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Currently, the field of postharvest technology is a growing research area of particular interest since the increase in world population and the need to satisfy its nutritional requirements. These aspects establish a demand to produce fruits with high market quality while minimizing the losses from production to consumption stages. In the past few years, nitric oxide (NO) has emerged as a novel gasotransmitter to improve fruit postharvest shelf-life, owing to its influence on physiological processes (including fruit ripening) and on acclimation responses to stress conditions. In this review, we summarize some of the research related to the effects of NO exposure on different fruits with the aim to extend postharvest shelf-life and quality. The protection against chilling injury and postharvest diseases are addressed. The mechanisms of NO action and its interactions with other phytohormones are analyzed. Finally, the potential use of NO donors (and other strategies destined to enhance its levels) with a technological scope is also discussed., Instituto de Fisiología Vegetal

Tomato fruit ripening and postharvest display a complex net of metabolic pathways, which are controlled by different signaling molecules and hormones that coordinate this physiological process. The inhibition of the ethylene action with 1-methylcyclopropene (1-MCP) has proved to be an excellent tool for maintaining fruit quality during postharvest. Along with these findings, nitric oxide (NO) demonstrated different effects on several species in fruit postharvest. The aim of this work was to evaluate the joint effect of 1-MCP and a NO donor (S-nitrosoglutathione, GSNO) during tomato postharvest at 23 °C. Breaker tomatoes were harvested from a local grower and divided in the following treatments: untreated, 1 mM GSNO, 0.5 μL L−1 of 1-MCP, and the combination of 1 mM GSNO + 0.5 μL L−1 1-MCP. Fruit was stored at 23 °C and analyzed after 5 and 10 d. According to the results, the combination of 1-MCP and the NO donor delayed fruit softening at days 5 and 10, reduced the ethylene synthesis significantly at day 5 and elevated the respiration rate at 10 d, compared to the individual treatments of 1-MCP and GSNO alone. However, the solely application of 1-MCP, independently from the addition of GSNO, showed a strong effect in the rise of the total titratable acidity content, together with the citric acid content. 1-MCP also up-regulated the CAT, SOD and POD activities at 5 and 10 d after harvest. These findings may suggest that some fruit quality parameters in tomato postharvest are controlled by ethylene, while others need the coordination of nitric oxide as a signal molecule.

Phosphate (P) is characterized by its low availability and restricted mobility in soils, and also by a high redistribution capacity inside plants. In order to maintain P homeostasis in nutrient restricted conditions, plants have developed mechanisms which enable P acquisition from the soil solution, and an efficient reutilization of P already present in plant cells. Nitric oxide (NO) is a bioactive molecule with a plethora of functions in plants. Its endogenous synthesis depends on internal and environmental factors, and is closely tied with nitrogen (N) metabolism. Furthermore, there is evidence demonstrating that N supply affects P homeostasis and that P deficiency impacts on N assimilation. This review will provide an overview on how NO levels in planta are affected by P deficiency, the interrelationship with N metabolism, and a summary of the current understanding about the influence of this reactive N species over the processes triggered by P starvation, which could modify P use efficiency., Instituto de Fisiología Vegetal

Blueberries are a perishable fruit that loses its firmness and weight rapidly after harvest. High concentrations of ascorbic acid (AA) and phenolic compounds such as anthocyanins are important attributes contributing to the quality of the fruit. The aim of this work is to extend the postharvest life of three blueberries cultivars; ‘Mistý and ‘Blue Cuinex́, firm and soft cultivars respectively and ‘Blue Chiṕ with intermediate firmness. The fruit was treated separately or in combination with a nitric oxide donor, S-nitrosoglutathione (GSNO) and 1-methylcyclopropene (1-MCP) and stored at 4 °C for 14 d. The best treatment for ‘Blue Cuinex́ was the combination of 1- MCP and GSNO, while ‘Mistý softened slower when treated with 1-MCP, and ‘Blue Chiṕ was not affected by treatment. The 1-MCP+ GSNO treatment maintained higher concentrations of AA and GSH in ‘Blue Cuinex́. AA concentrations declined in ‘Mistý but did not respond to GSNO. The combination of 1-MCP and GSNO extended the postharvest life of ‘Blue Cuinex́ by improving the concentrations of AA and glutathione in the berries but not for ‘Mistý. These results demonstrate that blueberry cultivars respond differentially to 1-MCP and nitric oxide, and that the sequence of application could be used to slow fruit softening and maintain antioxidant properties., Instituto de Fisiología Vegetal

Nitric oxide (NO) and potassium (K+) exert a profound influence on the acclimation of plants to multiple stress conditions. A recent report indicated that exogenous addition of an NO donor causes, under conditions of adequate K+ supply, a detrimental effect on K+ status. It remains unknown whether an exogenous NO source could negatively affect the potential capture of this element when plants are faced with a K+ shortage. In this work we offer evidence that, under conditions of K+-deprivation, the addition of the naturally occurring NO donor, S-nitrosoglutathione (GSNO), diminishes the potential inward transport of the K+-analogue rubidium (Rb+) from diluted Rb+ concentrations in Arabidopsis thaliana. Studies with the akt1-2 mutant, lacking the AKT1 inward-rectifier K+-channel involved in K+-uptake, unveiled that the effect of GSNO on Rb+-influx involves a non-AKT1 component. In addition, exposure to the NO-donor led to down-regulation of transcripts coding for the AtHAK5 K+-transporter, a major component of the K+-transport machinery in K+-deprived plants. Moreover, studies with the hak5 mutant showed that GSNO could either stimulate Rb+-uptake or does not lead to a significant effect on Rb+-uptake relative to -K+ and to -K+ in the presence of decayed GSNO, respectively, thus indicating that the presence of AtHAK5 is required for GSNO exerting an inhibitory effect.

