Search

Hydrogen sulfide (H2S) is a signaling molecule that regulates essential plant processes. In this study, the role of H2S during drought was analysed, focusing on the underlying mechanism. Pretreatments with H2S before imposing drought on plants substantially improved the characteristic stressed phenotypes under drought and decreased the levels of typical biochemical stress markers such as anthocyanin, proline, and hydrogen peroxide. H2S also regulated drought-responsive genes and amino acid metabolism, and repressed drought-induced bulk autophagy and protein ubiquitination, demonstrating the protective effects of H2S pretreatment. Quantitative proteomic analysis identified 887 significantly different persulfidated proteins between control and drought stress plants. Bioinformatic analyses of the proteins more persulfidated in drought revealed that the most enriched biological processes were cellular response to oxidative stress and hydrogen peroxide catabolism. Protein degradation, abiotic stress responses, and the phenylpropanoid pathway were also highlighted, suggesting the importance of persulfidation in coping with drought-induced stress. Our findings emphasize the role of H2S as a promoter of enhanced tolerance to drought, enabling plants to respond more rapidly and efficiently. Furthermore, the main role of protein persulfidation in alleviating reactive oxygen species accumulation and balancing redox homeostasis under drought stress is highlighted.

Hydrogen sulfide (H

The healthy growth of plants is inseparable from the joint action of many metabolic regulators, particularly the signaling molecules. Hydrogen sulfide (H2S), which was considered toxic previously, has now been regarded as a burgeoning endogenous gaseous transmitter. H2S is involved in the whole life cycle of plants orderly, regulating essential plant processes such as seed germination, root growth, flowering, anti-aging of post-ripening by affecting corresponding gene expression and subsequent enzyme activities, as well as crosstalk with plant hormones and other signaling molecules. The research on H2S physiology entering an exponential exploration era, with significant progress has been made on many fronts of H2S function. However, the research on its molecular mechanism is still fragmented. In this review, we collect recent up-dates from physiological function to molecular biological mechanism of biological processes for H2S participating throughout plants’ life cycle, and present a brief outlook for future research.

Recent studies in mice and humans demonstrated the relevance of H2S synthesising enzymes, such as CTH, CBS, and MPST, in the physiology of adipose tissue and the differentiation of preadipocyte into adipocytes. Here, our objective was to investigate the combined role of CTH, CBS, and MPST in the preservation of adipocyte protein persulfidation and adipogenesis. Combined partial CTH, CBS, and MPST gene knockdown was achieved treating fully human adipocytes with siRNAs against these transcripts (siRNA_MIX). Adipocyte protein persulfidation was analyzed using label-free quantitative mass spectrometry coupled with a dimedone-switch method for protein labeling and purification. Proteomic analysis quantified 216 proteins with statistically different levels of persulfidation in KD cells compared to control adipocytes. In fully differentiated adipocytes, CBS and MPST mRNA and protein levels were abundant, while CTH expression was very low. It is noteworthy that siRNA_MIX administration resulted in a significant decrease in CBS and MPST expression, without impacting on CTH. The combined partial knockdown of the CBS and MPST genes resulted in reduced cellular sulfide levels in parallel to decreased expression of relevant genes for adipocyte biology, including adipogenesis, mitochondrial biogenesis, and lipogenesis, but increased proinflammatory- and senescence-related genes. It should be noted that the combined partial knockdown of CBS and MPST genes also led to a significant disruption in the persulfidation pattern of the adipocyte proteins. Although among the less persulfidated proteins, we identified several relevant proteins for adipocyte adipogenesis and function, among the most persulfidated, key mediators of adipocyte inflammation and dysfunction as well as some proteins that might play a positive role in adipogenesis were found. In conclusion, the current study indicates that the combined partial elimination of CBS and MPST (but not CTH) in adipocytes affects the expression of genes related to the maintenance of adipocyte function and promotes inflammation, possibly by altering the pattern of protein persulfidation in these cells, suggesting that these enzymes were required for the functional maintenance of adipocytes.

This paper presents a methodology for the localization of leaks in water distribution networks (WDNs) by means of the combination of a deep learning (DL) approach and a graph-based clustering technique. A data set for all possible leak locations is generated from pressure measurements and utilized to feed an image encoding process based on the Gramian Angular Field (GAF) technique, hence producing an equivalent data set of images. The pressure measurements are generated through the WDN simulation engine EPANET. To accomplish the training stage, the network is iteratively segmented into clusters using the Graph Agglomerative Clustering (GAC) method, and a deep learning neural network (DLNN) is trained to correctly indicate the leak location at one of the created clusters. The achieved neural networks tree can be traversed through its different branches depending on each classification result, until the final cluster is reached. Consequently, leaks can be located with a success rate that grows inversely to the size of the clusters. Due to the dependency of the latter on the number of clusters, which can be settled, the presented method is adaptable to the considered network features ( as e.g. dimensions, sensors placement and accuracy) and requisites (as e.g. localization area size).

