Revealing on hydrogen sulfide and nitric oxide signals co-ordination for plant growth under stress conditions

In the recent times, plants are facing certain types of environmental stresses, which give rise to formation of reactive oxygen species (ROS) such as hydroxyl radicals, hydrogen peroxides, superoxide anions and so on. These are required by the plants at low concentrations for signal transduction and at high concentrations, they repress plant root growth. Apart from the ROS activities, hydrogen sulfide (H₂S) and nitric oxide (NO) have major contributions in regulating growth and developmental processes in plants, as they also play key roles as signaling molecules and act as chief plant immune defense mechanisms against various biotic as well as abiotic stresses. H₂S and NO are the two pivotal gaseous messengers involved in growth, germination and improved tolerance in plants under stressed and non-stress conditions. H₂S and NO mediate cell signaling in plants as a response to several abiotic stresses like temperature, heavy metal exposure, water and salinity. They alter gene expression levels to induce the synthesis of antioxidant enzymes, osmolytes and also trigger their interactions with each other. However, research has been limited to only cross adaptations and signal transductions. Understanding the change and mechanism of H₂S and NO mediated cell signaling will broaden our knowledge on the various biochemical changes that occur in plant cells related to different stresses. A clear understanding of these molecules in various environmental stresses would help to confer biotechnological applications to protect plants against abiotic stresses and to improve crop productivity.     

Regulation of physiological aspects in plants by hydrogen sulfide and nitric oxide under challenging environment

Plants are exposed to a plethora of abiotic stresses such as drought, salinity, heavy metal and temperature stresses at different stages of their life cycle, from germination to seedling till the reproductive phase. As protective mechanisms, plants release signaling molecules that initiate a cascade of stress-signaling events, leading either to programmed cell death or plant acclimation. Hydrogen sulfide (H₂S) and nitric oxide (NO) are considered as new ‘gasotransmitter’ molecules that play key roles in regulating gene expression, posttranslational modification (PTM), as well as cross-talk with other hormones. Although the exact role of NO in plants remains unclear and is species dependent, various studies have suggested a positive correlation between NO accumulation and environmental stress in plants. These molecules are also involved in a large array of stress responses and act synergistically or antagonistically as signaling components, depending on their respective concentration. This study provides a comprehensive update on the signaling interplay between H₂S and NO in the regulation of various physiological processes under multiple abiotic stresses, modes of action and effects of exogenous application of these two molecules under drought, salt, heat and heavy metal stresses. However, the complete picture of the signaling cascades mediated by H₂S and NO is still elusive. Recent researches indicate that during certain plant processes, such as stomatal closure, H₂S could act upstream of NO signaling or downstream of NO in response to abiotic stresses by improving antioxidant activity in most plant species. In addition, PTMs of antioxidative pathways by these two molecules are also discussed.     

Structural evolution and function of stress associated proteins in regulating biotic and abiotic stress responses in plants

Adverse environmental conditions greatly influence crop production every year and threaten food security. Plants have a range of signaling networks to combat these stresses, in which several stress-responsive genes and regulatory proteins function together. One such important family of proteins, the Stress Associated Protein (SAP) family, has been identified as a novel regulator of multiple stresses. The SAPs possess a characteristic N-terminal A20 zinc-finger domain combined with either AN1 or C₂H₂ at the C-terminus. SAPs provide tolerance against various abiotic stresses, including cold, salt, drought, heavy metal, and wounding. The majority of SAPs are stress-inducible and have a function in conferring stress tolerance in transgenics. The role of SAPs in regulating biotic stress responses is a newly emerging field among researchers. SAPs interact with many other proteins to execute their functions; however, the detailed mechanism of these interactions needs to be elucidated. In this context, the present review provides a detailed view of the evolution and functions of SAPs in plants. The involvement in crosstalk between abiotic and biotic stress signaling pathways makes SAPs ideal targets to develop crops with tolerance against multiple stresses without any yield penalty. Altogether, we provide current knowledge on SAPs for investigating their role in stress response, which can further be exploited to develop climate-resilient crops through transgene-based, breeding-mediated, or genome-editing approaches.