Phosphorus (P) is a macronutrient with structural and regulatory functions, essential for energy transfer. Under limited P availability, plant cells respond to internal signals, adjusting their metabolic pathways in order to reorganize physiological priorities. This work was conducted with the aim to explore the initial changes following P deprivation, before neither growth nor photosynthesis was strongly affected. The first unifoliate leaves of nine-day-old soybean plants (Glycine max, cv. Williams 82) were analyzed. Plants grown hydroponically under control conditions (+P, nutrient solution with 500 μM H2PO4 −) were compared with those restricted in phosphate (-P, without H2PO4 − in the nutrient solution). No visible symptoms of P-deficiency, such as changes in pigment intensity and leaf number or size, were observed up to 48 h of P deprivation. Neither fresh weight nor shoot/root FW ratio was affected in plants as a consequence of early P-restriction. Shotgun proteomic analyses of leaves from plants exposed to 24 h of P-deprivation, revealed that a total of 202 proteins were exclusively detected and 232 increased their relative abundance under –P conditions. The proteins affected belong mainly to the catalytic activity group according to Gene Ontology Consortium including proteins like 6-phosphogluconate dehydrogenase (from pentose phosphate pathway), pyrophosphate-fructose 6-phosphate 1-phosphotransferase subunit alpha, alpha-1,4 glucan phosphorylases (from carbohydrate metabolic processes), and other enzymes involved in metabolic processes such as glyceraldehyde-3-phosphate dehydrogenase, pyruvate kinase and fructose-bisphosphate aldolase (from gycolisis), and nitrate reductase (from nitrogen assimilation). Early events in leaves also involve higher levels of the bioactive molecule nitric oxide (NO), detected employing confocal laser microscopy, accompanied with a parallel increase in nitrate reductase activity. Furthermore, the presence of nitrated proteins was detected using shotgun liquid chromatography–mass spectrometry (LCeMS/MS). This post-translational protein modification, affected proteins mainly related to photosynthesis (chlorophyll a–b binding protein, ribulose bisphosphate carboxylase), and some other metabolic processes (glutamine synthetase, methionine synthase, lipoxygenase). This methodology allowed the confirmation of nitration sites in proteins previously described as putatively nitrated. Taken together the data presented suggest important metabolic changes, including nitric oxide metabolism, during the first hours of P deprivation. Fil: Ramos Artuso, Facundo Antonio. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Fisiología Vegetal. Universidad Nacional de La Plata. Facultad de Ciencias Naturales y Museo. Instituto de Fisiología Vegetal; Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Agrarias y Forestales; Argentina Fil: Galatro, Andrea Verónica. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Fisiología Vegetal. Universidad Nacional de La Plata. Facultad de Ciencias Naturales y Museo. Instituto de Fisiología Vegetal; Argentina Fil: Lima, Analia Ethel. Instituto Pasteur de Montevideo; Uruguay. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; Argentina Fil: Batthyány, Carlos. Instituto Pasteur de Montevideo; Uruguay Fil: Simontacchi, Marcela Silvia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Fisiología Vegetal. Universidad Nacional de La Plata. Facultad de Ciencias Naturales y Museo. Instituto de Fisiología Vegetal; Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Agrarias y Forestales; Argentina

Plants under conditions of essential mineral deficiency trigger signaling mechanisms that involve common components. Among these components, nitric oxide (NO) has been identified as a key participant in responses to changes in nutrient availability. Usually, nutrient imbalances affect the levels of NO in specific plant tissues, via modification of its rate of synthesis or degradation. Changes in the level of NO affect plant morphology and/or trigger responses associated with nutrient homeostasis, mediated by its interaction with reactive oxygen species, phytohormones, and through post-translational modification of proteins. NO-related events constitute an exciting field of research to understand how plants adapt and respond to conditions of nutrient shortage. This review summarizes the current knowledge on NO as a component of the multiple processes related to plant performance under conditions of deficiency in mineral nutrients, focusing on macronutrients such as nitrogen, phosphate, potassium, and magnesium, as well as micronutrients such as iron and zinc. Fil: Buet, Agustina. Universidad Nacional de La Plata. Facultad de Ciencias Agrarias y Forestales; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Fisiología Vegetal. Universidad Nacional de La Plata. Facultad de Ciencias Naturales y Museo. Instituto de Fisiología Vegetal; Argentina Fil: Galatro, Andrea Verónica. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Fisiología Vegetal. Universidad Nacional de La Plata. Facultad de Ciencias Naturales y Museo. Instituto de Fisiología Vegetal; Argentina Fil: Ramos Artuso, Facundo Antonio. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Fisiología Vegetal. Universidad Nacional de La Plata. Facultad de Ciencias Naturales y Museo. Instituto de Fisiología Vegetal; Argentina Fil: Simontacchi, Marcela Silvia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Fisiología Vegetal. Universidad Nacional de La Plata. Facultad de Ciencias Naturales y Museo. Instituto de Fisiología Vegetal; Argentina

Like all living organisms, plants demand iron (Fe) for important biochemical and metabolic processes. Internal imbalances, as a consequence of insufficient or excess Fe in the environment, lead to growth restriction and affect crop yield. Knowledge of signals and factors affecting each step in Fe uptake from the soil and distribution (long-distance transport, remobilization from old to young leaves, and storage in seeds) is necessary to improve our understanding of plant mineral nutrition. In this context, the role of nitric oxide (NO) is discussed as a key player in maintaining Fe homeostasis through its cross talk with hormones, ferritin, and frataxin and the ability to form nitrosyl-iron complexes.

Improving phosphorus (P) acquisition and utilization in crops is of great importance in order to achieve a good plant nutritional state and maximize biomass production while minimizing the addition of fertilizers, and the concomitant risk of eutrophication. This study explores to which extent key processes involved in P-acquisition, and other acclimation mechanisms to low P supply in maize (Zea mays L.) plants, are affected by the addition of a nitric oxide (NO) donor (S-nitrosoglutathione, GSNO). Plants grown in a complete culture solution were exposed to four treatments performed by the combination of two P levels (0 and 0.5 mM), and two GSNO levels (0 and 0.1 mM), and responses to P-deprivation were then studied. Major plant responses related to P-deprivation were affected by the presence of the NO donor. In roots, the activity of acid phosphatases was significantly increased in P-depleted plants simultaneously exposed to GSNO. Acidification of the culture solution also increased in plants that had been grown in the presence of the NO donor. Furthermore, the potential capability displayed by roots of P-deprived plants for P-uptake, was higher in the plants that had been treated with GSNO. These results indicate that exogenous NO addition affects fundamental acclimation responses of maize plants to P scarcity, particularly and positively those that help plants to sustain P-acquisition under low P availability., Instituto de Fisiología Vegetal