Root hairs are specialized structures involved in water and nutrient uptake by plants. They elongate from epidermal cells following a complex developmental program. ß-cyanoalanine synthase (CAS), which is mainly involved in hydrogen cyanide (HCN) detoxification in Arabidopsis thaliana, plays a role in root hair elongation, as evidenced by the fact that cas-c1 mutants show a severe defect in root hair shape. In addition to root hairs, CAS C1 is expressed in the quiescent center and meristem. However, the cas-c1 mutation has no visible effect on either tissue, in both control and nutrient-deprivation conditions. To identify its role in root hair formation, we conducted single cell proteomics analysis by isolating root hair cells using Fluorescence-Activated Cell Sorting (FACS) from wild type and cas-c1 mutants. We also analyzed the presence of S-cyanylation, a protein post-translational modification (PTM) mediated by HCN and affecting cysteine residues and protein activity, in proteins of wild type and cas-c1 mutants. We found that several proteins involved in root hair development, related to the receptor kinase FERONIA signaling and to DNA methylation, are modified by this new post-translational modification.One sentence summaryArabidopsis root hair proteomics unveals that several proteins involved in root hair development are susceptible of modification by S-cyanylation.

Glucosinolates are antiherbivory chemical defense compounds in Arabidopsis (Arabidopsis thaliana). Specialist herbivores that feed on brassicaceous plants have evolved various mechanisms aimed at preventing the formation of toxic isothiocyanates. In contrast, generalist herbivores typically detoxify isothiocyanates through glutathione conjugation upon exposure. Here, we examined the response of an extreme generalist herbivore, the two-spotted spider mite Tetranychus urticae (Koch), to indole glucosinolates. Tetranychus urticae is a composite generalist whose individual populations have a restricted host range but have an ability to rapidly adapt to initially unfavorable plant hosts. Through comparative transcriptomic analysis of mite populations that have differential susceptibilities to Arabidopsis defenses, we identified β-cyanoalanine synthase of T. urticae (TuCAS), which encodes an enzyme with dual cysteine and β-cyanoalanine synthase activities. We combined Arabidopsis genetics, chemical complementation and mite reverse genetics to show that TuCAS is required for mite adaptation to Arabidopsis through its β-cyanoalanine synthase activity. Consistent with the β-cyanoalanine synthase role in detoxification of hydrogen cyanide (HCN), we discovered that upon mite herbivory, Arabidopsis plants release HCN. We further demonstrated that indole glucosinolates are sufficient for cyanide formation. Overall, our study uncovered Arabidopsis defenses that rely on indole glucosinolate-dependent cyanide for protection against mite herbivory. In response, Arabidopsis-adapted mites utilize the β-cyanoalanine synthase activity of TuCAS to counter cyanide toxicity, highlighting the mite’s ability to activate resistant traits that enable this extreme polyphagous herbivore to exploit cyanogenic host plants.

Alpha‐amylases are a large family of α,1‐4‐endo‐glycosyl hydrolases distributed in all kingdoms of life. The need for poly‐extremotolerant amylases encouraged their search in extreme environments, where archaea become ideal candidates to provide new enzymes that are able to work in the harsh conditions demanded in many industrial applications. In this study, a collection of haloarchaea isolated from Odiel saltern ponds in the southwest of Spain was screened for their amylase activity. The strain that exhibited the highest activity was selected and identified as Haloarcula sp. HS. We demonstrated the existence in both, cellular and extracellular extracts of the new strain, of functional α‐amylase activities, which showed to be moderately thermotolerant (optimum around 60 °C), extremely halotolerant (optimum over 25% NaCl), and calcium‐dependent. The tryptic digestion followed by HPLC‐MS/MS analysis of the partially purified cellular and extracellular extracts allowed to identify the sequence of three alpha‐amylases, which despite sharing a low sequence identity, exhibited high three‐dimensional structure homology, conserving the typical domains and most of the key consensus residues of α‐amylases. Moreover, we proved the potential of the extracellular α‐amylase from Haloarcula sp. HS to treat bakery wastes under high salinity conditions.

