-donors in hypertension management. Biomolecules 2022, 12, 581. [Google Scholar] [CrossRef]
Ijms 23 05648 g001 550
Figure 1. Endogenous H2S detection in different plant species including (a) sweet pepper (Capsicum annuum L.) fruit at distinct ripening stages (fully green and fully red), (b) Broccoli (Brassica oleracea var. Itálica), (c) Allium species including garlic (Allium sativum L.) cloves, leek (Allium ampeloprasum var. porrum), welsh onion (Allium fistulosum), and purple onion (Allium cepa). H2S was detected using a micro sulfide ion electrode.
Figure 1. Endogenous H2S detection in different plant species including (a) sweet pepper (Capsicum annuum L.) fruit at distinct ripening stages (fully green and fully red), (b) Broccoli (Brassica oleracea var. Itálica), (c) Allium species including garlic (Allium sativum L.) cloves, leek (Allium ampeloprasum var. porrum), welsh onion (Allium fistulosum), and purple onion (Allium cepa). H2S was detected using a micro sulfide ion electrode.
Ijms 23 05648 g001
Ijms 23 05648 g002 550
Figure 2. H2S gas emission in different plant species including garlic (Allium sativum L.) cloves, leek (Allium ampeloprasum var. porrum), Welsh onion (Allium fistulosum), and purple onion (Allium cepa). H2S emission was recorded using an H2S sensor gas analyzer portable device. For each plant, 300 g of fresh material was used.
Figure 2. H2S gas emission in different plant species including garlic (Allium sativum L.) cloves, leek (Allium ampeloprasum var. porrum), Welsh onion (Allium fistulosum), and purple onion (Allium cepa). H2S emission was recorded using an H2S sensor gas analyzer portable device. For each plant, 300 g of fresh material was used.
Ijms 23 05648 g002
Ijms 23 05648 g003 550
Figure 3. L-Cysteine desulfhydrase (LCD) activity in different plant samples. (a) Spectrophotometric assay. (b) In-gel isozyme profile of LCD activity. Protein samples (74 µg protein per lane) were separated by non-denaturing polyacrylamide gel electrophoresis (PAGE; 8% acrylamide) and the LCD activity was detected by lead acetate staining (see M&M for details).
Figure 3. L-Cysteine desulfhydrase (LCD) activity in different plant samples. (a) Spectrophotometric assay. (b) In-gel isozyme profile of LCD activity. Protein samples (74 µg protein per lane) were separated by non-denaturing polyacrylamide gel electrophoresis (PAGE; 8% acrylamide) and the LCD activity was detected by lead acetate staining (see M&M for details).
Ijms 23 05648 g003
Publisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Share and Cite

MDPI and ACS Style

Muñoz-Vargas, M.A.; González-Gordo, S.; Palma, J.M.; Corpas, F.J. H2S in Horticultural Plants: Endogenous Detection by an Electrochemical Sensor, Emission by a Gas Detector, and Its Correlation with L-Cysteine Desulfhydrase (LCD) Activity. Int. J. Mol. Sci. 2022, 23, 5648. https://doi.org/10.3390/ijms23105648

AMA Style

Muñoz-Vargas MA, González-Gordo S, Palma JM, Corpas FJ. H2S in Horticultural Plants: Endogenous Detection by an Electrochemical Sensor, Emission by a Gas Detector, and Its Correlation with L-Cysteine Desulfhydrase (LCD) Activity. International Journal of Molecular Sciences. 2022; 23(10):5648. https://doi.org/10.3390/ijms23105648

Chicago/Turabian Style

Muñoz-Vargas, María A., Salvador González-Gordo, José M. Palma, and Francisco J. Corpas. 2022. "H2S in Horticultural Plants: Endogenous Detection by an Electrochemical Sensor, Emission by a Gas Detector, and Its Correlation with L-Cysteine Desulfhydrase (LCD) Activity" International Journal of Molecular Sciences 23, no. 10: 5648. https://doi.org/10.3390/ijms23105648

Note that from the first issue of 2016, this journal uses article numbers instead of page numbers. See further details here.

Article Metrics

Back to TopTop