APEX2-based proximity proteomic analysis identifies candidate interactors for Plasmodium falciparum knob-associated histidine-rich protein in infected erythrocytes.

The interaction of Plasmodium falciparum-infected red blood cells (iRBCs) with the vascular endothelium plays a crucial role in malaria pathology and disease. KAHRP is an exported P. falciparum protein involved in iRBC remodelling, which is essential for the formation of protrusions or "knobs" on the iRBC surface. These knobs and the proteins that are concentrated within them allow the parasites to escape the immune response and host spleen clearance by mediating cytoadherence of the iRBC to the endothelial wall, but this also slows down blood circulation, leading in some cases to severe cerebral and placental complications. In this work, we have applied genetic and biochemical tools to identify proteins that interact with P. falciparum KAHRP using enhanced ascorbate peroxidase 2 (APEX2) proximity-dependent biotinylation and label-free shotgun proteomics. A total of 30 potential KAHRP-interacting candidates were identified, based on the assigned fragmented biotinylated ions. Several identified proteins have been previously reported to be part of the Maurer's clefts and knobs, where KAHRP resides. This study may contribute to a broader understanding of P. falciparum protein trafficking and knob architecture and shows for the first time the feasibility of using APEX2-proximity labelling in iRBCs.

Impairment in photosynthesis induced by CAT inhibition depends on the intensity of photorespiration and peroxisomal APX expression in rice.

We have previously shown that rice plants silenced for peroxisomal ascorbate peroxidase (OsAPX4-RNAi) display higher resilience to photosynthesis under oxidative stress and photorespiratory conditions. However, the redox mechanisms underlying that intriguing response remain unknown. Here, we tested the hypothesis that favorable effects triggered by peroxisomal APX deficiency on photosynthesis resilience under CAT inhibition are dependent on the intensity of photorespiration associated with the abundance of photosynthetic and redox proteins. Non-transformed (NT) and OsAPX4-RNAi silenced rice plants were grown under ambient (AC) or high CO2 (HC) conditions and subjected to 3-amino-1,2,4-triazole (3-AT)-mediated CAT activity inhibition. Photosynthetic measurements evidenced that OsAPX4-RNAi plants simultaneously exposed to CAT inhibition and HC lost the previously acquired advantage in photosynthesis resilience displayed under AC. Silenced plants exposed to environment photorespiration and CAT inhibition presented lower photorespiration as indicated by smaller Gly/Ser and Jo/Jc ratios and glycolate oxidase activity. Interestingly, when these silenced plants were exposed to HC and CAT-inhibition, they exhibited an inverse response compared to AC in terms of photorespiration indicators, associated with higher accumulation of proteins. Multivariate and correlation network analyses suggest that the proteomics changes induced by HC combined with CAT inhibition are substantially different between NT and OsAPX4-RNAi plants. Our results suggest that the intensity of photorespiration and peroxisomal APX-mediated redox signaling are tightly regulated under CAT inhibition induced oxidative stress, which can modulate the photosynthetic efficiency, possibly via a coordinated regulation of protein abundance and rearrangement, ultimately triggered by crosstalk involving H2O2 levels related to CAT and APX activities in peroxisomes.

Toxicity of methylparaben and its chlorinated derivatives to Allium cepa L. and Eisenia fetida Sav.

Methylparaben, chloro-methylparaben, and dichloro-methylparaben were evaluated in Allium cepa at 5, 10, 50, and 100 µg/L and in Eisenia fetida at 10 and 100 µg/L. In A. cepa roots, 100 µg/L methylparaben and 50 and 100 µg/L chlorinated methylparabens reduced cell proliferation, caused cellular changes, and reduced cell viability in meristems, which caused a reduction in root growth. Furthermore, they caused drastic inhibition of catalase, ascorbate peroxidase, and superoxide dismutase; activated guaiacol peroxidase and promoted lipid peroxidation in meristematic root cells. In earthworms, after 14 days exposure to the three compounds, there were no deaths, and catalase, ascorbate peroxidase, and superoxide dismutase were not inhibited. However, guaiacol peroxidase activity and lipid peroxidation were observed in animals exposed to dichloro-methylparaben. Soils with dichloro-methylparaben also caused the escape of earthworms. It is inferred that the recurrent contamination of soils with these methylparabens, with emphasis on chlorinated derivatives, can negatively impact different species that depend directly or indirectly on soil to survive.