Hydrogen sulfide (H2S)-mediated signaling pathways regulate many physiological and pathophysiological processes in mammalian and plant systems. The molecular mechanism by which hydrogen sulfide exerts its action involves the posttranslational modification of cysteine residues to form a persulfidated thiol motif. We developed a comparative and label-free quantitative proteomic analysis approach for the detection of endogenous persulfidated proteins in N-starved Arabidopsis thaliana roots by using the tag-switch method. In this work, we identified 5214 unique proteins from root tissue that were persulfidated, 1674 of which were quantitatively analyzed and found to show altered persulfidation levels in vivo under N deprivation. These proteins represented almost 13% of the entire annotated proteome in Arabidopsis. Bioinformatic analysis revealed that persulfidated proteins were involved in a wide range of biological functions, regulating important processes such as primary metabolism, plant responses to stresses, growth and development, RNA translation and protein degradation. Quantitative mass spectrometry analysis allowed us to obtain a comprehensive view of hydrogen sulfide signaling via changes in the persulfidation levels of key protein targets involved in ubiquitin-dependent protein degradation and autophagy, among others

Aims: To investigate the impact of exogenous hydrogen sulfide (H2S) and its endogenous biosynthesis on human adipocytes and adipose tissue in the context of obesity and insulin resistance. Results: Experiments in human adipose tissue explants and in isolated preadipocytes demonstrated that exogenous H2S or the activation of endogenous H2S biosynthesis resulted in increased adipogenesis, insulin action, sirtuin deacetylase, and PPARγ transcriptional activity, whereas chemical inhibition and gene knockdown of each enzyme generating H2S (CTH, CBS, MPST) led to altered adipocyte differentiation, cellular senescence, and increased inflammation. In agreement with these experimental data, visceral and subcutaneous adipose tissue expression of H2S-synthesising enzymes was significantly reduced in morbidly obese subjects in association with attenuated adipogenesis and increased markers of adipose tissue inflammation and senescence. Interestingly, weight-loss interventions (including bariatric surgery or diet/exercise) improved the expression of H2S biosynthesis-related genes. In human preadipocytes, the expression of CTH, CBS, and MPST genes and H2S production were dramatically increased during adipocyte differentiation. More importantly, the adipocyte proteome exhibiting persulfidation was characterized, disclosing that different proteins involved in fatty acid and lipid metabolism, the citrate cycle, insulin signaling, several adipokines, and PPAR, experienced the most dramatic persulfidation (85–98%). Innovation: No previous studies investigated the impact of H2S on human adipose tissue. This study suggests that the potentiation of adipose tissue H2S biosynthesis is a possible therapeutic approach to improve adipose tissue dysfunction in patients with obesity and insulin resistance. Conclusion: Altogether, these data supported the relevance of H2S biosynthesis in the modulation of human adipocyte physiology.

This review focuses on recent advances relating to H2S signaling mechanisms, and highlights interconnections between H2S and H2O2 at the post-translational modification level and with ABA in stomatal movement., Hydrogen sulfide (H2S) is a signaling molecule that regulates critical processes and allows plants to adapt to adverse conditions. The molecular mechanism underlying H2S action relies on its chemical reactivity, and the most-well characterized mechanism is persulfidation, which involves the modification of protein thiol groups, resulting in the formation of persulfide groups. This modification causes a change of protein function, altering catalytic activity or intracellular location and inducing important physiological effects. H2S cannot react directly with thiols but instead can react with oxidized cysteine residues; therefore, H2O2 signaling through sulfenylation is required for persulfidation. A comparative study performed in this review reveals 82% identity between sulfenylome and persulfidome. With regard to abscisic acid (ABA) signaling, widespread evidence shows an interconnection between H2S and ABA in the plant response to environmental stress. Proteomic analyses have revealed persulfidation of several proteins involved in the ABA signaling network and have shown that persulfidation is triggered in response to ABA. In guard cells, a complex interaction of H2S and ABA signaling has also been described, and the persulfidation of specific signaling components seems to be the underlying mechanism.

With growing populations and pressing environmental problems, future economies will be increasingly plant-based. Now is the time to reimagine plant science as a critical component of fundamental science, agriculture, environmental stewardship, energy, technology and healthcare. This effort requires a conceptual and technological framework to identify and map all cell types, and to comprehensively annotate the localization and organization of molecules at cellular and tissue levels. This framework, called the Plant Cell Atlas (PCA), will be critical for understanding and engineering plant development, physiology and environmental responses. A workshop was convened to discuss the purpose and utility of such an initiative, resulting in a roadmap that acknowledges the current knowledge gaps and technical challenges, and underscores how the PCA initiative can help to overcome them., National Science Foundation 1916797 David W Ehrhardt, Kenneth D Birnbaum, Seung Yon Rhee; National Science Foundation 2052590 Seung Yon Rhee

