Melatonin mediated tolerance to benzalkonium chloride phytotoxicity through improved growth, photochemical reactions, and antioxidant system in wild-type and snat2 mutant Arabidopsis lines.

Melatonin (Mel) is a phytohormone that plays a crucial role in various plant processes, including stress response. Despite numerous studies on the role of Mel in stress resistance, its significance in plants exposed to benzalkonium chloride (BAC) pollution remains unexplored. BAC, a common antiseptic, poses a threat to terrestrial plants due to its widespread use and inefficient removal, leading to elevated concentrations in the environment. This study investigated the impact of BAC (0.5 mg L-1) pollution on wild-type Col-0 and snat2 knockout mutant Arabidopsis lines, revealing reduced growth, altered water relations, and gas exchange parameters. On the other hand, exogenous Mel (100 µM) treatments mitigated BAC-induced phytotoxicity and increased the growth rate by 1.8-fold in Col-0 and 2-fold in snat2 plants. snat2 mutant seedlings had a suppressed carbon assimilation rate (A) under normal conditions, but BAC contamination led to further A repression by 71% and 48% in Col-0 and snat2 leaves, respectively. However, Mel treatment on stressed plants was successful in improving Fv/Fm and increased the total photosynthesis efficiency by regulating photochemical reactions. Excessive H2O2 accumulation in the guard cells of plants exposed to BAC pollution was detected by confocal microscopy. Mel treatments triggered almost all antioxidant enzyme activities (except POX) in both Arabidopsis lines under stress. This enhanced antioxidant activity, facilitated by foliar Mel application, contributed to the alleviation of oxidative damage, regulation of photosynthesis reactions, and promotion of plant growth in Arabidopsis. In addition to corroborating results observed in many agricultural plants regarding the development of tolerance to environmental stresses, this study provides novel insights into the action mechanisms of Mel under the emerging pollutant benzalkonium chloride.

Alternative electron sinks in chloroplasts and mitochondria of halophytes as a safety valve for controlling ROS production during salinity.

Electron flow through the electron transport chain (ETC) is essential for oxidative phosphorylation in mitochondria and photosynthesis in chloroplasts. Electron fluxes depend on environmental parameters, e.g., ionic and osmotic conditions and endogenous factors, and this may cause severe imbalances. Plants have evolved alternative sinks to balance the reductive load on the electron transport chains in order to avoid overreduction, generation of reactive oxygen species (ROS), and to cope with environmental stresses. These sinks act primarily as valves for electron drainage and secondarily as regulators of tolerance-related metabolism, utilizing the excess reductive energy. High salinity is an environmental stressor that stimulates the generation of ROS and oxidative stress, which affects growth and development by disrupting the redox homeostasis of plants. While glycophytic plants are sensitive to high salinity, halophytic plants tolerate, grow, and reproduce at high salinity. Various studies have examined the ETC systems of glycophytic plants, however, information about the state and regulation of ETCs in halophytes under non-saline and saline conditions is scarce. This review focuses on alternative electron sinks in chloroplasts and mitochondria of halophytic plants. In cases where information on halophytes is lacking, we examined the available knowledge on the relationship between alternative sinks and gradual salinity resilience of glycophytes. To this end, transcriptional responses of involved components of photosynthetic and respiratory ETCs were compared between the glycophyte Arabidopsis thaliana and the halophyte Schrenkiella parvula, and the time-courses of these transcripts were examined in A. thaliana. The observed regulatory patterns are discussed in the context of reactive molecular species formation in halophytes and glycophytes.

Role of Abscisic Acid, Reactive Oxygen Species, and Ca2+ Signaling in Hydrotropism-Drought Avoidance-Associated Response of Roots.