Chloroplastic ascorbate peroxidases targeted to stroma or thylakoid membrane: The chicken or egg dilemma.

Ascorbate peroxidases (APXs) are heme peroxidases that remove hydrogen peroxide in different subcellular compartments with concomitant ascorbate cycling. Here, we analysed and discussed phylogenetic and molecular features of the APX family. Ancient APX originated as a soluble stromal enzyme, and early during plant evolution, acquired both chloroplast-targeting and mitochondrion-targeting sequences and an alternative splicing mechanism whereby it could be expressed as a soluble or thylakoid membrane-bound enzyme. Later, independent duplication and neofunctionalization events in some angiosperm groups resulted in individual genes encoding stromal, thylakoidal and mitochondrial isoforms. These data reaffirm the complexity of plant antioxidant defenses that allow diverse plant species to acquire new means to adapt to changing environmental conditions.

Heat stress negatively affects physiology and morphology during germination of Ormosia coarctata (Fabaceae, Papilionoideae).

Research on the morphophysiological behavior of forest seeds during germination with respect to climate change is scarce. To date, there have been no studies on the biochemical or morphological aspects of Ormosia spp. In this study, we subjected Ormosia coarctata seeds to various temperature conditions to investigate temperature-dependent impacts on morphology, reactive oxygen species (ROS) generation, antioxidant systems, and storage systems. Analyses were performed on seeds exposed to 25, 35, and 40 °C for 48, 96, and 144 h. The morphology was evaluated by radiation using a Faxitron MX-20 device. ROS production (superoxide anion and hydrogen peroxide), malonaldehyde (MDA), carbonylated proteins, antioxidant enzyme activity (superoxide dismutase [SOD], ascorbate peroxidase [APX], catalase [CAT], and peroxidase [POX]), ß-carotene, lycopene, glucose, and reserve enzyme activity (α- and ß-amylase, lipase, and protease) were analyzed by spectrophotometry. Heat stress (40 °C) decreased germination by 76.2% and 78.1% (compared to 25 and 35 °C, respectively), caused damage to the external morphology of the seed, increased the content of ROS, MDA, and carbonylated proteins, and reduced APX, CAT, and POX activity. Furthermore, heat stress decreased glucose content and α-amylase activity. These results suggest that an increase of 5 °C in temperature negatively affects germination, promotes oxidative stress, and induces deterioration in O. coarctata seeds.

The role of reactive oxygen species and nitric oxide in the formation of root cortical aerenchyma under cadmium contamination.

The present study aimed to evaluate root cortical aerenchyma formation in response to Cd-driven hydrogen peroxide (H2 O2 ) production and the role of nitric oxide (NO) in the alleviation of Cd oxidative stress in maize roots and its effects on aerenchyma development. Maize plants were subjected to continuous flooding for 30 days, and the following treatments were applied weekly: Cd(NO3 )2 at 0, 10, and 50 µM and Na2 [Fe(CN)5 NO]·2H2 O (an NO donor) at 0.5, 0.1, and 0.2 µM. The root biometrics; oxidative stress indicators H2 O2 and malondialdehyde (MDA); and activities of catalase (CAT), superoxide dismutase (SOD), and ascorbate peroxidase (APX) were analyzed. The root dry and fresh masses decreased at higher concentrations of NO and Cd. H2 O2 also decreased at higher NO concentrations; however, MDA increased only at higher Cd levels. SOD activity decreased at higher concentrations of NO, but CAT activity increased. Aerenchyma development decreased in response to NO. Consequently, NO acts as an antagonist to Cd, decreasing the concentration of H2 O2 by reducing SOD activity and increasing CAT activity. Although H2 O2 is directly linked to aerenchyma formation, increased H2 O2 concentrations are necessary for root cortical aerenchyma development.