     

Stress responsive gene regulation in relation to hydrogen sulfide in plants under abiotic stress

Plants often face a variety of abiotic stresses, which affects them negatively and lead to yield loss. The antioxidant system efficiently removes excessive reactive oxygen species and maintains redox homeostasis in plants. With better understanding of these protective mechanisms, recently the concept of hydrogen sulfide (H₂S) and its role in cell signaling has become the center of attention. H₂S has been recognized as a third gasotransmitter and a potent regulator of growth and development processes such as germination, maturation, senescence and defense mechanism in plants. Because of its gaseous nature, H₂S can diffuse to different part of the cells and balance the antioxidant pools by supplying sulfur to cells. H₂S showed tolerance against a plethora of adverse environmental conditions like drought, salt, high temperature, cold, heavy metals and flood via changing in level of osmolytes, malonaldialdehyde, Na⁺/K⁺ uptake, activities of H₂S biosynthesis and antioxidative enzymes. It also promotes cross adaptation through persulfidation. H₂S along with calcium, methylglyoxal and nitric oxide, and their cross talk induces the expression of mitogen activated protein kinases as well as other genes in response to stress. Therefore, it is sensible to evaluate and explore the stress responsive genes involved in H₂S regulated homeostasis and stress tolerance. The current article is aimed to summarize the recent updates on H₂S-mediated gene regulation in special reference to abiotic stress tolerance mechanism, and cross adaptation in plants. Moreover, new insights into the H₂S-associated signal transduction pathway have also been explored.     

Insights into the Role of Gasotransmitters Mediating Salt Stress Responses in Plants

Salinity stress is one of the most significant global issues that negatively affect plant growth and development. Modern agricultural practices have expanded the destructive effects of salinity stress, affecting plants through immediate osmotic stress, followed by a slow onset of ionic or hyper-osmotic stress. Plants alteration and resistance to salinity stress involve complex physiological, biochemical, and molecular systems to maintain homeostasis. As of late, the investigation of gaseous molecules in plants has attained much consideration, particularly for abiotic stress. Abiotic stresses generally initiate gasotransmitter (GT) generation in plants. In the interim, these GTs enhance the accumulation and activities of few antioxidant molecules, check the destructiveness of reactive oxygen species (ROS), and improve plant resilience under different stress conditions. The current review presented the role of gaseous molecules in plants under salinity stress, which include nitric oxide (·NO), hydrogen sulfide (H₂S), hydrogen gas (H₂), carbon monoxide (CO), methane (CH₄), and the only gaseous phytohormone ethylene. Further, we highlighted the underlying molecular mechanisms of the gasotransmitter signaling and cross-talks in salinity stress. Also, we presented a general update on the inclusion of GT in salt stress response, including the research gaps and its applications in the advancement of salinity-resistant plants.

     

Protective role of nitric oxide against arsenic-induced damages in germinating mung bean seeds

Nitric oxide (NO) is a bioactive gaseous, multifunctional molecule playing a central role and mediating a variety of physiological processes and responses to biotic and abiotic stresses including heavy metals. The present study investigated whether NO applied exogenously as sodium nitroprusside (SNP) has a protective role against arsenic (As) toxicity (applied as sodium arsenate) in Vigna radiata (mung bean) germinating seeds. Treatment with 75???M SNP significantly improved mung bean seed germination, growth, and decreased the As-accumulation. Furthermore, As-induced oxidative stress measured in terms of malondialdehyde and H₂O₂ contents was lesser upon supplementation of SNP indicating a reactive oxygen species scavenging activity of NO. In addition, supplementation of SNP markedly decreased the activity of superoxide dismutase and stimulated catalase, ??-amylase, protease and slightly changed the H⁺-ATPase activity.     