Hydrogen sulfide (H2S) is a signaling molecule that regulates plant hormone and stress responses. The phytohormone abscisic acid (ABA) plays an important role in plant adaptation to unfavorable environmental conditions and induces the persulfidation of L-CYSTEINE DESULFHYDRASE1 (DES1) and the production of H2S in guard cells. However, it remains largely unclear how H2S and protein persulfidation participate in the relay of ABA signals. In this study, we discovered that ABSCISIC ACID INSENSITIVE 4 (ABI4) acts downstream of DES1 in the control of ABA responses in Arabidopsis. ABI4 undergoes persulfidation at Cys250 that is triggered in a time-dependent manner by ABA, and loss of DES1 function impairs this process. Cys250 and its persulfidation are essential for ABI4 function in the regulation of plant responses to ABA and the H2S donor NaHS during germination, seedling establishment, and stomatal closure, which are abolished in the ABI4Cys250Ala mutated variant. Introduction of the ABI4Cys250Ala variant into the abi4 des1 mutant did not rescue its hyposensitivity to ABA. Cys250 is critical for the binding of ABI4 to its cognate motif in the promoter of Mitogen-Activated Protein Kinase Kinase Kinase 18 (MAPKKK18), which propagates the MAPK signaling cascade induced by ABA. Furthermore, the DES1-mediated persulfidation of ABI4 enhances the transactivation activity of ABI4 toward MAPKKK18, and ABI4 can bind the DES1 promoter, forming a regulatory loop. Taken together, these findings advance our understanding of a post-translational regulatory mechanism and suggest that ABI4 functions as an integrator of ABA and MAPK signals through a process in which DES1-produced H2S persulfidates ABI4 at Cys250.

A control-oriented quality modeling approach is proposed for sewer networks, which can represent quality dynamics using simple equations in order to optimize pollution load from combined sewer overflows in large scale sewer network in real time. Total suspended solid has been selected as the quality indicator, regarding it is easy to be estimated through measuring turbidity and correlated with other quality indicators. The model equations are independent for different elements in sewer network, which allows a scalable usage. In order to ensure accuracy of the proposed models, a calibration procedure and a sensitivity analysis have been presented using data generated by virtual reality simulation. Afterwards, a quality-based model predictive control has been developed based on the proposed models. To validate effectiveness and efficiency of the modelling and optimization approaches, a pilot case, based on the Badalona sewer network in Spain is used. Application results under different scenarios show that the control-oriented modelling approach works properly to cope with quality dynamics in sewers. The quality-based optimization approach can provide strategies in reducing pollution loads in real time., The authors wish to acknowledge the support from the European Commission research grant of project LIFE EFFIDRAIN (LIFE14 ENV/ES/000860), from Spanish national project DEOCS (DPI2016-76493-C3-3-R) and the internal project TWINs from the CISC (Spanish State Research Agency) through the María de Maeztu Seal of Excellence to IRI (MDM-2016-0656). The authors also thank Aigües de Barcelona for their financial and technical support. Considering the fact that data, models and tools used in this research are confidential or commercial, we refer other researchers to contact SUEZ Spain Group (for access to detailed, simplified models; the Closed-loop Simulation Framework software; and the rain data) and Innovyze (for licenses to InfoWorks ICM). Also appreciate for the support from NWO (Netherlands Organisation for Scientific Research) Sectorplan for Beta and Technology Programme in Wageningen University

Los datos que se recolectan en escenarios de vulnerabilidad y riesgo, en el contexto de prevención de desastres naturales, son la base para consolidar información que permite disponer de los elementos para mitigar el impacto de los aspectos naturales que generan el riesgo; en tal sentido, cuando se estructura un sistema donde se capturan, almacenan y transforman los datos asociados con variables que miden o monitorean aspectos naturales particulares, se presenta la necesidad de asegurar esos datos para optimizar la fidelidad de la información y la titularidad de la misma. La investigación que se presenta se centra en el desarrollo de una plataforma con servicios sobre una interfaz web orientada al internet de las cosas, teniendo como sustento la garantía de la titularidad de los datos capturados a través de sensores conectados de forma remota y cuyos datos se procesan para la generación de reportes estadísticos descriptivos que facilitan la comprensión de las variables monitoreadas.

Gaseous molecules, such as hydrogen sulfide (H2 S) and nitric oxide (NO), are crucial players in cellular and (patho)physiological processes in biological systems. The biological functions of these gaseous molecules, which were first discovered and identified as gasotransmitters in animals, have received unprecedented attention from plant scientists in recent decades. Researchers have arrived at the consensus that H2 S is synthesized endogenously and serves as a signaling molecule throughout the plant life cycle. However, the mechanisms of H2 S action in redox biology is still largely unexplored. This review highlights what we currently know about the characteristics and biosynthesis of H2 S in plants. Additionally, we summarize the role of H2 S in plant resistance to abiotic stress. Moreover, we propose and discuss possible redox-dependent mechanisms by which H2 S regulates plant physiology.