The ability of plants to respond to a wide range of environmental stresses is highly flexible and finely balanced through the interaction of hormonal plant growth regulators and the redox signalling hub, which integrates information from the environment and cellular metabolism/physiology. Plant hormones produce reactive oxygen species (ROS) as second messengers in signalling cascades that convey information concerning changes in hormone concentrations and/or sensitivity to mediate a whole range of adaptive responses. Cellular redox buffering capacity that is determined largely by the abundance of ascorbate has a profound influence on the threshold at which hormone signalling is triggered and on the interactions between different hormones. Other antioxidants such as glutathione, glutaredoxins and thioredoxins are also central redox regulators of hormone signalling pathways. The complex network of cross-communication between oxidants and antioxidants in the redox signalling hub and the different hormone signalling pathways maximises productivity under stress-free situations and regulates plant growth, development, reproduction, programmed cell death and survival upon exposure to stress. This interactive network confers enormous regulatory potential because it allows plants to adapt to changing and often challenging conditions, while preventing boom or bust scenarios with regard to resources, ensuring that energy is produced and utilised in a safe and efficient manner Fil: Bartoli, Carlos Guillermo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Fisiología Vegetal. Universidad Nacional de La Plata. Facultad de Ciencias Naturales y Museo. Instituto de Fisiología Vegetal; Argentina Fil: Casalongue, Claudia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Biológicas. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Biológicas; Argentina Fil: Simontacchi, Marcela Silvia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Fisiología Vegetal. Universidad Nacional de La Plata. Facultad de Ciencias Naturales y Museo. Instituto de Fisiología Vegetal; Argentina Fil: Marquez Garcia, Belen. University of Leeds; Reino Unido Fil: Foyer, Christine H.. University of Leeds; Reino Unido

The subcellular localization of NO generation in soybean cotyledons, and the relationship between NO synthesis and in vivo chloroplast performance were studied. Employing the NO probe 4-aminomethyl-2′,7′-difluorofluorescein diacetate (DAF-FM DA) and fluorescence microscopy, a strongly punctuated fluorescence was detected in mesophyll cells. The co-localization of DAF-FM and chlorophyll fluorescence, in confocal laser microscopy images, indicated the presence of NO in the chloroplasts. NO visualization was dependent on light, seedling age, and chloroplast function throughout cotyledons lifespan. The addition of herbicides with action in chloroplasts (DCMU and paraquat) dramatically reduced the quantum yield of photosystem II (ϕPSII), and lead to images with absence of punctuated green fluorescence. Moreover, electron paramagnetic resonance signals corresponding to NO-spin trap adduct observed in cotyledon homogenates decreased significantly by the treatment with herbicides, as compared to controls. Neither chloroplast function nor NO content were significantly different in cotyledons from plants growing in the presence of ammonium or nitrate as the nitrogen source. These findings suggest that chloroplasts are organelles that contribute to NO synthesis in vivo, and that their proper functionality is essential for maintaining NO levels in soybean cotyledons. Fil: Galatro, Andrea Verónica. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Programa de Radicales Libres; Argentina. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica; Argentina Fil: Puntarulo, Susana Ángela. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Programa de Radicales Libres; Argentina. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica; Argentina Fil: Guiamet, Juan José. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Fisiología Vegetal. Universidad Nacional de La Plata. Facultad de Ciencias Naturales y Museo. Instituto de Fisiología Vegetal; Argentina Fil: Simontacchi, Marcela Silvia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Fisiología Vegetal. Universidad Nacional de La Plata. Facultad de Ciencias Naturales y Museo. Instituto de Fisiología Vegetal; Argentina

Nitric oxide (NO) is a small gaseous molecule, with a free radical nature that allows it to participate in a wide spectrum of biologically important reactions. NO is an endogenous product in plants, where different biosynthetic pathways have been proposed. First known in animals as a signaling molecule in cardiovascular and nervous systems, it has turned up to be an essential component for a wide variety of hormone-regulated processes in plants. Adaptation of plants to a changing environment involves a panoply of processes, which include the control of CO2 fixation and water loss through stomatal closure, rearrangements of root architecture as well as growth restriction. The regulation of these processes requires the concerted action of several phytohormones, as well as the participation of the ubiquitous molecule NO. This review analyzes the role of NO in relation to the signaling pathways involved in stomatal movement, plant growth and senescence, in the frame of its interaction with abscisic acid, auxins, gibberellins, and ethylene., Instituto de Fisiología Vegetal

Several apoplastic enzymes have been implicated in the control of elongation growth of plant cells. Among them, peroxidases contribute to both loosening and stiffening of the cell wall. They appear to be regulated by various mechanisms, including the action of extracellular inhibitors. To obtain evidence of the role of the enzyme-inhibitor interaction during leaf development, the intercellular washing fluids from Helianthus annuus leaves of different ages were isolated using standard methods of vacuum infiltration and centrifugation. Peroxidase activities, assessed using tetramethylbenzidine as substrate, increased during leaf development, reaching a maximum value after the leaves were fully expanded. An inhibitor, chemically characterised as ascorbate, co-localised with the enzyme in the apoplast. Moreover, there was a strong negative correlation between the action of peroxidase and the micromolar concentration of ascorbate in the apoplastic fluid. The results show that in growing leaves, the in planta ascorbate concentration is able to restrain peroxidase enzyme activity. Then, at the time of growth cessation, the loss of extracellular ascorbate relieves the inhibition on this enzyme that contributes to wall fixation.

Senescence was studied in soybean (Glycine max, var ALM 4500) cotyledons from 10- and 25-day-old seedlings. Rejuvenated cotyledons, obtained by removing epicotyls from 5-day seedlings, showed a lower decrease in the content of photosynthetic pigments from day 10 to day 25, as compared to naturally senescent cotyledons. Unlike controls, rejuvenated cotyledons showed no changes in electrolyte leakage, chlorophyll a, and carotene content during the studied period. At day 10, nitric oxide (NO) content in rejuvenated cotyledons was higher than in naturally senescent cotyledons. The role of NO in senescence was assessed by exposing seedlings to sodium nitroprusside (SNP). SNP-treated cotyledons showed a constant NO content over the studied period of 4.0 0.4 nmol g1 FW, conserved photosynthetic pigment content and lower lipid radical content, as compared to naturally senescent cotyledons. These results suggest that NO can protect against senescence-dependent alterations in cotyledons, because (i) exogenous NO can partially protect naturally senescent cotyledons and (ii) rejuvenated cotyledons, which do not show the senescent phenotype, have higher endogenous levels of NO. Fil: Jasid, Sebastian Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Programa de Radicales Libres; Argentina. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica; Argentina Fil: Galatro, Andrea Verónica. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Programa de Radicales Libres; Argentina. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica; Argentina Fil: Villordo, Juan Javier. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Programa de Radicales Libres; Argentina Fil: Puntarulo, Susana Ángela. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Programa de Radicales Libres; Argentina. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica; Argentina Fil: Simontacchi, Marcela Silvia. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Programa de Radicales Libres; Argentina. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica; Argentina