Can light intensity modulate the physiological, anatomical, and reproductive responses of soybean plants to water deficit?

Little is known about the role of light intensity in modulating plant responses to stress due to water deficit (WD). Thus, the objective of this study was to determine the WD and contrasting irradiance effects on the physiology, anatomy, and grain yield of soybean plants. The experimental design was a randomized block in a growth chamber and a 2 × 2 factorial treatment arrangement: 90% (well-watered, WW) and 40% (WD) of soil field capacities (FC); and 750 (medium irradiance, MI) and 1500 (higher irradiance, HI) µmol (photons) m-2  s-1 irradiance. The WD caused a lower photosynthetic rate - as well as observed in the light curve and in the relative parameters, such as apparent quantum efficiency -, less investment in shoot biomass and pollen grain germination, resulting in lower grain yield. However, there was an increase in non-photochemical energy dissipation, a higher concentration of total soluble sugars, proline, and malondialdehyde. The WD + MI-soybean plants developed thicker spongy parenchyma (related to higher mesophilic conductance of CO2 ). In the WW + HI condition the palisade parenchyma was thicker, conferring maintenance of photosynthetic efficiency. In addition, there was an increase in the activity of superoxide dismutase, catalase, peroxidase, and ascorbate peroxidase antioxidant enzymes in leaves due to HI, regardless of FC. This induced higher energy expenditure, reflected in the reduction of the number of leaf and branches, leaf area, dry mass of leaves and stem in the WW + HI. Interestingly, these strategies of osmotic adjustment, photoprotection, and antioxidant defenses act together in the WD + HI.

Micronutrient fertilization enhances ROS scavenging system for alleviation of abiotic stresses in plants.

Reactive oxygen species (ROS) such as hydrogen peroxide at low concentrations act as signaling of several abiotic stresses. Overproduction of hydrogen peroxide causes the oxidation of plant cell lipid phosphate layer promoting senescence and cell death. To mitigate the effect of ROS, plants develop antioxidant defense mechanisms (superoxide dismutase, catalase, guaiacol peroxidase), ascorbate-glutathione cycle enzymes (ASA-GSH) (ascorbate peroxidase, monodehydroascorbate reductase, dehydroascorbate reductase and glutathione reductase), which have the function of removing and transforming ROS into non-toxic substances to maintain cellular homeostasis. Foliar or soil application of fertilizers containing B, Cu, Fe, Mn, Mo, Ni, Se and Zn at low concentrations has the ability to elicit and activate antioxidative enzymes, non-oxidizing metabolism, as well as sugar metabolism to mitigate damage by oxidative stress. Plants treated with micronutrients show higher tolerance to abiotic stress and better nutritional status. In this review, we summarized results indicating micronutrient actions in order to reduce ROS resulting the increase of photosynthetic capacity of plants for greater crop yield. This meta-analysis provides information on the mechanism of action of micronutrients in combating ROS, which can make plants more tolerant to several types of abiotic stress such as extreme temperatures, salinity, heavy metals and excess light.

Selenium biofortification enhances ROS scavenge system increasing yield of coffee plants.

