Cleberson Ribeiro

Cadmium-induced oxidative stress and antioxidative enzyme response in water hyacinth and salvinia

The reactive oxygen species generation, lipid peroxidation and antioxidative enzyme response of water hyacinth and salvinia to Cd were evaluated. Cadmium was absorbed/accumulated mainly in the roots, but significant amounts also translocated to the leaves. No Cd effect on dry weight was detected, although toxicity symptoms were visible. Superoxide and H2O2 concentrations increased, in addition to lipid peroxidation in both species, especially in the leaves of salvinia. In general, antioxidative enzyme activities were reduced in both species following Cd treatment, especially in salvinia. Glutathione peroxidase (GPX, EC 1.11.1.9) activity decreased in water hyacinth but increased in salvinia. Glutathione S-transferase (GST, EC 2.5.1.18) activity increased in the leaves but decreased in the roots of both species. So, Cd induced ROS generation/accumulation, but the antioxidative enzymes were not able to combat the Cd-induced oxidative injury in these two species. Nevertheless, water hyacinth consistently showed a higher tolerance to Cd than salvinia.

Impact of iron toxicity on oxidative metabolism in young Eugenia uniflora L. plants

In excess, iron can induce the production and accumulation of reactive oxygen species (ROS), causing oxidative stress. The objective of this work was to evaluate the impact of toxic concentrations of iron (Fe) on the antioxidative metabolism of young Eugenia uniflora plants. Forty-five-day-old plants grown in Hoagland nutrient solution, pH 5.0, were treated with three Fe concentrations, in the form of FeEDTA, during three periods of time. At the end of the treatment, the plants were harvested and relative growth rate, iron content, lipid peroxidation and enzymes and metabolites of the antioxidative metabolism were determined. Iron-treated plants showed higher iron contents, reduced relative growth rates and iron toxicity symptoms in both leaves and roots. There was an increase in lipid peroxidation with increasing Fe, only in the leaves. The enzymatic activities of superoxide dismutase (SOD) and glutathione reductase (GR) increased with increasing Fe concentration and treatment exposure time. The activities of catalase (CAT), peroxidase (POX) and ascorbate peroxidase (APX) also increased with increasing Fe concentration but decreased with increasing treatment exposure time. Glutathione peroxidase activity (GPX) decreased with increasing Fe concentration and exposure time. The ascorbate (AA) and reduced glutathione (GSH) contents and the AA/DHA and GSH/GSSG ratios, in general, increased with increasing Fe concentration and treatment exposure time. The results indicate that under toxic levels of Fe, young E. uniflora plants suffer increased oxidative stress, which is ameliorated through changes in the activities of antioxidative enzymes and in the contents of the antioxidants AA and GSH.

Corrigendum: The Involvement of Nitric Oxide in Integration of Plant Physiological and Ultrastructural Adjustments in Response to Arsenic

High arsenic (As) concentrations are toxic to all the living organisms and the cellular response to this metalloid requires the involvement of cell signaling agents, such as nitric oxide (NO). The As toxicity and NO signaling were analyzed in Pistia stratiotes leaves. Plants were exposed to four treatments, for 24 h: control; SNP [sodium nitroprusside (NO donor); 0.1 mg L-1]; As (1.5 mg L-1) and As + SNP (1.5 and 0.1 mg L-1, respectively). The absorption of As increased the concentration of reactive oxygen species and triggered changes in the primary metabolism of the plants. While photosynthesis and photorespiration showed sharp decrease, the respiration process increased, probably due to chemical similarity between arsenate and phosphate, which compromised the energy status of the cell. These harmful effects were reflected in the cellular structure of P. stratiotes, leading to the disruption of the cells and a possible programmed cell death. The damages were attenuated by NO, which was able to integrate central plant physiological processes, with increases in non-photochemical quenching and respiration rates, while the photorespiration level decreased. The increase in respiratory rates was essential to achieve cellular homeostasis by the generation of carbon skeletons and metabolic energy to support processes involved in responses to stress, as well to maintaining the structure of organelles and prevent cell death. Overall, our results provide an integrated view of plant metabolism in response to As, focusing on the central role of NO as a signaling agent able to change the whole plant physiology.