Plant roots exert hydrotropism in response to moisture gradients to avoid drought stress. The regulatory mechanism underlying hydrotropism involves novel regulators such as MIZ1 and GNOM/MIZ2 as well as abscisic acid (ABA), reactive oxygen species (ROS), and Ca2+ signaling. ABA, ROS, and Ca2+ signaling are also involved in plant responses to drought stress. Although the mechanism of moisture gradient perception remains largely unknown, the sensory apparatus has been reported to reside in the root elongation zone rather than in the root cap. In Arabidopsis roots, hydrotropism is mediated by the action of MIZ1 and ABA in the cortex of the elongation zone, the accumulation of ROS at the root curvature, and the variation in the cytosolic Ca2+ concentration in the entire root tip including the root cap and stele of the elongation zone. Moreover, root exposure to moisture gradients has been proposed to cause asymmetric ABA distribution or Ca2+ signaling, leading to the induction of the hydrotropic response. A comprehensive and detailed analysis of hydrotropism regulators and their signaling network in relation to the tissues required for their function is apparently crucial for understanding the mechanisms unique to root hydrotropism. Here, referring to studies on plant responses to drought stress, we summarize the recent findings relating to the role of ABA, ROS, and Ca2+ signaling in hydrotropism, discuss their functional sites and plausible networks, and raise some questions that need to be answered in future studies.

Roles of Reactive Carbonyl Species (RCS) in Plant Response to Abiotic Stress.

Abiotic and biotic stress conditions lead to production of reactive carbonyl species (RCS) which are lipid peroxide derivatives and have detrimental effects on plant cells especially at high concentrations. There are several molecules that can be classified in RCS; among them, 4-hydroxy-(E)-2-nonenal (HNE) and acrolein are widely recognized and studied because of their toxicity. The toxicity mechanisms of RCS are well known in animals but their roles in plant systems especially signaling aspects in metabolism need to be addressed. This chapter focuses on the production mechanisms of RCS in plants as well as how plants scavenge and modify them to prevent irreversible damage in the cell. We aimed to get a comprehensive look at the literature to summarize the signaling roles of RCS in plant metabolism and their interaction with other signaling mechanisms such as highly recognized reactive oxygen species (ROS) signaling. Changing climate promotes more severe abiotic stress effects on plants which also decrease yield on the field. The effects of abiotic stress conditions on RCS metabolism are also gathered in this chapter including their signaling roles during abiotic stresses. Different methods of measuring RCS in plants are also presented in this chapter to draw more attention to the study of RCS metabolism in plants.

Involvement of GLR-mediated nitric oxide effects on ROS metabolism in Arabidopsis plants under salt stress.

Plant glutamate receptor-like channels (GLRs) play important roles in plant development, immune response, defense signaling and Nitric oxide (NO) production. However, their involvement in abiotic stress responses, particularly in regulating Reactive Oxygen Species (ROS), is not well understood. This study aimed to investigate GLR-mediated NO production on ROS regulation in salt-stressed cells. To achieve this, Arabidopsis thaliana Columbia (Col-0) were treated with NaCl, glutamate antagonists [(DNQX (6,7-dinitroquinoxaline-2,3-dione and AP-5(D-2-amino-5-phosphono pentanoic acid)], and NO scavenger [cPTIO (2-(4-Carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide potassium salt)]. Salt-stressed plants in combination with DNQX and AP-5 have exhibited higher increase in lipid peroxidation (TBARS), hydrogen peroxide (H2O2) and superoxide radical (O-2) contents as compared to solely NaCl-treated plants. Furthermore, NO and total glutathione contents, and S-nitrosoglutathione reductase (GSNOR) activity decreased with these treatments. AP-5 and DNQX increased the activities of NADPH oxidase (NOX), catalase (CAT), peroxidase (POX), cell wall peroxidase (CWPOX) in salt-stressed Arabidopsis leaves. However, their activities (except NOX) were significantly inhibited by cPTIO. Conversely, the combination of NaCl and GLR antagonists, NO scavenger decreased the activities of ascorbate peroxidase (APX), superoxide dismutase (SOD), glutathione reductase (GR), dehydroascorbate reductase (DHAR) and monodehydroascorbate reductase (MDHAR) resulting in elevated GSSG levels, a low GSH/GSSG ratio, impaired ROS scavenging, excessive ROS accumulation and cell membrane damage. The findings of this study provide evidence that GLR-mediated NO plays a crucial role in improvement of the tolerance of Arabidopsis plants to salt-induced oxidative stress. It helps to maintain cellular redox homeostasis by reducing ROS accumulation and increasing the activity of SOD, GSNOR, and the ASC-GSH cycle enzymes.