There are conclusive evidences of selenium (Se) deficiency in Brazilian soils and foods. Brazil is the largest producer and consumer of coffee worldwide, which favors agronomic biofortification of its coffee. This study aimed to evaluate effects of foliar application of three formulations and six rates of Se on antioxidant metabolism, agronomic biofortification and yield of coffee beans. Seven Se concentrations (0, 10, 20, 40, 80, 100 and 160 mg L-1) were applied from three formulations of Se (sodium selenate, nano-Se 1500, and nano-Se 5000). Selenium application up to 40 mg L-1 increased the concentration of photosynthetic pigments such as chlorophylls, pheophytins and carotenoids in coffee leaves. Foliar application of Se ranging from 20 to 80 mg L-1 decreased lipid peroxidation and concentration of hydrogen peroxide, but increased superoxide dismutase, ascorbate peroxidase, catalase and glutathione reductase activities in coffee leaves. These results indicated that foliar Se application stimulates antioxidative metabolism to mitigate reactive oxygen species. Foliar application of 20 mg Se L-1 of sodium selenate increased coffee yield by 38%, and 160 mg Se L-1 of nano-Se 5000 increased dramatically coffee yield by 42%. Selenium concentration in grains ranged from 0.116 to 4.47 mg kg-1 (sodium selenate), 4.84 mg kg-1 (nano-Se 1500) and 5.82 mg kg-1 (nano-Se 5000). The results suggest the beneficial effect of Se on the increment of photosynthetic pigments, antioxidative metabolism, increased coffee yield and nutritional quality of grains. The recommended foliar Se application in this study can mitigate abiotic stressors such as high temperatures resulting in higher yield of coffee plants.

Selenium toxicity stress-induced phenotypical, biochemical and physiological responses in rice plants: Characterization of symptoms and plant metabolic adjustment.

Selenium (Se) at low concentration is considered benefit element to plants. The range between optimal and toxic concentration of Se is narrow and varies among plant species. This study aimed to evaluate the phenotypic, physiological and biochemical responses of four rice genotypes (BRS Esmeralda, BRSMG Relâmpago, BRS Bonança and Bico Ganga) grown hydroponically treated with sodium selenate (1.5 mM L-1). Selenium treated plants showed a dramatically decrease of soluble proteins, chlorophylls, and carotenoids concentration, resulting in the visual symptoms of toxicity characterized as leaf chlorosis and necrosis. Selenium toxicity caused a decrease on shoot and root dry weight of rice plants. Excess Se increased the oxidative stress monitored by the levels of hydrogen peroxide and lipid peroxidation. The enzymatic antioxidant system (catalase, superoxide dismutase, and ascorbate peroxidase) increased in response to Se supply. Interestingly, primary metabolism compounds such as sucrose, total sugars, nitrate, ammonia and amino acids increased in Se-treated plants. The increase in these metabolites may indicate a defense mechanism for the osmotic readjustment of rice plants to mitigate the toxicity caused by Se. However, these metabolites were not effective to minimize the damages on phenotypic traits such as leaf chlorosis and reduced shoot and root dry weight in response to excess Se. Increased sugars profile combined with antioxidant enzymes activities can be an effective biomarkers to indicate stress induced by Se in rice plants. This study shows the physiological attributes that must be taken into account for success in the sustainable cultivation of rice in environments containing excess Se.

Selenate and selenite affect photosynthetic pigments and ROS scavenging through distinct mechanisms in cowpea (Vigna unguiculata (L.) walp) plants.

Selenium (Se) is a beneficial element to higher plants. Application of Se at low concentrations enhances the antioxidant metabolism reducing the reactive oxygen species (ROS) generated by plant membrane cells. This study aimed to evaluate how the application of Se in the forms sodium selenate and sodium selenite regulates ROS scavenging in field-grown cowpea plants. Seven Se application rates (0; 2.5; 5; 10; 20; 40 and 60 g ha-1) of each of the two Se forms were applied to plants via the soil. Photosynthetic pigments concentration, gas exchange parameters, lipid peroxidation by malondialdehyde (MDA) concentration, hydrogen peroxide concentration, activity of catalase (CAT, EC:1.11.1.6), glutathione reductase (GR, EC:1.6.4.2), ascorbate peroxidase (APX, EC:1.11.1.11) and Se concentration in leaves and grains were evaluated. In general, Se application led to a decrease in chlorophyll a concentration whilst leading to an increase in chlorophyll b, indicating conservation of total chlorophyll concentration. Application of 2.5 g ha-1 of Se as selenate provided a notable increase in total chlorophyll and total carotenoids compared to the other application rates. Selenate and selenite application decreased lipid peroxidation. However, each Se source acted in a different pathway to combat ROS. While selenate showed more potential to increase activity of APX and GR, selenite showed a higher potential to increase CAT activity. The negative correlation between CAT and GR is indicative that both pathways might be activated under distinct circumstances. The more prominent activity of CAT under high rates of selenite resulted in a negative correlation of this enzyme with chlorophyll a and carotenoids. Both selenate and selenite application increased sucrose and total sugars concentration in leaves of cowpea plants. Overall, these results indicate that application of Se in cowpea under field conditions stimulates distinct pathways to scavenge ROS. This could prove beneficial to mitigate oxidative stress during plant development.

