Copper oxychloride fungicide and its effect on growth and oxidative stress of potato plants.

Excess copper in plants causes physiological alterations that lead to crop productivity losses. However, cupric fungicides have been utilized in the control of Alternaria solani and Phytophthora infestans fungi, which cause early blight and late blight in potato, respectively. Thus, this study aimed to investigate the effect of different copper oxychloride levels on potato plants through some biochemical and physiological parameters. The fungicide was applied at the recommended level (2.50gL(-1)), at a reduced level (1.25gL(-1)), and at 5.00gL(-1), to simulate spraying in the field twice during the same period with the recommended level. The results revealed that superoxide dismutase (SOD, EC 1.15.1.1) protected plants against oxidative stress at the beginning of the cycle since lipoperoxide levels were low in that period. In addition, increased SOD activity positively correlated with increased usable leaf area for photosynthesis (leaf area ratio, LAR), photosynthetic effectiveness (net assimilation rate, NAR), and growth relative to pre-existing dry matter (relative growth rate, RGR). Concomitantly, there was a negative correlation between lipoperoxide levels and LAR and RGR. Plants randomly sprayed twice in the same period with the level recommended for potato crop protection in the field do not present damage regarding their development. However, additional studies are needed in order to reduce the use of copper fungicides in the control of early and late blight in potato crop production, then decreasing the release of copper in the environment.

Differential responses between mature and young leaves of sunflower plants to oxidative stress caused by water deficit / Diferentes respostas entre folhas adultas e jovens de plantas de girassol ao estresse oxidativo causado pelo déficit hídrico

The effects of water stress and rehydration on leaf gas exchange characteristics along with changes in lipid peroxidation and pirogalol peroxidase (PG-POD) were studied in mature and in young leaves of sunflower (Helianthus annuus L.), which were grown in a greenhouse. Water stress reduced photosynthesis (Pn), stomatal conductance (g s), and transpiration (E) in both young and mature leaves. However, the amplitude of the reduction was dependent on leaf age. The intercellular CO2 concentration (Ci) was increased in mature leaves but it was not altered in young leaves. Instantaneous water use efficiency (WUE) in mature stressed leaves was reduced when compared to control leaves while in young stressed leaves it was maintained to the same level as the control. After 24h of rehydration, most of the parameters related to gas exchange recovered to the same level as the unstressed plants except gs and E in mature leaves. Water stress did not activated PG-POD independently of leaf age. However, after rehydration the enzyme activity was increased in mature leaves and remained to the same as the control in young leaves. Malondialdehyde (MDA) content was increased by water stress in both mature and young leaves. The results suggest that young leaves are more susceptible to water stress in terms of gas exchange characteristics than mature leaves although both went through oxidative estresse.

Os efeitos do estresse hídrico e da reidratação nas trocas gasosas juntamente com alterações na lipoperoxidação e atividade da pirogalol peroxidase (PG-POD) foram estudados em folhas adultas e jovens de plantas de girassol (Helianthus annuus L.) cultivadas em casa de vegetação. O estresse hídrico reduziu a fotossíntese (P N), a condutância estomática (g s) e a transpiração (E) nas folhas adultas e jovens. No entanto, a amplitude da redução foi dependente da idade da folha. A concentração intercelular de CO2 (Ci) aumentou nas folhas adultas, mas não apresentou alteração nas folhas jovens sob condições de estresse hídrico. A eficiência instantânea do uso da água (WUE) nas folhas adultas e estressadas foi reduzida em comparação com o controle, enquanto que, nas folhas jovens e estressadas, não foi alterada. Após 24h de reidratação, as folhas apresentaram valores semelhantes aos dos controles em todos os parâmetros relacionados com as trocas gasosas, exceto gs e E nas folhas adultas. O estresse hídrico não ativou a PG-POD, independente da idade da folha, e após a reidratação a atividade da PG-POD foi aumentada apenas nas folhas adultas. O estresse hídrico aumentou o teor de malondialdeído (MDA) nas folhas adultas e jovens. Os resultados sugerem que as folhas jovens são mais susceptívies ao déficit hídrico em termos de trocas gasosas que as folhas adultas, embora ambas tenham sido expostas ao estresse oxidativo.

Alcoholism and alcohol abstinence: N-acetylcysteine to improve energy expenditure, myocardial oxidative stress, and energy metabolism in alcoholic heart disease.