Differential regulation of reactive oxygen species in dimorphic chloroplasts of single cell C4 plant Bienertia sinuspersici during drought and salt stress.

Single cell C4 (SCC4) plants, discovered around two decades ago, are promising materials for efforts for genetic engineering of C4 photosynthesis into C3 crops. Unlike C4 plants with Kranz anatomy, they exhibit a fully functional C4 photosynthesis in just a single cell and do not require mesophyll and bundle sheath cell spatial separation. Bienertia sinuspersici is one such SCC4 plant, with NAD-malic enzyme (NAD-ME) subtype C4 photosynthesis. Its chlorenchyma cell consist of two compartments, peripheral compartment (PC), analogous to mesophyll cell, and central compartment (CC), analogous to bundle sheath cell. Since oxidative stress creates an important constraint for plants under salinity and drought, we comparatively examined the response of enzymatic antioxidant system, H2O2 and TBARS contents, peroxiredoxin Q, NADPH thioredoxin reductase C, and plastid terminal oxidase protein levels of PC chloroplasts (PCC) and CC chloroplasts (CCC). Except for protein levels, these parameters were also examined on the whole leaf level, as well as catalase and NADPH oxidase activities, water status and growth parameters, and levels of C4 photosynthesis related transcripts. Many C4 photosynthesis related transcript levels were elevated, especially under drought. Activities of dehydroascorbate reductase and especially peroxidase were elevated under drought in both compartments (CCC and PCC). Even though decreases of antioxidant enzyme activities were more prevalent in PCC, and the examined redox regulating protein levels, especially of peroxiredoxin Q, were elevated in CCC under both stresses, PCC was less damaged by either stress. These suggest PCC is more tolerant and has other means of preventing or alleviating oxidative damage.

A stratagem for primary root elongation under moderate salt stress in the halophyte Schrenkiella parvula.

Schrenkiella parvula, an Arabidopsis-related halophyte, grows around Lake Tuz (Salt) in Turkey and can survive up to 600 mM NaCl. Here, we performed physiological studies on the roots of S. parvula and A. thaliana seedlings cultivated under a moderate salt condition (100 mM NaCl). Interestingly, S. parvula germinated and grew at 100 mM NaCl, but germination did not occur at salt concentrations above 200 mM. In addition, primary roots elongated much faster at 100 mM NaCl, while being thinner with fewer roots hair, than under NaCl-free conditions. Salt-induced root elongation was due to epidermal cell elongation, but meristem size and meristematic DNA replication were reduced. The expression of genes related to auxin response and biosynthesis was also reduced. Application of exogenous auxin abolished the changes in primary root elongation, suggesting that auxin reduction is the main trigger for root architectural changes in response to moderate salinity in S. parvula. In A. thaliana seeds, germination was maintained up to 200 mM NaCl, but post-germination root elongation was significantly inhibited. Furthermore, primary roots did not promote elongation even under fairly low salt conditions. Compared to A. thaliana, cell death and ROS content in primary roots of salt-stressed plants were significantly lower in S. parvula. These changes in the roots of S. parvula seedlings may be an adaptive strategy to reach lower salinity by advancing into deeper soils, while being impaired by moderate salt stress.

Nanomaterial sulfonated graphene oxide advances the tolerance against nitrate and ammonium toxicity by regulating chloroplastic redox balance, photochemistry of photosystems and antioxidant capacity in Triticum aestivum.