Plant growth-promoting bacteria improve leaf antioxidant metabolism of drought-stressed Neotropical trees.

MAIN CONCLUSION: Plant growth-promoting bacteria association improved the enzymatic and non-enzymatic antioxidant pathways in Neotropical trees under drought, which led to lower oxidative damage and enhanced drought tolerance in these trees. Water deficit is associated with oxidative stress in plant cells and may, thus, negatively affect the establishment of tree seedlings in reforestation areas. The association with plant growth-promoting bacteria (PGPB) is known to enhance the antioxidant response of crops, but this strategy has not been tested in seedlings of Neotropical trees. We evaluated the effects of inoculation with two PGPB (Azospirillum brasilense and Bacillus sp.) on the antioxidant metabolism of Cecropia pachystachya and Cariniana estrellensis seedlings submitted to drought. We measured the activity of antioxidant enzymes and the content of non-enzymatic antioxidants in leaves, and biometrical parameters of the seedlings. In both tree species, drought decreased the activity of antioxidant enzymes and the content of non-enzymatic antioxidant compounds. For C. pachystachya, the enzymatic and non-enzymatic pathways were mostly influenced by A. brasilense inoculation, which enhanced ascorbate peroxidase (APX) and superoxide dismutase activities and positively affected the level of non-enzymatic antioxidant compounds. In C. estrellensis, A. brasilense inoculation enhanced APX activity. However, A. brasilense and Bacillus sp. inoculation had more influence on the non-enzymatic pathway, as both bacteria induced a greater accumulation of secondary compounds (such as chlorogenic acid, gallic acid, rutin and synapic acid) compared to that in non-inoculated plants under drought. For both species, PGPB improved biometrical parameters related to drought tolerance, as specific leaf area and leaf-area ratio. Our results demonstrate that PGPB induced antioxidant mechanisms in drought-stressed Neotropical trees, increasing drought tolerance. Thus, PGPB inoculation provides a biotechnological alternative to improve the success of reforestation programmes.

New insights into cadmium stressful-conditions: Role of ethylene on selenium-mediated antioxidant enzymes.