Sodium nitroprusside (SNP) and diethylenetriamine NONOate (DETA NONOate), were used as the source of exogenous NO to study the effect of NO upon germination of sorghum (Sorghum bicolor (L.) Moench) seeds through its possible interaction with iron. Modulation of cellular Fe status could be an important factor for the establishment of oxidative stress and the regulation of plant physiology. Fresh and dry weights of the embryonic axes were significantly increased in the presence of 0.1 mM SNP, as compared to control. Spin trapping EPR was used to assess the NO content in axes from control seeds after 24 h of imbibition (2.4+/-0.2 nmol NO g(-1) FW) and seeds exposed to 0.01, 0.1, and 1 mM SNP (3.1+/-0.3, 4.6+/-0.2, and 6.0+/-0.9 nmol NO g(-1) FW, respectively) and 1 mM DETA NONOate (6.2+/-0.6 nmol NO g(-1) FW). Incubation of seeds with 1 mM SNP protected against oxidative damage to lipids and maintained membrane integrity. The content of the deferoxamine-Fe (III) complex significantly increased in homogenates of axes excised from seeds incubated in the presence of 1 mM SNP or 1 mM DETA NONOate as compared to the control (19+/-2 nmol Fe g(-1) FW, 15.2+/-0.5 nmol Fe g(-1) FW, and 8+/-1 nmol Fe g(-1) FW, respectively), whereas total Fe content in the axes was not affected by the NO donor exposure. Data presented here provide experimental evidence to support the hypothesis that increased availability of NO drives not only protective effects to biomacromolecules, but to increasing the Fe availability for promoting cellular development as well.

The major feature of the plant-growth-promoting bacteria Azospirillum brasilense is its ability to modify plant root architecture. In plants, nitric oxide (NO) mediates indole-3-acetic acid (IAA)-signaling pathways leading to both lateral (LR) and adventitious (AR) root formation. Here, we analyzed aerobic NO production by A. brasilense Sp245 wild type (wt) and its mutants Faj009 (IAA-attenuated) and Faj164 (periplasmic nitrate reductase negative), and its correlation with tomato root-growth-promoting effects. The wt and Faj009 strains produced 120 nmol NO per gram of bacteria in aerated nitrate-containing medium. In contrast, Faj164 produced 5.6 nmol NO per gram of bacteria, indicating that aerobic denitrification could be considered an important source of NO. Inoculation of tomato (Solanum lycopersicum Mill.) seedlings with both wt and Faj009 induced LR and AR development. In contrast, Faj164 mutant was not able to promote LR or AR when seedlings grew in nitrate. When NO was removed with the NO scavenger 2-(4-carboxyphenyl)-4,4,5,5,-tetramethylimidazoline-1-oxyl-3-oxide (cPTIO), both LR and AR formation were inhibited, providing evidence that NO mediated Azospirillum-induced root branching. These results show that aerobic NO synthesis in A. brasilense could be achieved by different pathways and give evidence for an NO-dependent promoting activity on tomato root branching regardless of bacterial capacity for IAA synthesis.

Nitric oxide in plants may originate endogenously or come from surrounding atmosphere and soil. Interestingly, this gaseous free radical is far from having a constant level and varies greatly among tissues depending on a given plant’s ontogeny and environmental fluctuations. Proper plant growth, vegetative development, and reproduction require the integration of plant hormonal activity with the antioxidant network, as well as the maintenance of concentration of reactive oxygen and nitrogen species within a narrow range. Plants are frequently faced with abiotic stress conditions such as low nutrient availability, salinity, drought, high ultraviolet (UV) radiation and extreme temperatures, which can influence developmental processes and lead to growth restriction making adaptive responses the plant’s priority. The ability of plants to respond and survive under environmental-stress conditions involves sensing and signaling events where nitric oxide becomes a critical component mediating hormonal actions, interacting with reactive oxygen species, and modulating gene expression and protein activity. This review focuses on the current knowledge of the role of nitric oxide in adaptive plant responses to some specific abiotic stress conditions, particularly low mineral nutrient supply, drought, salinity and high UV-B radiation., Instituto de Fisiología Vegetal

Like all living organisms, plants demand iron (Fe) for important biochemical and metabolic processes. Internal imbalances, as a consequence of insufficient or excess Fe in the environment, lead to growth restriction and affect crop yield. Knowledge of signals and factors affecting each step in Fe uptake from the soil and distribution (long-distance transport, remobilization from old to young leaves, and storage in seeds) is necessary to improve our understanding of plant mineral nutrition. In this context, the role of nitric oxide (NO) is discussed as a key player in maintaining Fe homeostasis through its cross talk with hormones, ferritin, and frataxin and the ability to form nitrosyl-iron complexes.

Germination of sorghum embryonic axes, defined as the development of a root 1–2 mm long, started after 24 h of imbibition. A distinctive EPR signal for the adduct MGD–Fe–NO was detected in the homogenates from axes isolated from seeds imbibed in the presence of 12 mM nitrate in a time-dependent manner. Also a close association between MGD–Fe–NO adduct content in the homogenates and nitrate supplementation to the incubation medium was observed. The activities of the NADH-dependent nitrate reductase (NR) and nitric oxide synthase were measured in axes, by the detection of NO by EPR under conditions of maximal supplementation of substrates. A significant increase in both enzymatic activities was determined between 24 and 30 h of imbibition. Total content in sorghum axes of β-carotene and α-tocopherol increased significantly as the germination occurred, however the total content of tocopherols was not affected over the studied period. The data reported here are the first observations employing EPR to assess for NR and NOS activities simultaneously in an active growing tissue. Moreover, since the increase in NO content preceded the initiation of phase II of development and the sharp increase in oxygen consumption, a potential role for NO as a signal molecule should be considered.

The early and slight increase in oxygen uptake of the sorghum axes defines germination phase I, whereas germination phase II is characterized by the assumption of a marked rate of increase in both oxygen uptake and seed weight. The cellular non-enzymatic antioxidant capacity was determined during the early imbibition period. Over the initial 18 h of imbibition ascorbate content in sorghum seeds was not significantly affected, however, as soon as an active metabolism is established the ascorbate content in the seeds increased significantly. The same profile was observed for the total thiol content in sorghum seeds during the initial 48 h of development. Both α- and γ-tocopherol content in sorghum seeds significantly increased up to 24 h of imbibition and no further changes were detected between 24 and 48 h of incubation. The 3-desoxyanthocyanidin apigeninidin was isolated from sorghum imbibed up to 48 h and high performance liquid chromatography (HPLC) chromatograms of the isolated pigment was recorded. Apigeninidin content in sorghum seeds was significantly increased (2-fold) upon imbibition. Taken as a whole, the data presented here strongly suggest that a complex and integrated antioxidant system is triggered during the early stages of sorghum seeds development to limit oxidative-dependent deterioration at this critical period for plant survival.