The current study was designed to assess nanomaterial sulfonated graphene oxide (SGO) potential in improving tolerance of wheat chloroplasts against nitrate (NS) and ammonium (AS) toxicity. Triticum aestivum cv. Ekiz was grown under SGOs (50-250-500 mg L-1) with/without 140 mM NS and 5 mM AS stress. SGOs were eliminated the adverse effects produced by stress on chlorophyll fluorescence, potential photochemical efficiency and physiological state of the photosynthetic apparatus. SGO reversed the negative effects on these parameters. Upon SGOs exposure, the induced expression levels of photosystems-related reaction center proteins were observed. SGOs reverted radical accumulation triggered by NS by enabling the increased superoxide dismutase (SOD) activity and ascorbate (AsA) regeneration. Under AS, the turnover of both AsA and glutathione (GSH) was maintained by 50-250 mg L-1 SGO by increasing the enzymes and non-enzymes related to AsA-GSH cycle. 500 mg L-1 SGO prevented the radical over-accumulation produced by AS via the regeneration of AsA and peroxidase (POX) activity rather than GSH regeneration. 50-250 mg L-1 SGO protected from the NS+AS-induced disruptions through the defense pathways connected with AsA-GSH cycle represented the high rates of AsA/DHA and, GSH/GSSG and GSH redox state. Our findings specified that SGO to NS and AS-stressed wheat provides a new potential tool to advance the tolerance mechanism.

Pretreatment of seeds with hydrogen peroxide improves deep-sowing tolerance of wheat seedlings.

Drought is a prevalent natural factor limiting crop production in arid regions across the world. To overcome this limitation, seeds are sown much deeper to boost germination by soil moisture produced by underground water. Seed pretreatment can effectively induce deep-sowing tolerance in plants. In the present study, we evaluated whether H2O2 pretreatment of seeds can initiate metabolic changes and lead to improved deep-sowing tolerance in wheat. Pretreatment with 0.05 µM H2O2 promoted first internode elongation by 13% in the deep-sowing tolerant wheat cultivar "Tir" and by 32% in the sensitive cultivar "Kiraç-66" under deep-sowing conditions, whereas internode elongation was inhibited by diphenyleneiodonium chloride. In contrast to Tir seedlings, H2O2 levels in the first internode of Kiraç-66 seedlings increased under deep-sowing condition in the H2O2-treated group compared to controls. Moreover, these seedlings had significantly lower catalase (CAT), peroxidase (POX), and ascorbate peroxidase (APX) activities but higher NADPH oxidase (NOX) and superoxide dismutase (SOD) activities under the same conditions, which consequently induced greater H2O2 accumulation. Contrary to Tir, both total glutathione and glutathione S-transferase (GST) activity decreased in Kiraç-66 after deep-sowing at 10 cm. However, H2O2 treatment increased the total glutathione amounts and the activities of glutathione-related enzymes (except GST and GPX) in the first internode of Kiraç-66. Taken together, these data support that H2O2 acts as a signaling molecule in the activation of antioxidant enzymes (specifically NOX, SOD, and CAT), regulation of both glutathione-related enzymes and total glutathione content, and upregulation of the cell wall-loosening protein gene TaEXPB23.

Differential responses of the scavenging systems for reactive oxygen species (ROS) and reactive carbonyl species (RCS) to UV-B irradiation in Arabidopsis thaliana and its high altitude perennial relative Arabis alpina.

The present work aimed to compare antioxidant response and lipid peroxide detoxification capacity of an arctic-alpine species Arabis alpina to its close relative model species Arabidopsis thaliana under acute short duration (3 h and 6 h) UV-B stress (4.6 and 8.2 W/m2). After 3 and 6 h exposure to UV-B, A. alpina showed lower lipid peroxidation and H2O2 accumulation when compared to A. thaliana. Moreover, Fv/Fm value of A. thaliana dropped to 0.70, while A. alpina dropped to 0.75 indicating better protection of PSII in this species. For elucidation of the antioxidant response, activities of superoxide dismutase (SOD), catalase (CAT), peroxidase (POX), ascorbate peroxidase (APX), glutathione reductase (GR) and dehydroascorbate reductase (DHAR) were measured. SOD induction with 6 h of UV-B was more prominent in A. alpina. Also, A. alpina had higher chloroplastic FeSOD activity when compared to A. thaliana. APX activity was also significantly induced in A. alpina, while its activity decreased at 3 h or did not change at 6 h in A. thaliana. A. alpina was able to maintain constant CAT activity, but drastic decreases were observed in A. thaliana at both time points. Moreover, A. alpina was able to maintain or induce aldehyde dehydrogenase (ALDH), alkenal reductases (AERs) and glutathione-S-transferases (GST) activity, while an opposite trend was observed in A. thaliana. These findings indicate that A. alpina was able to maintain/induce its antioxidant defence and lipid peroxide detoxification conferring better protection against UV-B.