Cadmium (Cd) contamination has generated an environmental problem worldwide, leading to harmful effects on human health and damages to plant metabolism. Selenium (Se) is non essential for plants, however it can improve plant growth and reduce the adverse effects of abiotic stress. In addition, ethylene may interplay the positive effects of Se in plants. In order to investigate the role of ethylene in Se-modulation of antioxidant defence system in response to Cd-stress, we tested the hormonal mutant Epinastic (epi) with a subset of constitutive activation of the ethylene response and Micro-Tom (MT) plants. For this purpose, Se mineral uptake, Cd and Se concentrations, pigments, malondialdeyde (MDA) and hydrogen peroxide (H2O2) contents, ethylene production, glutathione (GSH) compound, and superoxide dismutase (SOD), ascorbate peroxidase (APX), catalase (CAT), glutathione reductase (GR) and glutathione peroxidase (GSH-Px) activities were analysed in MT and epi plants submitted to 0.5 mM CdCl2 and 1 µM of selenate or selenite. MT plants treated with both Se forms increased growth in the presence or not of 0.5 mM CdCl2, but not change epi growth. Both Se forms reduced Cd uptake in MT plants and cause reverse effect in epi plants. P, Mg, S, K and Zn uptake increased in epi plants with Se application, irrespective to Cd exposure. Chlorophylls and carotenoids contents decreased in both genotypes under Cd exposure, in contrast to what was observed in epi leaves in the presence of Se. When antioxidant enzymes activities were concerned, Se application increased Mn-SOD, Fe-SOD and APX activities. In the presence of Cd, MT and epi plants exhibited decreased SOD activity and increased CAT, APX and GR activities. MT and epi plants with Se supply exhibited increased APX and GR activities in the presence of Cd. Overall, these results suggest that ethylene may be involved in Se induced-defence responses, that triggers a positive response of the antioxidant system and improve growth under Cd stress. These results showed integrative roles of ethylene and Se in regulating the cell responses to stressful-conditions and, the cross-tolerance to stress could be used to manipulate ethylene regulated gene expression to induce heavy metal tolerance.

Se Nanoparticles Induce Changes in the Growth, Antioxidant Responses, and Fruit Quality of Tomato Developed under NaCl Stress.

Nanotechnology represents an opportunity to improve the use of elements in agriculture. Selenium is an element that is beneficial to plants and essential to the human diet. The size of nanoparticles gives them characteristics that can enhance the benefits that selenium provides to plants. The objective of the present study was to determine the effects of selenium nanoparticles on the growth, antioxidant responses, and fruit quality of tomato developed under NaCl stress. Four doses of selenium nanoparticles (1, 5, 10, and 20 mg L-1) under NaCl stress, only NaCl, and a control were evaluated. The results showed that the impact of salinity on the growth of the tomato crop can be reduced with the application of selenium nanoparticles. However, the amount of both enzymatic and non-enzymatic compounds significantly increased in the leaves and fruits of tomato. The results suggest that the application of selenium nanoparticles generated a positive effect against salinity in the tomato crop; moreover, it had a positive impact on the content of beneficial biocompounds for human health in tomato fruits.

The foliar application of a mixture of semisynthetic chitosan derivatives induces tolerance to water deficit in maize, improving the antioxidant system and increasing photosynthesis and grain yield.

Research has shown that chitosan induces plant stress tolerance and protection, but few studies have explored chemical modifications of chitosan and their effects on plants under water stress. Chitosan and its derivatives were applied (isolated or in mixture) to maize hybrids sensitive to water deficit under greenhouse conditions through foliar spraying at the pre-flowering stage. After the application, water deficit was induced for 15 days. Analyses of leaves and biochemical gas exchange in the ear leaf were performed on the first and fifteenth days of the stress period. Production attributes were also analysed at the end of the experiment. In general, the application of the two chitosan derivatives or their mixture potentiated the activities of the antioxidant enzymes superoxide dismutase, catalase, ascorbate peroxidase, glutathione reductase and guaiacol peroxidase at the beginning of the stress period, in addition to reducing lipid peroxidation (malonaldehyde content) and increasing gas exchange and proline contents at the end of the stress period. The derivatives also increased the content of phenolic compounds and the activity of enzymes involved in their production (phenylalanine ammonia lyase and tyrosine ammonia lyase). Dehydroascorbate reductase and compounds such as total soluble sugars, total amino acids, starch, grain yield and harvest index increased for both the derivatives and chitosan. However, the mixture of derivatives was the treatment that led to the higher increase in grain yield and harvest index compared to the other treatments. The application of semisynthetic molecules derived from chitosan yielded greater leaf gas exchange and a higher incidence of the biochemical conditions that relieve plant stress.

Does oxidative stress determine the thermal limits of the regeneration niche of Vriesea friburgensis and Alcantarea imperialis (Bromeliaceae) seedlings?