The effect of addition of the nitric oxide donor S-nitrosoglutathione (GSNO) on the Zn nutritional status was evaluated in hydroponically-cultured wheat plants (Triticum aestivum cv. Chinese Spring). Addition of GSNO in Zn-deprived plants did not modify biomass accumulation but accelerated leaf senescence in a mode concomitant with accelerated decrease of Zn allocation to shoots. In well-supplied plants, Zn concentration in both roots and shoots declined due to long term exposure to GSNO. A further evaluation of net Zn uptake rate (ZnNUR) during the recovery of long-term Zn-deprivation unveiled that enhanced Zn-accumulation was partially blocked when GSNO was present in the uptake medium. This effect on uptake was mainly associated with a change of Zn translocation to shoots. Our results suggest a role for GSNO in the modulation of Zn uptake and in root-to-shoot translocation during the transition from deficient to sufficient levels of Zn-supply. Fil: Buet, Agustina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Fisiología Vegetal. Universidad Nacional de La Plata. Facultad de Ciencias Naturales y Museo. Instituto de Fisiología Vegetal; Argentina Fil: Moriconi, Jorge Ignacio. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín" (sede Chascomús). Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín" (sede Chascomús); Argentina Fil: Santa Maria, Guillermo Esteban. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín" (sede Chascomús). Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín" (sede Chascomús); Argentina Fil: Simontacchi, Marcela Silvia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Fisiología Vegetal. Universidad Nacional de La Plata. Facultad de Ciencias Naturales y Museo. Instituto de Fisiología Vegetal; Argentina

The aim of this work was to study the effect of ultraviolet-B (UV-B) exposure on oxidative status in chloroplasts isolated from soybean (Glycine max cv. Hood). Chloroplasts were isolated from soybean leaves excised from either control seedlings or those exposed to 30 and 60 kJ m -2 day -1 of UV-B radiation for 4 days. Chloroplastic oxidative conditions were assessed as carbon-centered radical, carbonyl groups and ascorbyl radical content. Treatment with UV-B increased the carbon-centered radical-dependent EPR signal significantly by 55 and 100% in chloroplasts from leaves exposed to 30 and 60 kJ m -2 day -1 UV-B, respectively, compared to radical content in chloroplasts from control leaves. The content of carbonyl groups increased by 37 and 62% in chloroplasts isolated from soybean leaves irradiated for 4 days with 30 and 60 kJ m - 2 day -1 UV-B, respectively. The content of soluble metabolites in isolated chloroplasts should not be taken as absolute in vivo values; however, these data are valuable for comparative studies. UV-B exposure did not significantly affect ascorbyl radical content compared to controls. The content of ascorbic acid and thiols in chloroplasts isolated from leaves exposed to 60 kJ m -2 day -1 UV-B was increased by 117 and 20.8%, respectively, compared to controls. Neither the content of total carotene nor that of β-carotene or α-tocopherol was affected by the irradiation. The results presented here suggest that the increased content of lipid radicals and oxidized proteins in the chloroplasts isolated from leaves exposed to UV-B could be ascribed to both the lack of antioxidant response in the lipid soluble fraction and the modest increase in the soluble antioxidant content.

Aging of petals was accompanied by a 9-fold increase in oxygen radical generation, and a 2-fold increase in the content of oxidized proteins. α-Tocopherol content increased at the onset of aging and decreased as aging progressed. Total thiol content followed the α-tocopherol pattern during aging. Petals' ascorbic acid content started to decline when the flowers were excised. Control petals were exposed to 1 mM paraquat during 24 h and neither the production of oxygen radicals nor the content of hydroperoxides were significantly affected by the treatment. Paraquat exposure did not significantly modified the activities of glutathione reductase and catalase, but superoxide dismutase activity was increased. α-Tocopherol content was not affected by paraquat, but total thiols content decreased by 50%. Flowers showing browning that involved 50–100% of the petal, were exposed to 1 mM paraquat over 24 h. The activities of superoxide dismutase, glutathione reductase, and catalase were not affected. Paraquat exposure led to a 3-fold increase in oxygen radical generation, and to a 59% increase in hydroperoxide content in aged petals. The data presented here suggest that oxidative stress and a concerted antioxidant response are associated with the initial stages of aging in Dendranthema morifolium petals.

Oxygen, although essential for the survival of aerobic organisms leads to generation of toxic species. The effect of oxygen on enzymatic and non-enzymatic antioxidants was determined to evaluate response to oxidative stress in soybean axes. Soybean seeds were incubated over nutrient solution-saturated filter paper. Different oxygen concentrations in the incubation atmosphere were maintained by gassing either N 2 (40% O 2 ), air (20% O 2 ), a commercial mix 40% O 2 + 60% N 2 (40% O 2 ) or O 2 (100% O 2 ) in closed plastic chambers. Oxidative stress was assessed by the oxidation of 2′,7′-dichlorofluorescein diacetate. The activities of antioxidant enzymes were determined spectrophotometrically. α-Tocopherol and ubiquinol-10 contents were measured by HPLC. The weight of axes was 13 ± 1 and 27 ± 3 mg/axis in the absence and presence of 20% oxygen, respectively. 2′,7′-Dichlorofluorescein diacetate oxidation was increased from 14 ± 2 to 66 ± 5 AU/min/mg FW by supplementation of 20% oxygen. Total glutathione content was 22 ± 6 and 33 ± 6 nmol/axis in axes grown in absence of oxygen and air, respectively. Ubiquinol-10 content was not affected by oxygen. α-Tocopherol content decreased from 384 ± 94 to 14 ± 3 pmol/axis in the absence or presence of 100% oxygen, respectively. The activities of antioxidant enzymes increased in axes exposed to oxygen. Our data suggest that exposure of soybean axes to oxygen leads to oxidative stress but damage by oxygen intermediates was limited by increases in the activity of both, antioxidant substances (i.e. glutathione) and antioxidant enzymes.