Day and Night Fluctuations in GABA Biosynthesis Contribute to Drought Responses in Nicotiana tabacum L.

When synchronized with the light/dark cycle the circadian rhythm is termed a diurnal rhythm and this organizes an organism's daily life cycle in relation to the metabolic shifts during the day/night cycles. This is a complex task, particularly under stress conditions. Accurate maintenance of the diurnal rhythm becomes an issue under environmental extremes, such as drought due to the impairment of metabolism, redox balance, and structural integrity. In plants, the non-proteinogenic amino acid GABA accumulates to high levels in response to several stress factors but this is not always dependent on the activation of its biosynthesis. Here we propose a regulatory role to GABA during the diurnal rhythm in plants which is similar to its function in animals where it adjusts the circadian rhythm. Here we investigated whether GABA-biosynthesis was affected by drought stress during the diurnal cycle. For this, we took samples from leaves of N. tabacum plants subjected to PEG-mediated drought stress (-0.73 MPa) during the day and night cycle during a 24 hour period. Glutamate, GABA, and proline contents, along with GDH, GAD enzyme activities and transcript profiles were analyzed. Overall, we conclude that the oscillations in GABA biosynthesis during day and night cycle have an impact on drought stress responses which needs to be elucidated by further analysis.

Melatonin mitigates UV-B stress via regulating oxidative stress response, cellular redox and alternative electron sinks in Arabidopsis thaliana.

Melatonin plays an active role in neutralizing free radicals, especially by triggering the defense system and certain enzymes that work under stress in both mammals and plant systems. Exposure to ultraviolet (UV-B) stress can be deadly for plants since UV-B can induce production of reactive oxygen species and damage nucleic acids. In the present study, to uncover the possible alleviative role of melatonin against UV-B stress, Arabidopsis thaliana plants were treated with melatonin (10 µM) and were exposed to UV-B stress for 90 min and 180 min (46 and 92 kJ m-2 d-1). Plants treated with melatonin had lower lipid peroxidation levels and higher Fv/Fm values at both time points. UV-B stress-induced activities of superoxide dismutase (SOD), glutathione reductase (GR) and ascorbate peroxidase (APX), but no additional induction was observed in melatonin treated groups. Moreover, melatonin differentially regulated the expression of glutathione peroxidase 2 (GPX2) and GPX7 genes under UV-B stress. Melatonin treatment did not have any effect on glutathione biosynthesis and catabolism genes. However, expression of alternative oxidase 1a (AOX1a) and AOX1d were lower in UV-B + melatonin treated plants when compared to only UV-B treated plants, which indicates lower oxidative load in mitochondria.

Naringenin induces tolerance to salt/osmotic stress through the regulation of nitrogen metabolism, cellular redox and ROS scavenging capacity in bean plants.