Overexpression of Rat Neurons Nitric Oxide Synthase in Rice Enhances Drought and Salt Tolerance

Nitric oxide (NO) has been shown to play an important role in the plant response to biotic and abiotic stresses in Arabidopsis mutants with lower or higher levels of endogenous NO. The exogenous application of NO donors or scavengers has also suggested an important role for NO in plant defense against environmental stress. In this study, rice plants under drought and high salinity conditions showed increased nitric oxide synthase (NOS) activity and NO levels. Overexpression of rat neuronal NO synthase (nNOS) in rice increased both NOS activity and NO accumulation, resulting in improved tolerance of the transgenic plants to both drought and salt stresses. nNOS-overexpressing plants exhibited stronger water-holding capability, higher proline accumulation, less lipid peroxidation and reduced electrolyte leakage under drought and salt conditions than wild rice. Moreover, nNOS-overexpressing plants accumulated less H₂O₂, due to the observed up-regulation of OsCATA, OsCATB and OsPOX1. In agreement, the activities of CAT and POX were higher in transgenic rice than wild type. Additionally, the expression of six tested stress-responsive genes including OsDREB2A, OsDREB2B, OsSNAC1, OsSNAC2, OsLEA3 and OsRD29A, in nNOS-overexpressing plants was higher than that in the wild type under drought and high salinity conditions. Taken together, our results suggest that nNOS overexpression suppresses the stress-enhanced electrolyte leakage, lipid peroxidation and H₂O₂ accumulation, and promotes proline accumulation and the expression of stress-responsive genes under stress conditions, thereby promoting increased tolerance to drought and salt stresses.     

The role of mitochondrial reactive oxygen species,NO and H2S in ischaemia/reperfusion injury and cardioprotection

Redox signalling in mitochondria plays an important role in myocardial ischaemia/reperfusion (I/R) injury and in cardioprotection. Reactive oxygen and nitrogen species (ROS/RNS) modify cellular structures and functions by means of covalent changes in proteins including among others S‐nitros(yl)ation by nitric oxide (NO) and its derivatives, and S‐sulphydration by hydrogen sulphide (H₂S). Many enzymes are involved in the mitochondrial formation and handling of ROS, NO and H₂S under physiological and pathological conditions. In particular, the balance between formation and removal of reactive species is impaired during I/R favouring their accumulation. Therefore, various interventions aimed at decreasing mitochondrial ROS accumulation have been developed and have shown cardioprotective effects in experimental settings. However, ROS, NO and H₂S play also a role in endogenous cardioprotection, as in the case of ischaemic pre‐conditioning, so that preventing their increase might hamper self‐defence mechanisms. The aim of the present review was to provide a critical analysis of formation and role of reactive species, NO and H₂S in mitochondria, with a special emphasis on mechanisms of injury and protection that determine the fate of hearts subjected to I/R. The elucidation of the signalling pathways of ROS, NO and H₂S is likely to reveal novel molecular targets for cardioprotection that could be modulated by pharmacological agents to prevent I/R injury.     

Expression of apoplastically secreted tobacco osmotin in cotton confers drought tolerance

Osmotin or osmotin-like proteins have been shown to be induced in several plant species in response to various types of biotic and abiotic challenges. The protein is generally believed to be involved in protecting the plant against these stresses. Although some understanding of the possible mechanism underlying the defense function of osmotin against biotic stresses is beginning to emerge, its role in abiotic stress response is far from clear. We have transformed cotton plants with a tobacco-osmotin gene, lacking the sequence encoding its 20 amino acid-long, C-terminal vacuolar-sorting motif, under the control of CaMV 35S promoter. Apoplastic secretion of the recombinant protein was confirmed and the plants were evaluated for their ability to tolerate drought conditions. Under polyethylene glycol-mediated water stress, the osmotin-expressing seedlings showed better growth performance. The transformants showed a slower rate of wilting during drought and faster recovery following the termination of dry conditions in a greenhouse setting. During drought, the leaves from transgenic plants had higher relative water content and proline levels, while showing reduced H₂O₂ levels, lipid peroxidation and electrolyte leakage. Importantly, following a series of dry periods, the osmotin transformants performed better in terms of most growth and developmental parameters tested. Most relevant, the fiber yield of transgenic plants did not suffer as much as that of their non-transgenic counterparts under drought conditions. The results provide direct support for a protective role of osmotin in cotton plants experiencing water stress and suggest a possible way to achieve tolerance to drought conditions by means of genetic engineering. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Influence of exogenously applied nitric oxide on strawberry (Fragaria × ananassa) plants grown under iron deficiency and/or saline stress