Volatile compounds (VCs) emitted by the fungal phytopathogen Penicillium aurantiogriseum promote root growth and developmental changes in Arabidopsis. Here we characterized the metabolic and molecular responses of roots to fungal volatiles. Proteomic analyses revealed that these compounds reduce the levels of aquaporins, the iron carrier IRT1 and apoplastic peroxidases. Fungal VCs also increased the levels of enzymes involved in the production of mevalonate (MVA)‐derived isoprenoids, nitrogen assimilation and conversion of methionine to ethylene and cyanide. Consistently, fungal VC‐treated roots accumulated high levels of hydrogen peroxide (H2O2), MVA‐derived cytokinins, ethylene, cyanide and long‐distance nitrogen transport amino acids. qRT‐PCR analyses showed that many proteins differentially expressed by fungal VCs are encoded by VC non‐responsive genes. Expression patterns of hormone reporters and developmental characterization of mutants provided evidence for the involvement of cyanide scavenging and enhanced auxin, ethylene, cytokinin and H2O2 signaling in the root architecture changes promoted by fungal VCs. Our findings show that VCs from P. aurantiogriseum modify root metabolism and architecture, and improve nutrient and water use efficiencies through transcriptionally and non‐transcriptionally regulated proteome resetting mechanisms. Some of these mechanisms are subject to long‐distance regulation by photosynthesis and differ from those triggered by VCs emitted by beneficial microorganisms.

48 Pags.- 10 Figs. The definitive version, with suppl mat., is available at: http://www.plantcell.org/, Hydrogen sulfide is a signaling molecule that regulates essential processes in plants, such as autophagy. In Arabidopsis (Arabidopsis thaliana), hydrogen sulfide negatively regulates autophagy independently of reactive oxygen species via an unknown mechanism. Comparative and quantitative proteomic analysis was used to detect abscisic acid-triggered persulfidation that reveals a main role in the control of autophagy mediated by the autophagy-related (ATG) Cys protease AtATG4a. This protease undergoes specific persulfidation of Cys170 that is a part of the characteristic catalytic Cys-His-Asp triad of Cys proteases. Regulation of the ATG4 activity by persulfidation was tested in a heterologous assay using the Chlamydomonas reinhardtii CrATG8 protein as a substrate. Sulfide significantly and reversibly inactivates AtATG4a. The biological significance of the reversible inhibition of the ATG4 by sulfide is supported by the results obtained in Arabidopsis leaves under basal and autophagy-activating conditions. A significant increase in the overall ATG4 proteolytic activity in Arabidopsis was detected under nitrogen starvation and osmotic stress and can be inhibited by sulfide. Therefore, the data strongly suggest that the negative regulation of autophagy by sulfide is mediated by specific persulfidation of the ATG4 protease., This work was supported in part by the European Regional Development Fund through the Ministerio de Economía y Competitividad and the Agencia Estatal de Investigación (grant nos. BFU2015-68216-P and PGC2018-099048- B-I00 to J.L.C.; grant nos. BIO2015-74432-JIN and PID2019- 110080GB-I00 to M.E.P.-P.; grant nos. BIO2016-76633-P and PID2019- 109785GB-I00; and Junta de Andalucía grant no. P18-RT-3154 to C.G.)., the Marie Skłodowska-Curie Grant Agreement (grant no. 834120 to Á.A.), Government of Aragón-FEDER (grant no. E35_17R to I.Y.), and the Ministerio de Economia y Competitividad through the program of “Formación de Personal Investigador” (to A.M.L.-M. and A.J.-F.).

Recent studies have demonstrated that hydrogen sulfide (H2 S) produced through the activity of l-cysteine desulfhydrase (DES1) is an important gaseous signaling molecule in plants that could participate in abscisic acid (ABA)-induced stomatal closure. However, the coupling of the DES1/H2 S signaling pathways to guard cell movement has not been thoroughly elucidated. The results presented here provide genetic evidence for a physiologically relevant signaling pathway that governs guard cell in situ DES1/H2 S function in stomatal closure. We discovered that ABA-activated DES1 produces H2 S in guard cells. The impaired guard cell ABA phenotype of the des1 mutant can be fully complemented when DES1/H2 S function has been specifically rescued in guard cells and epidermal cells, but not mesophyll cells. This research further characterized DES1/H2 S function in the regulation of LONG HYPOCOTYL1 (HY1, a member of the heme oxygenase family) signaling. ABA-induced DES1 expression and H2 S production are hyper-activated in the hy1 mutant, both of which can be fully abolished by the addition of H2 S scavenger. Impaired guard cell ABA phenotype of des1/hy1 can be restored by H2 S donors. Taken together, this research indicated that guard cell in situ DES1 function is involved in ABA-induced stomatal closure, which also acts as a pivotal hub in regulating HY1 signaling.