The present study was conducted to uncover underlying possible effect mechanisms of flavonoid naringenin (Nar, 0.1-0.4 mM) in nitrogen assimilation, antioxidant response, redox status and the expression of NLP7 and DREB2A, on salt (100 mM NaCl) and osmotic-stressed (10% Polyethylene glycol, -0.54 MPa) Phaseolus vulgaris cv. Yunus 90). Nar ameliorated salt/osmotic stresses-induced growth inhibition and improved the accumulation of proline, glycine betaine and choline. In response to stress, Nar increased endogenous content of nitrate (NO3-) and nitrite (NO2-) by regulating of nitrate reductase and nitrite reductase. Stress-triggered NH4+ was eliminated with Nar through increases in glutamine synthetase and glutamate synthase. After NaCl or NaCl + PEG exposure, Nar utilized the aminating activity of glutamate dehydrogenase in the conversion of NH4+. The stress-inducible expression levels of DREB2A were increased further by Nar, which might have affected stress tolerance of bean. Nar induced effectively the relative expression of NLP7 in the presence of the combination or alone of stress. Also, the impaired redox state by stress was modulated by Nar and hydrogen peroxide (H2O2) and TBARS decreased. Nar regulated the different pathways for scavenging of H2O2 under NaCl and/or PEG treatments. When Nar + NaCl exposure, the damage was removed by superoxide dismutase (SOD), catalase (CAT), POX (only at 0.1 mM Nar + NaCl) and AsA-GSH cycle. Under osmotic stress plus Nar, the protection was manifested by activated CAT and, glutathione S-transferase and the regeneration of ascorbate. 0.1 mM Nar could protect bean plant against salt/osmotic stresses, likely by regulating nitrogen assimilation pathways, improving expression levels of genes associated with tolerance mechanisms and modulating the antioxidant capacity and AsA-GSH redox-based systems.

The involvement of gamma-aminobutyric acid shunt in the endoplasmic reticulum stress response of Arabidopsis thaliana.

The endoplasmic reticulum (ER) is the main site of secretory protein production and folding and its homeostasis under environmental stress is vital for the maintenance of the protein secretory pathway. The loss of homeostasis and accumulation of unfolded proteins in the ER is referred to as ER stress. Although, γ-aminobutyric acid (GABA) is an important regulator of stress response in plants, its roles during ER stress remains unclear. This study investigated the involvement of GABA in the ER stress response of plants. For this, changes in GABA metabolism under ER stress was analysed in Arabidopsis thaliana, then to study the response of the ER-folding machinery, plants were treated with exogenous GABA under ER stress. The antibiotic tunicamycin, which inhibits N-glycosylation was used to specifically induce ER stress. This stress up-regulated the expression of five glutamate decarboxylase (GAD) genes except GAD2 and GABA content of A. thaliana plants increased with an increasing concentration of tunicamycin (0.1 µg ml-1 and 0.25 µg ml-1). Moreover, expressions of genes involved in the conversion of GABA to succinate was also induced, while genes involved in transport across plasma and mitochondrial membrane showed no response to ER stress. The exogenous treatment of plants with 1-and 5-mM GABA increased plant performance under ER stress but 0.1 mM proved ineffective. Plants treated with GABA under ER stress had decreased expression of ER stress marker genes such as BIP1, BIP3 or CNX, but the expression of genes related to ER stress perception or ER-associated protein degradation showed no changes with respect to GABA treatments.

Flavonoid Naringenin Alleviates Short-Term Osmotic and Salinity Stresses Through Regulating Photosynthetic Machinery and Chloroplastic Antioxidant Metabolism in Phaseolus vulgaris.