Antioxidant system response induced by aluminum in two rice cultivars

The antioxidant defense system response was evaluated in two rice cultivars (Oryza sativa L.), Fernandes (CNA-1158) and Maravilha (CNA-6843-1), treated with toxic levels of aluminum. After exposure to aluminum we determined plant growth, H2O2 and O2•- contents, lipid peroxidation, antioxidant enzymes activities and ascorbate and dehydroascorbate contents. Al predominantly accumulated in roots of both cultivars but it reduced root and shoot growth only in the Maravilha cultivar. Treatment with aluminum resulted in a reduction of 84 and 60% in the levels of H2O2 in the roots of the cultivars Maravilha and Fernandes, respectively, and of 26% in the levels of O2•- only in the roots of Fernandes cultivar. Increased lipid peroxidation was observed only in the roots of the Maravilha cultivar. In general, the antioxidant enzyme activities were higher in roots and increased in the presence of aluminum, especially in the Fernandes cultivar. The levels of ascorbate were higher in leaves and increased with aluminum treatment, while dehydroascorbate decreased in roots of both cultivars after aluminum treatment. However, the ascorbate/dehydroascorbate ratio increased in the roots of both cultivars after treatment with aluminum. Ascorbate, dehydroascorbate and ascorbate/dehydroascorbate levels found here point to an efficient regeneration of ascorbate, essential for the homeostasis of cellular metabolites involved in reactive oxygen species removal by rice plants treated with aluminum. Therefore, the higher tolerance of Fernandes to aluminum relative to Maravilha cultivar may be the result of better growth of the root system and shoots, higher antioxidant enzyme activities and a best use/regeneration of ascorbate.

Arsenic toxicity: cell signalling and the attenuating effect of nitric oxide in Eichhornia crassipes

Nitric oxide (NO) is an important molecule involved in the perception of stress induced by toxic compounds such as arsenic (As). The present study investigated the role of NO applied as sodium nitroprusside (SNP) in cell signalling and the ability of NO to attenuate the toxic effects of As (in the form of sodium arsenate) in water hyacinth (Eichhornia crassipes). Water hyacinth plants were collected and assigned to one of the following treatments: control; 100 μM SNP; 20 μM As; or 20 μM As + 100 μM SNP. The plants remained under these conditions for 0, 4, 12, and 24 h. After each time interval, the plants were collected and As absorption, production of reactive oxygen species (ROS), integrity of membranes, and antioxidant enzyme activities were evaluated. The plants were able to absorb and accumulate large amounts of As, even after only four hours of exposure to the pollutant. The absorption and bioaccumulation factor of As was even greater when plants were exposed to both As and SNP. The accumulation of As triggered increases in ROS production and cell membrane damage. In the presence of SNP, the tolerance index to As increased and damage was mitigated. Therefore, from the present work, it was possible to conclude that exogenous NO influenced the ability of plants to tolerate As; this finding has implications for phytoremediation in areas contaminated by As.

Proline levels, oxidative metabolism and photosynthetic pigments during in vitro growth and acclimatization of Pitcairnia encholirioides L.B. Sm. (Bromeliaceae).

This study aimed to evaluate the variation in the levels of proline, oxidative metabolism and photosynthetic pigments in plants of Pitcairnia encholirioides grown in vitro under different conditions and after acclimatization. The analyses were performed after 150 days of in vitro cultivation in MS media supplemented with 10 µM GA3 or 0.2 µM NAA, sucrose at 15 or 30 g L-1, in test tubes which allowed gas exchange or in a hermetically sealed system, and 180 days after acclimatization. The in vitro maintenance in hermetically sealed flasks, with GA3 and 15 g L-1 sucrose had adverse metabolic effects, which was demonstrated by the lower proline and photosynthetic pigments accumulation and by the increase in antioxidant enzymes activities. After acclimatization, differences for proline and photosynthetic pigments were no longer found and the enzymatic activities ranged unevenly. The results suggest that the in vitro cultivation in media with 0.2 µM NAA and 30 g L-1 sucrose, in test tubes capped with closures which allowed gas exchange, is more suitable for micropropagation of P. encholirioides, providing a prolonged maintenance of in vitro cultures and plantlets with superior quality for ex vitro development.

Efeito do alumínio sobre a absorção, o acúmulo e o fracionamento do fósforo em sorgo

ABSTRACTALUMINUM EFFECT ON UPTAKE, ACCUMULATION AND FRACTIONATIONOF PHOSPHORUS IN SORGHUM The objective of this work was to evaluate Al effect on uptake, accumulation and fractionation ofP in two sorghum cultivars. Plants were treated with toxic levels of Al during ten days and then theywere harvested and growth, dry matter yield, Al and total P contents and concentrations of the variousP forms in the two parts of the plants were determined. Aluminum effect on P uptake was also evaluatedin intact plants. Aluminum reduced the growth of the seminal root and dry matter yield in roots andtops of both cultivars, especially in the sensitive one. Aluminum and P contents increased in roots butdid not change in the top of both cultivars. Phosphate uptake by roots, however, decreased in the presenceof Al in both cultivars, especially in the sensitive one. Aluminum, in general, changed concentrations ofall soluble (P i e P org ) and insoluble P forms (P RNA e P residual ), except of the P LIP form. Some of thesemodifications seem to be important and may be related to Al tolerance mechanism in sorghum.Key words: aluminum tolerance, Sorghum bicolor, inorganic phosphate, organic phosphate.(