Hydrogen sulfide (H(2)S) is a gaseous signaling molecule that regulates diverse cellular signaling pathways through persulfidation, which involves the post-translational modification of specific Cys residues to form persulfides. However, the mechanisms that underlie this important redox-based modification remain poorly understood in higher plants. We have, therefore, analyzed how protein persulfidation acts as a specific and reversible signaling mechanism during the abscisic acid (ABA) response in Arabidopsis (Arabidopsis thaliana). Here we show that ABA stimulates the persulfidation of l-CYSTEINE DESULFHYDRASE1, an important endogenous H(2)S enzyme, at Cys44 and Cys205 in a redox-dependent manner. Moreover, sustainable H(2)S accumulation drives persulfidation of the NADPH oxidase RESPIRATORY BURST OXIDASE HOMOLOG PROTEIN D (RBOHD) at Cys825 and Cys890, enhancing its ability to produce reactive oxygen species. Physiologically, s-persulfidation-induced RBOHD activity is relevant to ABA-induced stomatal closure. Together, these processes form a negative feedback loop that fine-tunes guard cell redox homeostasis and ABA signaling. These findings not only expand our current knowledge of H(2)S function in the context of guard cell ABA signaling, but also demonstrate the presence of a rapid signal integration mechanism involving specific and reversible redox-based post-translational modifications that occur in response to changing environmental conditions.

Hydrogen sulfide (H2S) has always been considered toxic, but a huge number of articles published more recently showed the beneficial biochemical properties of its endogenous production throughout all regna. In this review, the participation of H2S in many physiological and pathological processes in animals is described, and its importance as a signaling molecule in plant systems is underlined from an evolutionary point of view. H2S quantification methods are summarized and persulfidation is described as the underlying mechanism of action in plants, animals and bacteria. This review aims to highlight the importance of its crosstalk with other signaling molecules and its fine regulation for the proper function of the cell and its survival., This work has received funding from the European Union’s Horizon 2020 research and innovation programme under the Marie Skłodowska-Curie grant agreement No. 834120.

Hydrogen cyanide (HCN) is coproduced with ethylene in plant cells and is primarily enzymatically detoxified by the mitochondrial β-CYANOALANINE SYNTHASE (CAS-C1). Permanent or transient depletion of CAS-C1 activity in Arabidopsis (Arabidopsis thaliana) results in physiological alterations in the plant that suggest that HCN acts as a gasotransmitter molecule. Label-free quantitative proteomic analysis of mitochondrially enriched samples isolated from the wild type and cas-c1 mutant revealed significant changes in protein content, identifying 451 proteins that are absent or less abundant in cas-c1 and 353 proteins that are only present or more abundant in cas-c1 Gene ontology classification of these proteins identified proteomic changes that explain the root hairless phenotype and the altered immune response observed in the cas-c1 mutant. The mechanism of action of cyanide as a signaling molecule was addressed using two proteomic approaches aimed at identifying the S-cyanylation of Cys as a posttranslational modification of proteins. Both the 2-imino-thiazolidine chemical method and the direct untargeted analysis of proteins using liquid chromatography-tandem mass spectrometry identified a set of 163 proteins susceptible to S-cyanylation that included SEDOHEPTULOSE 1,7-BISPHOSPHATASE (SBPase), the PEPTIDYL-PROLYL CIS-TRANS ISOMERASE 20-3 (CYP20-3), and ENOLASE2 (ENO2). In vitro analysis of these enzymes showed that S-cyanylation of SBPase Cys74, CYP20-3 Cys259, and ENO2 Cys346 residues affected their enzymatic activity. Gene Ontology classification and protein-protein interaction cluster analysis showed that S-cyanylation is involved in the regulation of primary metabolic pathways, such as glycolysis, and the Calvin and S-adenosyl-Met cycles.