A study was carried out to assess the protective effects of exogenously applied nitric oxide (NO) in the form of its donor sodium nitroprusside (SNP) to strawberry seedlings (Fragaria × ananassa cv. Camarosa) grown under iron deficiency (ID), salinity stress or combination of both. The experimental design contained control, 0.1 mM FeSO₄ (ID, Fe deficiency); 50 mM NaCl (S, Salinity) and ID + S. Plants were sprayed with 0.1 mM SNP or 0.1 mM sodium ferrocyanide, an analogue of SNP containing no NO. The deleterious effects of ID + S treatments on plant fresh and dry matters, total chlorophyll and chlorophyll fluorescence were more striking than those caused by the ID or S treatment alone. Furthermore, combination of salinity and iron stress exacerbated electrolyte leakage (EL) and the levels of malondialdehyde (MDA) and hydrogen peroxide (H₂O₂) in plant leaves compared to those in plants grown with either of the single stresses. NO treatment effectively reduced EL, MDA and H₂O₂ in plants grown under stress conditions applied singly or in combination. Salt stress alone and with ID reduced the superoxide dismutase (EC1.15.1.1) and catalase (EC 1.11.1.6) activities but increased that of POD (EC 1.17.1.7). Exogenously applied NO led to significant changes in antioxidant enzyme activities in either ID or S than those by ID+S. Overall, exogenously applied NO was more effective in mitigating the stress‐induced adverse effects on the strawberry plants exposed to a single stress than those due to the combination of both stresses.     

Hydrogen sulfide,a signaling molecule in plant stress responses

Gaseous molecules, such as hydrogen sulfide(H_2S)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 H_2S is synthesized endogenously and serves as a signaling molecule throughout the plant life cycle.However, the mechanisms of H_2S action in redox biology is still largely unexplored. This review highlights what we currently know about the characteristics and biosynthesis of H_2S in plants. Additionally,we summarize the role of H_2S in plant resistance to abiotic stress. Moreover, we propose and discuss possible redox-dependent mechanisms by which H_2S regulates plant physiology.     

Hydrogen sulfide interacts with calcium signaling to enhance the chromium tolerance in Setaria italica

The oscillation of intracellular calcium (Ca²⁺) concentration is a primary event in numerous biological processes in plants, including stress response. Hydrogen sulfide (H₂S), an emerging gasotransmitter, was found to have positive effects in plants responding to chromium (Cr⁶⁺) stress through interacting with Ca²⁺ signaling. While Ca²⁺ resemblances H₂S in mediating biotic and abiotic stresses, crosstalk between the two pathways remains unclear. In this study, Ca²⁺ signaling interacted with H₂S to produce a complex physiological response, which enhanced the Cr⁶⁺ tolerance in foxtail millet (Setaria italica). Results indicate that Cr⁶⁺ stress activated endogenous H₂S synthesis as well as Ca²⁺ signaling. Moreover, toxic symptoms caused by Cr⁶⁺ stress were strongly moderated by 50 μM H₂S and 20 mM Ca²⁺. Conversely, treatments with H₂S synthesis inhibitor and Ca²⁺ chelators prior to Cr⁶⁺-exposure aggravated these toxic symptoms. Interestingly, Ca²⁺ upregulated expression of two important factors in metal metabolism, MT3A and PCS, which participated in the biosynthesis of heavy metal chelators, in a H₂S-dependent manner to cope with Cr⁶⁺ stress. These findings also suggest that the H₂S dependent pathway is a component of the Ca²⁺ activating antioxidant system and H₂S partially contributes Ca²⁺-activating antioxidant system.     