The current study was conducted to demonstrate the possible roles of exogenously applied flavonoid naringenin (Nar) on the efficiency of PSII photochemistry and the responses of chloroplastic antioxidant of salt and osmotic-stressed Phaseolus vulgaris (cv. Yunus90). For this aim, plants were grown in a hydroponic culture and were treated with Nar (0.1 mM and 0.4 mM) alone or in a combination with salt (100 mM NaCl) and/or osmotic (10% Polyethylene glycol, -0.54 MPa). Both caused a reduction in water content (RWC), osmotic potential (ΨΠ), chlorophyll fluorescence (Fv/Fm), and potential photochemical efficiency (Fv/Fo). Nar reversed the changes on these parameters. The phenomenological fluxes (TRo/CS and ETo/CS) altered by stress were induced by Nar and Nar led to a notable increase in the performance index (PIABS) and the capacity of light reaction [ΦPo/(1-ΦPo)]. Besides, Nar-applied plants exhibited higher specific fluxes values [ABS/RC, ETo/RC, and ΨEo/(1-ΨEo)] and decreasing controlled dissipation of energy (DIo/CSo and DIo/RC). The transcripts levels of psbA and psbD were lowered in stress-treated bean but upregulated in Nar-treated plants after stress exposure. Nar also alleviated the changes on gas exchange parameters [carbon assimilation rate (A), stomatal conductance (gs), intercellular CO2 concentrations (Ci), transpiration rate (E), and stomatal limitation (Ls)]. By regulating the antioxidant metabolism of the isolated chloroplasts, Nar was able to control the toxic levels of hydrogen peroxide (H2O2) and TBARS (lipid peroxidation) produced by stresses. Chloroplastic superoxide dismutase (SOD) activity reduced by stresses was increased by Nar. In response to NaCl, Nar increased the activities of ascorbate peroxidase (APX), glutathione reductase (GR), monodehydroascorbate reductase (MDHAR), and dehydroascorbate reductase (DHAR), as well as peroxidase (POX). Nar protected the bean chloroplasts by minimizing disturbances caused by NaCl exposure via the ascorbate (AsA) and glutathione (GSH) redox-based systems. Under Nar plus PEG, Nar maintained the AsA regeneration by the induction of MDHAR and DHAR, but not GSH recycling by virtue of no induction in GR activity and the reduction in GSH/GSSG and GSH redox state. Based on these advances, Nar protected in bean chloroplasts by minimizing disturbances caused by NaCl or PEG exposure via the AsA or GSH redox-based systems and POX activity.

Hydrogen sulfide (H2S) and nitric oxide (NO) alleviate cobalt toxicity in wheat (Triticum aestivum L.) by modulating photosynthesis, chloroplastic redox and antioxidant capacity.

The role of hydrogen sulfide (H2S)/nitric oxide (NO) in mitigating stress-induced damages has gained interest in the past few years. However, the protective mechanism H2S and/or NO has towards the chloroplast system through the regulation of redox status and activation of antioxidant capacity in cobalt-treated wheat remain largely unanswered. Triticum aestivum L. cv. Ekiz was treated with alone/in combination of a H2S donor (sodium hydrosulfide (NaHS,600µM)), a NO donor (sodium nitroprusside (SNP,100µM)) and a NO scavenger (rutin hydrate (RTN,50µM)) to assess how the donors affect growth, water relations, redox and antioxidant capacity in chloroplasts, under cobalt (Co) concentrations of 150-300 µM. Stress decreased a number of parameters (growth, water content (RWC), osmotic potential (ΨΠ), carbon assimilation rate, stomatal conductance, intercellular CO2 concentrations, transpiration rate and the transcript levels of rubisco, which subsequently disrupt the photosynthetic capacity). However, SNP/NaHS counteracted the negative effects of stress on these aforementioned parameters and RTN application with stress/non-stress was reversed these effects. Hydrogen peroxide (H2O2) and TBARS were induced under stress in spite of activated ascorbate peroxidase (APX). SNP/NaHS under stress increased activation of superoxide dismutase (SOD), peroxidase (POX), APX, glutathione reductase (GR), monodehydroascorbate reductase (MDHAR), dehydroascorbate reductase (DHAR), ascorbate (tAsA) and glutathione (GSH). In conclusion, NaHS/SNP are involved in the regulation and modification of growth, water content, rubisco activity and up-regulation of ascorbate-glutathione cycle (AsA-GSH) in chloroplast under stress.

Opportunities and Limitations of Crop Phenotyping in Southern European Countries.