The predicted environmental changes may be detrimental to initial seedling growth, particularly the expected increase in air temperature. We therefore investigated the thermal limits for growth and development of Vriesea friburgensis and Alcantarea imperialis seedlings in the context of oxidative stress. The optimal temperatures for the growth of V. friburgensis and A. imperialis were 25 and 25-30 °C, respectively. Extreme temperatures (15, 30, or 35 °C) induced oxidative stress in both species with significant accumulation of hydrogen peroxide (H2O2) and nitric oxide (NO). Under oxidative stress, the amount of chlorophyll decreased in both species, more prominently in V. friburgensis, while carotenoid levels dramatically increased in A. imperialis. Notably, the activities of superoxide dismutase, catalase (CAT), and ascorbate peroxidase increased in A. imperialis at extreme temperatures. Similar results were observed for V. friburgensis; however, the activity of CAT remained unaffected regardless of temperature. Seedlings of A. imperialis survived at a wider range of temperatures than V. friburgensis, which had greater than 40% mortality when growing at 30 °C. Overall, precise control of cellular H2O2 and NO levels takes place during the establishment of A. imperialis seedlings, allowing the species to cope with relatively high temperatures. The thermal limits of the fundamental niches of the species investigated, determined based on the ability of seedlings to cope with oxidative stress, were distinct from the realized niches of these species. The results suggest that recruitment success is dependent on the ability of seedlings to handle extreme temperature-triggered oxidative stress, which limits the regeneration niche.

The expression of the genes involved in redox metabolism and hydrogen peroxide balance is associated with the resistance of cowpea [Vigna unguiculata (L.) Walp.] to the hemibiotrophic fungus Colletotrichum gloeosporioides.

Correlations between the transcriptional responses of genes that encode superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX), and peroxiredoxin (Prx) enzymes and Colletotrichum gloeosporioides development in cowpea leaves were assessed. Each of these genes is involved in the redox metabolism and hydrogen peroxide balance. Although electron microscopy revealed that conidia adhered to and germinated on the leaf cuticle, the inoculated cowpea leaves did not show any characteristic anthracnose symptoms. The adhered and germinated conidia showed irregular surfaces and did not develop further. This was apparently due to increased leaf H2O2 levels in response to inoculation with C. gloeosporioides. During the early stages post inoculation, cowpea leaves elevated the H2O2 content and modulated the defense gene expression, as well as associated pathways. During the later stages, the increased expression of the CuZnSODI and CuZnSODII genes suggested an active superoxide dismutation to further elevate H2O2 levels, which indicated that higher H2O2 content may function as a toxic agent that kills the fungus. The second increase in H2O2 production above the threshold level was correlated with the expression of the APXI, CATI, CATII, PrxIIBCD, and PrxIIE genes, which resulted in a coordinated pattern to establish an appropriate balance between H2O2 generation and scavenging. Therefore, appropriate H2O2 content in cowpea leaves inhibited C. gloeosporioides development and maintained intracellular redox homeostasis to avoid uncontrolled programmed cell death and necrosis in cowpea leaves.

Impairment of peroxisomal APX and CAT activities increases protection of photosynthesis under oxidative stress.

Retrograde signalling pathways that are triggered by changes in cellular redox homeostasis remain poorly understood. Transformed rice plants that are deficient in peroxisomal ascorbate peroxidase APX4 (OsAPX4-RNAi) are known to exhibit more effective protection of photosynthesis against oxidative stress than controls when catalase (CAT) is inhibited, but the mechanisms involved have not been characterized. An in-depth physiological and proteomics analysis was therefore performed on OsAPX4-RNAi CAT-inhibited rice plants. Loss of APX4 function led to an increased abundance of several proteins that are involved in essential metabolic pathways, possibly as a result of increased tissue H2O2 levels. Higher photosynthetic activities observed in the OsAPX4-RNAi plants under CAT inhibition were accompanied by higher levels of Rubisco, higher maximum rates of Rubisco carboxylation, and increased photochemical efficiencies, together with large increases in photosynthesis-related proteins. Large increases were also observed in the levels of proteins involved in the ascorbate/glutathione cycle and in other antioxidant-related pathways, and these changes may be important in the protection of photosynthesis in the OsAPX4-RNAi plants. Large increases in the abundance of proteins localized in the nuclei and mitochondria were also observed, together with increased levels of proteins involved in important cellular pathways, particularly protein translation. Taken together, the results show that OsAPX4-RNAi plants exhibit significant metabolic reprogramming, which incorporates a more effective antioxidant response to protect photosynthesis under conditions of impaired CAT activity.