In Arabidopsis thaliana, cyanide is produced concomitantly with ethylene biosynthesis and is mainly detoxified by the ß-cyanoalanine synthase CAS-C1. In roots, CAS-C1 activity is essential to maintain a low level of cyanide for proper root hair development. Root hair elongation relies on polarized cell expansion at the growing tip, and we have observed that CAS-C1 locates in mitochondria and accumulates in root hair tips during root hair elongation, as shown by observing the fluorescence in plants transformed with the translational construct ProC1:CASC1-GFP, containing the complete CAS-C1 gene fused to GFP. Mutants in the SUPERCENTIPEDE (SCN1) gene, that regulate the NADPH oxidase RHD2/AtrbohC, are affected at the very early steps of the development of root hair that do not elongate and do not show a preferential localization of the GFP accumulation in the tips of the root hair primordia. Root hairs of mutants in CAS-C1 or RHD2/AtrbohC, which catalyzes the generation of ROS and the Ca2+ gradient, correctly start to grow out but they do not elongate either. Genetic crosses between the cas-c1 mutant and scn1 or rhd2 mutants were performed and the detail phenotypic and molecular characterization of the double mutants demonstrate that scn1 mutation is epistatic to cas-c1 and cas-c1 is epistatic to rhd2 mutation, indicating that CAS-C1 acts in early steps of the root hair development process. Moreover, our results show that the role of CAS-C1 in root hair elongation is independent of H2O2 production and of a direct NADPH oxidase inhibition by cyanide

Peroxisomes, which are ubiquitous organelles in all eukaryotes, are highly dynamic organelles that are essential for development and stress responses. Plant peroxisomes are involved in major metabolic pathways, such as fatty acid β-oxidation, photorespiration, ureide and polyamine metabolism, in the biosynthesis of jasmonic, indolacetic, and salicylic acid hormones, as well as in signaling molecules such as reactive oxygen and nitrogen species (ROS/RNS). Peroxisomes are involved in the perception of environmental changes, which is a complex process involving the regulation of gene expression and protein functionality by protein post-translational modifications (PTMs). Although there has been a growing interest in individual PTMs in peroxisomes over the last ten years, their role and cross-talk in the whole peroxisomal proteome remain unclear. This review provides up-to-date information on the function and crosstalk of the main peroxisomal PTMs. Analysis of whole peroxisomal proteomes shows that a very large number of peroxisomal proteins are targeted by multiple PTMs, which affect redox balance, photorespiration, the glyoxylate cycle, and lipid metabolism. This multilevel PTM regulation could boost the plasticity of peroxisomes and their capacity to regulate metabolism in response to environmental changes., This study was co-funded by ERDF (UE) and the Spanish Ministry of Science, Innovation and Universities grants PGC2018-098372-B-I00 and BIO2016-76633-P.

Two cysteine metabolism-related molecules, hydrogen sulfide and hydrogen cyanide, which are considered toxic, have now been considered as signaling molecules. Hydrogen sulfide is produced in chloroplasts through the activity of sulfite reductase and in the cytosol and mitochondria by the action of sulfide-generating enzymes, and regulates/affects essential plant processes such as plant adaptation, development, photosynthesis, autophagy, and stomatal movement, where interplay with other signaling molecules occurs. The mechanism of action of sulfide, which modifies protein cysteine thiols to form persulfides, is related to its chemical features. This post-translational modification, called persulfidation, could play a protective role for thiols against oxidative damage. Hydrogen cyanide is produced during the biosynthesis of ethylene and camalexin in non-cyanogenic plants, and is detoxified by the action of sulfur-related enzymes. Cyanide functions include the breaking of seed dormancy, modifying the plant responses to biotic stress, and inhibition of root hair elongation. The mode of action of cyanide is under investigation, although it has recently been demonstrated to perform post-translational modification of protein cysteine thiols to form thiocyanate, a process called S-cyanylation. Therefore, the signaling roles of sulfide and most probably of cyanide are performed through the modification of specific cysteine residues, altering protein functions.

Hydrogen sulfide (H2S) is a gaseous signaling molecule that regulates diverse cellular signaling through post translational modifications (PTM) of specific cysteine residues to form persulfides. However, this important redox-based PTM remains poorly understood. We have therefore analyzed how protein persulfidation is utilized as a specific, reversible signaling mechanism. Here we show the reciprocal control between DES1-produced H2S and guard cell autonomous ABA synthesis in regulation of stomatal water loss. ABA stimulates DES1 itself persulfidation at Cys44/205 in a redox-dependent manner. Moreover, sustained H2S accumulation drives persulfidation of the RBOHD protein at Cys825/890, enhancing ROS-production. Together, these processes form a negative feedback loop that fine-tunes guard cell redox homeostasis and ABA signaling. These findings not only expand our current knowledge of H2S function in the context of ABA metabolism and signaling, but also demonstrate the presence of a rapid signal integration mechanism with regard to specific and reversible redox-based PTMs in response to changing environmental conditions.