The Mediterranean climate is characterized by hot dry summers and frequent droughts. Mediterranean crops are frequently subjected to high evapotranspiration demands, soil water deficits, high temperatures, and photo-oxidative stress. These conditions will become more severe due to global warming which poses major challenges to the sustainability of the agricultural sector in Mediterranean countries. Selection of crop varieties adapted to future climatic conditions and more tolerant to extreme climatic events is urgently required. Plant phenotyping is a crucial approach to address these challenges. High-throughput plant phenotyping (HTPP) helps to monitor the performance of improved genotypes and is one of the most effective strategies to improve the sustainability of agricultural production. In spite of the remarkable progress in basic knowledge and technology of plant phenotyping, there are still several practical, financial, and political constraints to implement HTPP approaches in field and controlled conditions across the Mediterranean. The European panorama of phenotyping is heterogeneous and integration of phenotyping data across different scales and translation of "phytotron research" to the field, and from model species to crops, remain major challenges. Moreover, solutions specifically tailored to Mediterranean agriculture (e.g., crops and environmental stresses) are in high demand, as the region is vulnerable to climate change and to desertification processes. The specific phenotyping requirements of Mediterranean crops have not yet been fully identified. The high cost of HTPP infrastructures is a major limiting factor, though the limited availability of skilled personnel may also impair its implementation in Mediterranean countries. We propose that the lack of suitable phenotyping infrastructures is hindering the development of new Mediterranean agricultural varieties and will negatively affect future competitiveness of the agricultural sector. We provide an overview of the heterogeneous panorama of phenotyping within Mediterranean countries, describing the state of the art of agricultural production, breeding initiatives, and phenotyping capabilities in five countries: Italy, Greece, Portugal, Spain, and Turkey. We characterize some of the main impediments for development of plant phenotyping in those countries and identify strategies to overcome barriers and maximize the benefits of phenotyping and modeling approaches to Mediterranean agriculture and related sustainability.

Understanding the Role of the Antioxidant System and the Tetrapyrrole Cycle in Iron Deficiency Chlorosis.

Iron deficiency chlorosis (IDC) is an abiotic stress often experienced by soybean, owing to the low solubility of iron in alkaline soils. Here, soybean lines with contrasting Fe efficiencies were analyzed to test the hypothesis that the Fe efficiency trait is linked to antioxidative stress signaling via proper management of tissue Fe accumulation and transport, which in turn influences the regulation of heme and non heme containing enzymes involved in Fe uptake and ROS scavenging. Inefficient plants displayed higher oxidative stress and lower ferric reductase activity, whereas root and leaf catalase activity were nine-fold and three-fold higher, respectively. Efficient plants do not activate their antioxidant system because there is no formation of ROS under iron deficiency; while inefficient plants are not able to deal with ROS produced under iron deficiency because ascorbate peroxidase and superoxide dismutase are not activated because of the lack of iron as a cofactor, and of heme as a constituent of those enzymes. Superoxide dismutase and peroxidase isoenzymatic regulation may play a determinant role: 10 superoxide dismutase isoenzymes were observed in both cultivars, but iron superoxide dismutase activity was only detected in efficient plants; 15 peroxidase isoenzymes were observed in the roots and trifoliate leaves of efficient and inefficient cultivars and peroxidase activity levels were only increased in roots of efficient plants.

Reactive oxygen species and redox regulation in mesophyll and bundle sheath cells of C4 plants.

Redox regulation, antioxidant defence, and reactive oxygen species (ROS) signalling are critical in performing and tuning metabolic activities. However, our concepts have mostly been developed for C3 plants since Arabidopsis thaliana has been the major model for research. Efforts to convert C3 plants to C4 to increase yield (such as IRRI's C4 Rice Project) entail a better understanding of these processes in C4 plants. Various photosynthetic enzymes that take part in light reactions and carbon reactions are regulated via redox components, such as thioredoxins as redox transmitters and peroxiredoxins. Hence, understanding redox regulation in the mesophyll and bundle sheath chloroplasts of C4 plants is of paramount importance: it appears impossible to utilize efficient C4 photosynthesis without understanding its exact redox needs and the regulation mechanisms used during light reactions. In this review, we discuss current knowledge on redox regulation in C3 and C4 plants, with special emphasis on the mesophyll and bundle sheath differences that are found in C4. In these two cell types in C4 plants, linear and cyclic electron transport in the chloroplasts operate differentially when compared to C3 chloroplasts, changing the redox needs of the cell. Therefore, our focus is on photosynthetic light reactions, ROS production dynamics, antioxidant defence, and thiol-based redox regulation, with the aim of providing an overview of our current knowledge.