Effects of aluminum on the elongation and external morphology of root tips in two maize genotypes

The objective of this study was to evaluate the effects of toxic levels of aluminum (Al) on the growth and external morphology of root tips in two maize genotypes with differential Al tolerance. The maize genotypes UFVM-100 (Al-sensitive) and UFVM-200 (Al-tolerant) were treated with 0 and 50 mM Al in a 0.5 mM CaCl2 solution at pH 4.5; root elongation, Al content and the external morphology of the root tips were evaluated. Chemical analysis, hematoxylin staining and energy dispersive X-ray spectrometry (EDS) showed greater Al accumulation in the root tips of the UFVM-200 genotype. The inhibition of root elongation in the UFVM-100 genotype, however, was much stronger than in the UFVM-200 genotype. Both maize genotypes exhibited visible and intense alterations in external micromorphology of the root tip, especially the Al-sensitive UFVM-100 genotype. Scanning electron micrographs showed intense cell disorganization and transverse ruptures of the protodermic and outer cortex layers of the cells in both genotypes. The ruptures were deeper and wider and reached the inner cortex layers in the UFVM-100 genotype. The EDS analysis showed that, in addition to Al accumulation, there was a proportional increase in the P concentration in the root tips of the UFVM-200 genotype. This is indicative of possible precipitation and/or immobilization of Al in the root tip apoplast of this genotype, which contributes to symplastic detoxification.

Citric acid secretion induced by aluminum in two Stylosanthes species

Secretion of organic acids (OAs) by roots has been suggested to be an important mechanism of Al resistance in many species. In Stylosanthes, the participation of OAs in the mechanism of Al resistance is poorly understood. We aimed to study the production and secretion of OAs by two Brazilian Stylosanthes species with different Al resistance. Stylosanthes capitata and S. guianensis were treated with Al at different concentrations in 0.5 mM CaCl2 (pH 4.0), and then root elongation, Al and OA content, OA secretion into the external solution, and the activity of citrate synthase (CS) were measured. Al-induced secretion of citric acid was also evaluated in the presence of protein synthesis and anion channel inhibitors. S. guianensis accumulated lower amounts of Al in its roots and displayed less inhibition of root elongation compared to S. capitata. Citric and malic acids were the most abundant OAs in the roots, and their content decreased with the Al treatment, except for citric acid in S. guianensis. Citric acid was the only OA secreted into the nutrient solution by the Al-treated plants of both species, but more by S. guianensis. Citrase synthase activity decreased in S. capitata but increased in S. guianensis with the Al treatment, and it may have a crucial role in the maintenance of citric acid content in the roots of S. guianensis. The use of anion channel and protein synthesis inhibitors reveal that anion channels were likely involved in the secretion of citric acid, and channel protein transcription was up-regulated by exposure to Al in Stylosanthes.

Oxidative stress triggered by arsenic in a tropical macrophyte is alleviated by endogenous and exogenous nitric oxide

Nitric oxide (NO) plays important role in alleviating abiotic stresses in plants, including those caused by arsenic (As). Here, we examined the effects of endogenous and exogenous NO in Spirodela intermedia W. Koch (Lemnaceae) under As exposure. For this purpose, we evaluated the As content, reactive oxygen species (ROS) levels, membrane damage and enzymatic antioxidant system. The levels of endogenous NO and the activity of nitrate reductase (NR) were also addressed. The As treatment triggered the production of high endogenous levels of NO and a pronounced activation of the antioxidant enzymes; however, it was not sufficient to completely avoid the increment in ROS content and membrane damage. In contrast, exogenous NO decreased the As levels in plants exposed to As and NO donor, mitigating the ROS production and membrane damage, while maintaining a lower activity of the antioxidant enzymes compared with As-treated plants. Exogenous NO further downregulated the NR activity by a negative feedback, while As boosted the NR activity, consistent with the high endogenous levels of NO observed upon As treatment. Our results suggest that both endogenous and exogenous NO play critical roles in alleviating the As-induced oxidative stress in S. intermedia by reducing As uptake, and possibly by acting as an antioxidant molecule.