Enhanced thermo-tolerance in transgenic potato (Solanum tuberosum L.) overexpressing hydrogen peroxide-producing germin-like protein (GLP)

Germins and germin-like proteins (GLPs) were reported to participate in plant response to biotic and abiotic stresses involving hydrogen peroxide (H₂O₂) production, but their role in mitigating heat stress is poorly understood. Here, we investigated the ability of a Solanum tuberosum L. GLP (StGLP) gene isolated from the yeast cDNA library generated from heat-stressed potato plants and characterized its role in generating innate and/or acquired thermo-tolerance to potato via genetic transformation. The transgenic plants exhibited enhanced tolerance to gradual heat stress (GHS) compared with sudden heat shock (SHS) in terms of maximal cell viability, minimal ion leakage and reduced chlorophyll breakdown. Further, three StGLP transgenic lines (G9, G12 and G15) exhibited enhanced production of H₂O₂, which was either reduced or blocked by inhibitors of H₂O₂ under normal and heat stress conditions. This tolerance was mediated by up-regulation of antioxidant enzymes (catalase, ascorbate peroxidase and glutathione reductase) and other heat stress-responsive genes (StHSP70, StHSP20 and StHSP90) in transgenic potato plants. These results demonstrate that H₂O₂ produced by over-expression of StGLP in transgenic potato plants triggered the reactive oxygen species (ROS) scavenging signaling pathways controlling antioxidant and heat stress-responsive genes in these plants imparting tolerance to heat stress.     

Integrative roles of nitric oxide and hydrogen sulfide in melatonin-induced tolerance of pepper (Capsicum annuum L.) plants to iron deficiency and salt stress alone or in combination

There seems to be no report in the literature on the effect of melatonin (MT) in relieving the detrimental effects of combined application of salt stress (SS) and iron deficiency (ID). Therefore, the effect of MT on the accumulation/synthesis of endogenous nitric oxide (NO) and hydrogen sulphide (H₂S) and how far these molecules are involved in MT-improved tolerance to the combined application of ID and SS in pepper (Capsicum annuum L) were tested. Hence, two individual trials were set up. The treatments in the first experiment comprised: Control, ID (0.1 mM FeSO₄), SS (100 mM NaCl) and ID + SS. The detrimental effects of combined stresses were more prominent than those by either of the single stress, with respect to growth, oxidative stress and antioxidant defense attributes. Single stress or both in combination improved the endogenous H₂S and NO, and foliar-applied MT (100 µM) led to a further increase in NO and H₂S levels. In the second experiment, 0.1 mM scavenger of NO, 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide potassium salt (cPTIO) and that of H₂S, hypotuarine (HT) were applied along with MT to get further evidence whether NO and H₂S are involved in MT-induced tolerance to ID and SS. MT combined with cPTIO and HT under a single or combined stress showed that NO effect was reversed by the NO scavenger, cPTIO, alone but the H₂S effect was inhibited by both scavengers. These findings suggested that tolerance to ID and SS induced by MT may be involved in downstream signal crosstalk between NO and H₂S.     

Differential response of cultivated rice to pathogen challenge and abiotic stresses with reference to cationic peroxidase

Seedlings of cultivated rice variety ADT43 was investigated after challenging with two different abiotic (drought and salinity) and biotic (sheath blight and bacterial leaf blight pathogens) stresses. Salinity and drought stress reduced the growth of seedlings, mainly the higher conditions (100 mM NaCl and 10?days of drought, respectively). Increased level of MDA content was observed in biotic and abiotic-stress treated seedlings. The highest H₂O₂ content was observed under salinity-stressed seedlings and lower level observed under biotic stress. Superoxide dismutase activity showed a gradual decrease in all stress conditions compared to control. Salinity stress resulted in highest activity of catalase compared to biotic stress. The peroxidase activity of the seedlings was found to be increased under salt and drought stress conditions and the activity decreased under biotic stress. Drought stress resulted in induced expression of POC1 gene whereas the biotic stress showed lower expression level. Suppression of the rice peroxidase would have been the mechanism of overcoming the intrinsic defence in rice by these pathogens.     