To characterize the physiological status of petals over the senescence period, the rate of leakage of electrolytes and weight were measured in Chrysanthemum morifolium RAM petals. Both a significant increase in electrolyte leakage to the external medium and a concomitant decrease of flower weight were observed. Lipid peroxidation was evaluated by measurements of thiobarbituric acid reactive substance (TBARS) in Chrysanthemum morifolium RAM petals during senescence, that was arbitrarily divided into five stages. TBARS content was higher in petals from stage 5 (complete wilting) than in stage 1 (blooming). Non-significant differences were detected in ethylene production between petals classified in stages 1–3 (50 ± 4 pmol/g fresh weight/h). The activity of the enzymes involved in hydroperoxide metabolism was determined. The activities of superoxide dismutase (SOD), catalase, peroxidases and ascorbate peroxidase (AP) were measured. SOD activity showed a maximum value at stage 3 (245 ± 30 U/mg protein), followed by a decline. A progressive increase in peroxidase (5-fold increase) and AP (6-fold increase) activities was measured as a function of time. Petals in stage 1 showed lower catalase activity (20.8 ± 0.5 μmol/min/mg protein) than petals in stages 2–5 (43 ± 3 μmol/min/mg protein). The data presented here suggest that lipid peroxidation and membrane damage are involved in deterioration of Chrysanthemum morifolium RAM petals. The significant increase in the activity of SOD, catalase, peroxidases and AP in the initial stages of senescence, indicates that antioxidant defenses are triggered by coordinated mechanisms to control damage by aging in petals.

SynopsisRates of leakage of protein and electrolytes were measured in soybean embryonic axes from soybean seeds stored for 8–42 months. A significant increase in the content of proteins and electrolytes was determined in the external medium where aged embryonic axes were incubated. Lipid peroxidation was evaluated by the measurement of thiobarbituric acid-reactive substances (TBARS) in isolated axes from aged seeds. No significant differences were detected either in the contentin vivonor in the rate of generationin vitro. To assessin vivooxidative condition of the axes, an assay employing 2′,7′-dichlorofluorescein (DCFH-DA) oxidation was used. Embryonic axes from seeds stored for 8 and 42 months showed 1.1±0.1 and 4.1±0.1 fluorescence units per min per axes, respectively, after 24 h of imbibition. This significant increase in fluorescence was measured at all the tested incubation times. Antioxidant defences, evaluated as chemiluminescence induced by t-butyl hydroperoxide and α-tocopherol content, showed a significant increase with storage time. These data suggest that the damage due to enhanced peroxidative events are limited by the increased amount of potent lipid soluble antioxidant.

Most of the elements involved in the integration of signals of low external K+-supply into a physiological response pathway remain essentially unknown. The aim of this work was to study the influence exerted by DELLA proteins, which are known to be key components for the control of growth, on plant responses during K+ deprivation in wheat (Triticum aestivum) by using two sets of near-isogenic lines (NILs) in the Maringa and April Bearded cultivars. After K+ shortage, the NILs of both cultivars containing the Rht-B1b,Rht-D1b alleles, which encode altered function DELLA proteins, displayed either a slight or no decrease in chlorophyll content, in contrast to the sharp decrease observed in the NILs having the wild type alleles (Rht-B1a,Rht-D1a). That difference was accompanied by a lower relative decrease of biomass accumulation only in the Maringa cultivar. In both cultivars, high chlorophyll retention was coupled with K+ starvation-induced differences in superoxide dismutase and ascorbate peroxidase activities, which were enhanced in K+-starved Rht-B1b,Rht-D1b NILs. In addition, Rht-B1b,Rht-D1b and Rht-B1a,Rht-D1a NILs markedly differed in the accumulation of the major cations Ca2+, Na+ and K+. These results suggest a major role of the Rht-1 genes in the control of physiological responses during K+ deprivation. Fil: Moriconi, Jorge Ignacio. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín" (sede Chascomús). Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín" (sede Chascomús); Argentina Fil: Buet, Agustina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Fisiología Vegetal. Universidad Nacional de La Plata. Facultad de Ciencias Naturales y Museo. Instituto de Fisiología Vegetal; Argentina Fil: Simontacchi, Marcela Silvia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Fisiología Vegetal. Universidad Nacional de La Plata. Facultad de Ciencias Naturales y Museo. Instituto de Fisiología Vegetal; Argentina Fil: Santa Maria, Guillermo Esteban. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín" (sede Chascomús). Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín" (sede Chascomús); Argentina

This work was aimed to investigate nitrosyl-Fe complexes formation by reaction of endogenous ligands and Fe, in sorghum embryonic axes exposed to NO-donors. Electron paramagnetic resonance (EPR) was employed to detect the presence of nitrosyl-Fe complexes in plant embryos, as well as changes in labile iron pool (LIP). Nitrosyl-Fe complexes formation was detected in sorghum embryonic axes homogenates incubated in vitro in the presence of 1 mM of NO donors: diethylenetriamine NONOate (DETA NONOate), S-nitrosoglutathione (GSNO) and sodium nitroprusside (SNP). In axes isolated from seeds incubated in vivo in the presence of 1 mM SNP for 24 h, the content of NO was increased by 2-fold, and the EPR spectrum from mononitrosyl-Fe complexes (MNIC) was observed with a concomitant increase in the fresh weight of sorghum axes. The simultaneous exposure to deferoxamine and the NO donor precluded the increase in fresh weight observed in the presence of excess NO. While total Fe content in the axes isolated from seeds exposed to 1 mM SNP was not significantly affected as compared to control axes, the LIP was increased by over 2-fold.The data reported suggest a critical role for the generation of complexes between Fe and NO when cells faced a situation leading to a significant increase in NO content. Moreover, demonstrate the presence of MNICs as one of the important components of the LIP, which could actively participate in Fe cellular mobilization. © 2011 Elsevier Ireland Ltd. All rights reserved.