Darkening, damage and oxidative protection are stimulated in tissues closer to the yam cut, attenuated or not by the environment.

BACKGROUND: This study investigated how the activities of the enzymes and metabolites of oxidative metabolism are affected in different regions of cut tissue, associating these changes with the evolution of browning in fresh-cut yam. Samples were collected from yam at 0-5 and 5-10 mm from the cut site. Fresh-cut yams were stored at 5 or 26 °C for days or hours respectively, simulating commercialization, with or without packaging. RESULTS: The results demonstrated that the injury to the yam was a physical inducer of changes in the levels of lipid peroxidation and phenolic compounds and in the activities of superoxide dismutase, catalase, ascorbate peroxidase, polyphenol oxidase and peroxidase. These responses were significant in the tissue closest to the wound but also observed, although less intense, in the more distant tissue. The combined effects of wounding and dehydration during storage intensified the above responses. Conversely, refrigeration attenuated the transmission of the wounding response through the adjacent tissue. CONCLUSION: The results of this work provide the first evidence in cut yam roots that the membrane degradation products, enzymes involved in oxidative protection and enzymes that modulate the oxidation of phenolic compounds are intertwined mechanisms that cause tissue darkening. © 2018 Society of Chemical Industry.

Proteomic and physiological approaches reveal new insights for uncover the role of rice thylakoidal APX in response to drought stress.

Chloroplast APX isoforms display controversial roles as H2O2 scavengers and signaling players in response to abiotic stress and conclusive results are lacking. We tested the hypothesis that thylakoidal APX displays an important role for drought tolerance, especially by regulating abundance of essential protein species. For this, OsApx8 RNAi-silenced rice (apx8) and non-transformed plants (NT) were exposed to mild water deficit. The drought-sensitivity in apx8 plants was revealed by decreases in shoot growth, relative water content and photosynthesis, which was accompanied by increased membrane damage, all compared to NT plants. This higher sensitivity of apx8 plants to mild drought stress was also related to a lower accumulation of important protein species involved in several metabolic processes, especially photosynthesis, photorespiration and redox metabolism. Despite apx8 plants have displayed an effective induction of compensatory antioxidant mechanisms in well-watered conditions, it was not enough to maintain H2O2 homeostasis and avoid oxidative and physiological disturbances under mild drought conditions. Thus, thylakoidal APX is involved in several phenotypic modifications at proteomic profile level, possibly via a H2O2-induced signaling mechanism. Consequently, this APX isoform is crucial for rice plants effectively cope with a mild drought condition. BIOLOGICAL SIGNIFICANCE: This work provides for the first time an integrative study involving proteomic, physiological and biochemical analyses directed to elucidation of thylakoidal APX roles for drought tolerance in rice plants. Our data reveal that this enzyme is crucial for maintaining of growth and photosynthesis under mild water deficit conditions. This essential role is related to maintaining of H2O2 homeostasis and accumulation of essential proteins involved in several important metabolic pathways. Remarkably, for drought resistance was essential the accumulation of proteins involved with metabolism of photosynthesis, signaling, carbohydrates, protein synthesis/degradation and stress. These results can contribute to understand the role of chloroplast ascorbate peroxidases in drought tolerance, highlighting the physiological importance of key proteins in this process.