Antioxidant responses of wheat plants under stress

Currently, food security depends on the increased production of cereals such as wheat (Triticum aestivum L.), which is an important source of calories and protein for humans. However, cells of the crop have suffered from the accumulation of reactive oxygen species (ROS), which can cause severe oxidative damage to the plants, due to environmental stresses. ROS are toxic molecules found in various subcellular compartments. The equilibrium between the production and detoxification of ROS is sustained by enzymatic and nonenzymatic antioxidants. In the present review, we offer a brief summary of antioxidant defense and hydrogen peroxide (H₂O₂) signaling in wheat plants. Wheat plants increase antioxidant defense mechanisms under abiotic stresses, such as drought, cold, heat, salinity and UV-B radiation, to alleviate oxidative damage. Moreover, H₂O₂ signaling is an important factor contributing to stress tolerance in cereals.     

Protective mechanisms of hydrogen sulfide in myocardial ischemia

Hydrogen sulfide (H₂S), which has been identified as the third gaseous signaling molecule after nitric oxide (NO) and carbon monoxide (CO), plays an important role in maintaining homeostasis in the cardiovascular system. Endogenous H₂S is produced mainly by three endogenous enzymes: cystathionine β-synthase, cystathionine γ-lyase, and 3-mercaptopyruvate sulfur transferase. Numerous studies have shown that H₂S has a significant protective role in myocardial ischemia. The mechanisms by which H₂S affords cardioprotection include the antifibrotic and antiapoptotic effects, regulation of ion channels, protection of mitochondria, reduction of oxidative stress and inflammatory response, regulation of microRNA expression, and promotion of angiogenesis. Amplification of NO- and CO-mediated signaling through crosstalk between H₂S, NO, and CO may also contribute to the cardioprotective effect. Exogenous H₂S donors are expected to become effective drugs for the treatment of cardiovascular diseases. This review article focuses on the protective mechanisms and potential therapeutic applications of H₂S in myocardial ischemia.     

H2S contributed from CSE during cellular senescence suppresses inflammation and nitrosative stress

Aging involves the time-dependent deterioration of physiological functions attributed to various intracellular and extracellular factors. Cellular senescence is akin to aging and involves alteration in redox homeostasis. This is primarily marked by increased reactive oxygen/nitrogen species (ROS/RNS), inflammatory gene expression, and senescence-associated beta-galactosidase activity, all hallmarks of aging. It is proposed that gasotransmitters which include hydrogen sulfide (H₂S), carbon monoxide (CO), and nitric oxide (NO), may affect redox homeostasis during senescence. H₂S has been independently shown to induce DNA damage and suppress oxidative stress. While an increase in NO levels during aging is well established, the role of H₂S has remained controversial. To understand the role of H₂S during aging, we evaluated H₂S homeostasis in non-senescent and senescent cells, using a combination of direct measurements with a fluorescent reporter dye (WSP-5) and protein sulfhydration analysis. The free intracellular H₂S and total protein sulfhydration levels are high during senescence, concomitant to cystathionine gamma-lyase (CSE) expression induction. Using lentiviral shRNA-mediated expression knockdown, we identified that H₂S contributed by CSE alters global gene expression, which regulates key inflammatory processes during cellular senescence. We propose that H₂S decreases inflammation during cellular senescence by reducing phosphorylation of IκBα and the p65 subunit of nuclear factor kappa B (NF-κB). H₂S was also found to reduce NO levels, a significant source of nitrosative stress during cellular senescence. Overall, we establish H₂S as a key gasotransmitter molecule that regulates inflammatory phenotype and nitrosative stress during cellular senescence.