Iron is a key element in plant nutrition. Iron deficiency as well as iron overload results in serious metabolic disorders that affect photosynthesis, respiration and general plant fitness with direct consequences on crop production. More than 25% of the cultivable land possesses low iron availability due to high pH (calcareous soils). Plant biologists are challenged by this concern and aimed to find new avenues to ameliorate plant responses and keep iron homeostasis under control even at wide range of iron availability in various soils. For this purpose, detailed knowledge of iron uptake, transport, storage and interactions with cellular compounds will help to construct a more complete picture of its role as essential nutrient. In this review, we summarize and describe the recent findings involving four central players involved in keeping cellular iron homeostasis in plants: nitric oxide, ferritin, frataxin and nitrosyl iron complexes. We attempt to highlight the interactions among these actors in different scenarios occurring under iron deficiency or iron overload, and discuss their counteracting and/or coordinating actions leading to the control of iron homeostasis. Fil: Ramírez, Leonor. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Mar del Plata. Instituto de Investigaciones Biológicas; Argentina. Universidad Nacional de Mar del Plata; Argentina Fil: Simontacchi, Marcela Silvia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico La Plata. Instituto de Fisiología Vegetal; Argentina. Universidad Nacional de La Plata; Argentina Fil: Murgia, Irene. Universita Degli Studi Di Milano; Italia Fil: Zabaleta, Eduardo Julian. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Mar del Plata. Instituto de Investigaciones Biológicas; Argentina. Universidad Nacional de Mar del Plata; Argentina Fil: Lamattina, Lorenzo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Mar del Plata. Instituto de Investigaciones Biológicas; Argentina. Universidad Nacional de Mar del Plata; Argentina

NO is an inorganic free radical gaseous molecule that has been shown to play an unprecedented range of roles in biological systems. Taking into account these numerous functions and the reports indicating that NO can regulate processes related to plant growth and development, endogenous sources of NO need to be clarified as well as the effect of the variations of NO levels upon the plant life cycle. We consider here the main endogenous sources of cellular NO in plant tissues, and the contribution of enzymatic sources upon seed germination. Non-enzymatic generation of NO from NO2 − under conditions of low pH could be of considerable importance since significant amounts of NO2 − can be found in plant tissues. However, at least under the reported experimental conditions, enzymatic activity seems to be more relevant to total NO generation, allowing a strict control of NO steady state concentration. The complexity of the overall scenario presented here shows the need for further studies into NO production and consumption pathways in germinating seeds exposed to fluctuating environmental conditions.

Nitric oxide (NO) generation by soybean (Glycine max, var ADM 4800) chloroplasts was studied by electron paramagnetic resonance (EPR) spin-trapping technique.1 Both nitrite and L-arginine (arg) are the required substrates for enzymatic activities considered as possible sources of NO in plants. Soybean chloroplasts showed a NO production of 3.2 +/- 0.2 nmol min(-1) mg(-1) protein in the presence of 1 mM NaNO(2). Chloroplasts incubated with 1 mM arg showed a NO production of 0.76 +/- 0.04 nmol min(-1) mg(-1) protein. This production was inhibited when chloroplasts were incubated in presence of NOS-inhibitors L-NAME and L-NNA. In vitro exposure of chloroplasts to a NO-donor (GSNO) decreased both ascorbyl radical content and the activity of ascorbate peroxidase, without modification of the total ascorbate content. Exposure of the isolated chloroplasts to a NO-donor decreased lipid radical content in membranes, however, incubation in the presence of 25 microM peroxynitrite (ONOO(-)) led to an increase in lipid-derived radicals (34%). The effect of ONOO(-) on protein oxidation was determined by western blotting, showing an increase in carbonyl content either in stroma or thylakoid proteins as compared to control. Taken as a whole, NO seems to be an endogenous metabolite in soybean chloroplasts and reactive nitrogen species could exert either antioxidant or prooxidant effects on chloroplasts, since both a decreased lipid radical content in membranes and a decrease in the activity of ascorbate peroxidase were observed after exposure to a NO donor.

Nitric oxide (NO) generation by soybean (Glycine max var. ADM 4800) chloroplasts was studied as an endogenous product assessed by the electron paramagnetic resonance spin-trapping technique. Nitrite and L-arginine (Arg) are substrates for enzymatic activities considered to be the possible sources of NO in plants. Soybean chloroplasts showed a NO production of 3.2 ± 0.2 nmol min-1 mg-1 protein in the presence of 1 mM NaNO 2. Inhibition of photosynthetic electron flow by 3-(3,4- dichlorophenyl)-1,1-dimethyl urea resulted in a lower rate (1.21 ± 0.04 nmol min-1 mg-1 protein) of NO generation. Chloroplasts incubated with 1 mM Arg showed NO production of 0.76 ± 0.04 nmol min -1 mg-1 protein that was not affected either by omission of Ca2+ or by supplementation with Ca2+ and calmodulin to the incubation medium. This production was inhibited when chloroplasts were incubated in the presence of NO synthase inhibitors Nω,-nitro- L-Arg methyl ester hydrochloride and Nω-nitro-L-Arg. In vitro exposure of chloroplasts to an NO donor (250 μM S-nitrosoglutathione) decreased lipid radical content in membranes by 29%; however, incubation in the presence of 25 μM peroxynitrite (ONOO-) led to an increase in lipid-derived radicals (34%). The effect of ONOO- on protein oxidation was determined by western blotting, showing an increase in carbonyl content either in stroma or thylakoid proteins as compared to controls. Moreover, ONOO- treatment significantly affected both O2 evolution and chlorophyll fluorescence in thylakoids. Data reported here suggest that NO is an endogenous metabolite in soybean chloroplasts and that reactive nitrogen species could exert either antioxidant or prooxidant effects on chloroplast macromolecules., Instituto de Fisiología Vegetal

The amount of iron within the cell is carefully regulated in order to provide an adequate level of micronutrient while preventing its accumulation and toxicity. Iron excess is believed to generate oxidative stress, understood as an increase in the steady-state concentration of oxygen radical intermediates. Nitric oxide (NO) is an inorganic free-radical gaseous molecule which has been shown over the last decade to play an unprecedented variety of roles in biological systems. The effect of nitrogen reactive species may explain the iron sequestration pattern that characterizes macrophages under inflammatory conditions. From a patho-physiological viewpoint, further studies are required to assess the usefulness of this mechanism to minimize formation and release of free radicals in diseased tissues. However, contrary to the deleterious effects of the reactive nitrogen oxide species formed from either NO/O2 and NO/O2−, it has been pointed out that NO shows antioxidant properties. A number of studies have described the complex relationships between iron and NO, but controversy remains as to the influence and significance of iron on inflammatory NO production. To explore the initial steps of the effects triggered by LPS administration in the presence of excess iron, male Wistar rats were treated with: lipopolysaccharide from Escherichia coli (serotype 0127:B8) (LPS); iron-dextran; or iron-dextran plus LPS and liver samples were taken after 6 h. EPR spectra of NO-Hb in the venous blood were determined at 77 K. Iron-dextran administered to rats intraperitoneally resulted predominantly in iron uptake by the liver Kupffer cells and led to an increased NO level in blood in the presence of LPS. Further studies will be required to assess the complex role of the Kupffer cells on iNOS induction and NO production.