Cadmium is one of the most common heavy metals in contaminated aquatic environments and one of the most toxic contaminants for phytoplankton. Nevertheless, there are not enough studies focused on the effect of this metal in algae. Through a proteomic approach, this work shows how Cd can alter the growth, cell morphology and metabolism of the microalga Chlorella sorokiniana. Using the sequential window acquisition of all theoretical fragment ion spectra mass spectrometry (SWATH-MS), we concluded that exposure of Chlorella sorokiniana to 250 μM Cd2+ for 40 h caused downregulation of different metabolic pathways, such as photosynthesis, oxidative phosphorylation, glycolysis, TCA cycle and ribosomal proteins biosynthesis. However, photorespiration, antioxidant enzymes, gluconeogenesis, starch catabolism, and biosynthesis of glutamate, cysteine, glycine and serine were upregulated, under the same conditions. Finally, exposure to Cd also led to changes in the metabolism of carotenoids and lipids. In addition, the high tolerance of Chlorella sorokiniana to Cd points to this microalga as a potential microorganism to be used in bioremediation processes., This work was supported by European governments (INTERREG VA- POCTEP- 2014–2020; 0055_ALGARED_PLUS_5_E) and the European Regional Development Fund through the Agencia Estatal de Inves-tigaci ́on grants (PID 2019-109785 GB-I00 and PID 2019-110438RB- C22). We want to thank Dr. Rocío Rodríguez for the proteomics service.

In non-cyanogenic plants, cyanide is produced during ethylene biosynthesis and is mainly detoxified by the ß-cyanoalanine synthase CAS-C1. Arabidopsis plants lacking CAS-C1 show abnormal root hairs, which stop growing at early stages. Root hair elongates by polarized cell expansion at the tip, and we have observed that CAS-C1-driven GFP fluorescence locates in mitochondria and accumulates in root hair tips during root hair elongation. Genetic crosses have been performed between cas-c1 plants and scn1-1 mutants, defective in the SCN1 protein that regulates the NADPH oxidase RHD2/AtrbohC, and between cas-c1 and rhd2-1, defective in the NADPH oxidase necessary for the generation of ROS and the Ca2+ gradient necessary for root hair elongation. The phenotypic and molecular analysis of these crosses indicates that cas-c1 is hypostatic to scn1-1 and epistatic to rhd2-1. Furthermore, the action of cyanide in root hair development is independent of ROS and of direct NADPH oxidase inhibition by cyanide.

Objective To evaluate the efficacy of a physiotherapy program for relieving symptoms of primary dysmenorrhea among Mexican women. Study design This was a single-center, prospective, experimental, parallel group, randomized controlled trial. This cross-sectional study was performed at the Universidad Politecnica de Pachuca, Hidalgo, Mexico. Female patients with primary dysmenorrhea, age of 18–22 years; pain intensity from 4 to 10 cm on a Visual Analogue Scale (VAS); and sedentary lifestyle were included. The patients were then randomized to receive a physiotherapy program for three months or to no intervention program. The physiotherapy program consisted of overall stretching, specific stretches, Kegel exercises, jogging, and relaxation exercises. Patient evaluations of symptomatology and pain intensity were recorded basally and throughout for three menstrual periods. The data were entered into a computerized database for descriptive and inferential statistical analyses. Results A per-protocol population of eighty three women with a mean age of 20.2 ± 1.8 years underwent the physiotherapy program, and seventy seven participants with a mean age of 20.4 ± 1.2 years received no treatment. The participant assessments of pain on the VAS during the second and the third menstrual cycles demonstrated a significant reduction in the treatment group ( p Conclusions The results showed that strengthening, stretching and muscle relaxation techniques, in addition to jogging, are effective for reducing dysmenorrheic symptoms when they are regularly performed.

Hydrogen sulfide is a highly reactive molecule that is currently accepted as a signaling compound. This molecule is as important as carbon monoxide in mammals and hydrogen peroxide in plants, as well as nitric oxide in both eukaryotic systems. Although many studies have been conducted on the physiological effects of hydrogen sulfide, the underlying mechanisms are poorly understood. One of the proposed mechanisms involves the posttranslational modification of protein cysteine residues, a process called S-sulfhydration. In this work, a modified biotin switch method was used for the detection of Arabidopsis (Arabidopsis thaliana) proteins modified by S-sulfhydration under physiological conditions. The presence of an S-sulfhydration-modified cysteine residue on cytosolic ascorbate peroxidase was demonstrated using liquid chromatography-tandem mass spectrometry analysis, and a total of 106 S-sulfhydrated proteins were identified. Immunoblot and enzyme activity analyses of some of these proteins showed that the sulfide added through S-sulfhydration reversibly regulates the functions of plant proteins in a manner similar to that described in mammalian systems.