In this report, we demonstrate that nitric oxide (NO) mediates the auxin response leading the adventitious root formation. A transient increase in NO concentration was shown to be required and to be part of the molecular events involved in adventitious root development induced by indole acetic acid

The objective of this work was to explore the hypothesis that nitric oxide (NO) affects Fe bioavailability in sorghum (Sorghum bicolor (L.) Moench) embryonic axes. NO content was assessed in embryonic axes isolated from seeds control or exposed to NO-donors, employing spin trapping electron paramagnetic resonance (EPR) methodology. NO donors such as sodium nitroprusside (SNP) and diethylenetriamine NONOate (DETA NONOate), released NO that permeated inside the axes increasing NO content. Under these conditions low temperature EPR was employed to study the labile iron pool. A 2.5 fold increase was observed in NO steady state concentration after 24 h of exposure to NO donors that was correlated to a 2 fold increase in the Fe labile pool, as compared to control axes. This observation provides experimental evidence for a potential role of NO in Fe homeostasis.

The purpose of the present work was to evaluate both oxidative stress and the antioxidant response system in leaves from wheat (Triticum aestivum cv. Buck Poncho) subjected sequentially to drought and watering. Drought was imposed by withholding water until soil water potential reached - 2.0 MPa and maintained under those conditions for 24 h. DCFDA oxidation by wheat leaves was not significantly affected by drought, but watering led to an approximately 2-fold increase in DCFDA oxidation rate. However, no significant effect either on lipid radical content or on hydroperoxide content was measured after drought and drought followed by watering. Microsomes isolated from leaves exposed to drought, and from leaves exposed to drought followed by watering, generated a significantly higher amount of hydroxyl radical as compared to microsomes isolated from control leaves, suggesting a higher production of hydroxyl radical in the cellular water-soluble phase, after drought and watering as compared to control values. The content of α-tocopherol in wheat leaves was increased 2.4-fold after drought and β-carotene content was increased by 2.6-fold after drought. Hydration lowered lipid-soluble antioxidant content to control values. Total thiol content was increased by 70% after drought, and watering did not significantly alter the enhanced values. Drought decreased by 28.5% the content of reduced ascorbic acid. Taken as a whole, active species formed at wheat membranes after exposure to moderate water stress, are efficiently removed upon rehydration by reaction with an increased content of α-tocopherol and β-carotene. Moreover, a co-ordinated response involving glutathione reductase activity, thiols and ascorbic acid is triggered to limit free radical dependent effects., Instituto de Fisiología Vegetal

The objective of this work was to study the role of free radicals in relation to ethylene production during ageing of cut carnation petals. Ethylene production by freshly cut flowers was negligible, but 8 d after cutting ethylene production began to increase and reached a peak by day 9, before beginning to decline again. The efflux of electrolytes (membrane damage index) increased 101% and 2, 7-dichlorofluorescein oxidation rate (oxidative stress index) increased 53% from day 8 to day 11 after detachment. Ethylene peak was either (a) not affected significantly by the supplementation of exogenous ethylene on the day of cutting, or (b) expanded between days 7-9 after ethylene supplementation on day 6 of cutting, or (c) was inhibited by amino-oxyacetic acid and paraquat treatments. After ethylene supplementation, conductivity and 2,7-dichlorofluorescein oxidation increased significantly as compared to control petals, and the activity of antioxidant enzymes was not affected. However, both w-tocopherol and glutathione content decreased significantly after ethylene supplementation on day 6 after detachment. Amino-oxyacetic acid treatment prevented the increases in conductivity and 2,7dichlorofluorescein oxidation, did not alter the activities of antioxidant enzymes and significantly increased the content of a-tocopherol and glutathione as compared to control carnation petals. Paraquat treatment paralleled qualitatively ethylene supplementation after 6 d of cutting. Taken as a whole, the data presented here may be understood as experimental evidence of a close association between ethylene production and oxidative stress in ageing of cut carnations., Instituto de Fisiología Vegetal

The content of [alpha]-tocopherol ([alpha]T) in isolated soybean (Glycine max, var Hood) embryonic axes was measured upon germination. Dry, high-vigor axes contained 1.2 [plus or minus] 0.1, nmol/axis and after an increase during the initial 6 h of imbibition, there was a decline to 1.0 [plus or minus] 0.1 nmol/axis at 24 h of incubation. Incubation in the presence of the redox-cycling agent paraquat (4 mM) for 24 h increased the [alpha]T content to 1.9 [plus or minus] 0.2 nmol/axis. When the incubation medium was supplemented with 500 [mu]M Fe-EDTA over 24 h, the content of [alpha]T increased to 1.8 [plus or minus] 0.1 nmol/axis. Isolated axes from soybean seeds stored for 56 months contained 6.5 [plus or minus] 0.3 nmol of [alpha]T/axis after 24 h of imbibition as compared to 1.0 [plus or minus] 0.1 nmol of [alpha]T/axis in axes from soybean seeds stored for 8 months. In all of these experimental situations, oxidant production as assessed in vivo by a fluorometric assay was increased by 4 mM paraquat (8-fold), 500 [mu]M iron (2-fold), and 56 months of storage (4-fold) after 24 h of imbibition. The data presented here suggest that the cellular content of [alpha]T is physiologically adjusted as a response to conditions of oxidative stress.

The generation of active oxygen species by microsomes isolated from soybean seedlings was studied. NADPH-dependent superoxide anion production was 5.0 +/- 0.4 nmol . min(-1) mg(-1) of microsomal protein. Hydrogen peroxide generation by microsomes was 1.40 +/- 0.05 nmol . min(-1) mg(-1) of protein. Hydroxyl radical production, in the presence of ferric EDTA, evaluated through the generation of formaldehyde from dimethyl sulfoxide or tert-butyl alcohol was 0.50 +/- 0.04 and 0.44 +/- 0.03 nmol . min(-1) mg(-1), respectively. NADH proved to be suitable as cofactor for oxygen radical generation by microsomes from soybean seedlings. Because transition metals are implicated in radical generation by biological systems, the ability of microsomal membranes to reduce iron complexes was studied. Ferric ATP, ferric citrate, ferric ADP, ferric diethylenetriamine pentaacetic acid, and ferric EDTA were efficiently reduced in the presence of either NADPH or NADH as cofactor. The pattern of effectiveness of the different ferric complexes, on superoxide anion, hydrogen peroxide, and hydroxyl radical production, was similar to that found with animal microsomes. The data presented here indicate that microsomal ability to catalyze oxygen radical generation must be considered as an important contribution to cellular radical steady-state concentrations in cells from soybean seedlings.