Alcoholism has been associated with a wide range of pathologic conditions, including alcoholic heart disease (AHD). Because AHD may be associated with oxidative stress, antioxidant compounds, such as N-acetylcysteine (NAC) could be useful to control the damage done by alcohol (ethanol) consumption. To investigate the NAC effects on alcoholism and alcohol abstinence, initially, 30 male Wistar rats were divided into two groups: (C, N=6) given standard chow and water; (E, N=24) receiving standard chow and aqueous ethanol solution in semi-voluntary research. After 30 days of ethanol-exposure, (E) group was divided into four subgroups (N=6/group):(E-E) continued drinking 30% ethanol-solution; (E-NAC) drinking ethanol-solution containing 2g/L NAC; (AB) changed ethanol solution to water; (AB-NAC) changed ethanol to aqueous solution of 2g/L NAC. After 15 days of the E-group division, E-E rats had lower body weight and feed efficiency, as well as higher energy-expenditure resting metabolic rate (RMR)/body weight and VO(2) consumption/surface area. These calorimetric changes were reflected on the cardiac tissue. E-E rats had higher heart weight/body weight ratio and myocardial lipid hydroperoxide (LH), indicating AHD with hypertrophy and oxidative stress. Myocardial superoxide dismutase was higher, whereas glutathione-peroxidase (GSH-peroxidase) was lower in E-E rats than in C. The higher myocardial hydroxyacyl coenzyme-A dehydrogenase (OHADH), OHADH/citrate synthase (CS), and lactate dehydrogenase (LDH)/CS in E-E rats indicated higher fatty acid degradation relative to aerobic metabolism predisposing the lipotoxicity. AB rats had lower RMR/body weight than E-E, normalized myocardial oxidative stress, and energy metabolism. E-NAC and AB-NAC had lower RMR/body weight, myocardial LH, LDH/CS, and higher GSH-peroxidase than E-E and AB, respectively, demonstrating lower oxidative stress and higher myocardial carbohydrate oxidation. In conclusion, the present study brought new insights on alcohol consumption and AHD because ethanol-exposure enhanced energy-expenditure and induced a number of calorimetric changes, which were reflected in body weight and myocardial lipotoxicity. NAC preventing ethanol-induced calorimetric changes and reducing myocardial oxidative stress enhanced carbohydrate oxidation, thus optimizing myocardial energy metabolism in both alcoholic and abstinence condition.

Ultraviolet-B and water stress effects on growth, gas exchange and oxidative stress in sunflower plants.

The effects and interaction of drought and UV-B radiation were studied in sunflower plants (Helianthus annuus L. var. Catissol-01), growing in a greenhouse under natural photoperiod conditions. The plants received approximately 1.7 W m(-2) (controls) or 8.6 W m(-2) (+UV-B) of UV-B radiation for 7 h per day. The UV-B and water stress treatments started 18 days after sowing. After a period of 12 days of stress, half of the water-stressed plants (including both UV-B irradiated or non-irradiated) were rehydrated. Both drought and UV-B radiation treatments resulted in lower shoot dry matter per plant, but there was no significant interaction between the two treatments. Water stress and UV-B radiation reduced photosynthesis, stomatal conductance and transpiration. However, the amplitude of the effects of both stressors was dependent on the interactions. This resulted in alleviation of the negative effect of drought on photosynthesis and transpiration by UV-B radiation as the water stress intensified. Intercelluar CO(2) concentration was initially reduced in all treatments compared to control plants but it increased with time. Photosynthetic pigments were not affected by UV-B radiation. Water stress reduced photosynthetic pigments only under high UV-B radiation. The decrease was more accentuated for chlorophyll a than for chlorophyll b. As a measure for the maximum efficiency of photosystem II in darkness F (v)/F (m) was used, which was not affected by drought stress but initially reduced by UV-B radiation. Independent of water supply, UV-B radiation increased the activity of pirogalol peroxidase and did not increase the level of malondialdehyde. On the other hand, water stress did not alter the activity of pirogalol peroxidase and caused membrane damage as assessed by lipid peroxidation. The application of UV-B radiation together with drought seemed to have a protective effect by lowering the intensity of lipid peroxidation caused by water stress. The content of proline was not affected by UV-B radiation but was increased by water stress under both low and high UV-B radiation. After 24 h of rehydration, most of the parameters analyzed recovered to the same level as the unstressed plants.

Eucalyptus ESTs associated with resistance to herbicide inhibitors of aromatic and branched-chain amino acid synthesis

Herbicides inhibit enzymatic systems of plants. Acetolactate synthase (ALS, EC = 4.1.3.18) and 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS, EC 2.5.1.19) are key enzymes for herbicide action. Hundreds of compounds inhibit ALS. This enzyme is highly variable, enabling the selective control of weeds in a number of crops. Glyphosate, the only commercial herbicide inhibiting EPSPS is widely used for non-selective control of weeds in many crops. Recently, transgenic crops resistant to glyphosate were developed and have been used by farmers. The aim of this study was the data mining of eucalypt expressed sequence tags (ESTs) in the FORESTs Genome Project database (https://forests.esalq.usp.br) related to these enzymes. Representative amino acid sequences from the NCBI database associated with ALS and EPSPS were blasted with ESTs from the FORESTs database using the tBLASTx option of the blast tool. The best blasting reads and clusters from FORESTs, represented as nucleotide sequences, were blasted back with the NCBI database to evaluate the level of similarity with available sequences from different species. One and seven clusters were identified as showing high similarity with EPSPS and ALS sequences from the literature, respectively. The alignment of EPSPS sequences allowed the identification of conserved regions that can be used to design specific primers for additional sequencings

Eucalyptus ESTs corresponding to the enzyme glutamine synthetase and the protein D1, sites of action of herbicides that cause oxidative stress

This work was aimed at locating Eucalyptus ESTs corresponding to the GS enzyme (Glutamine Synthetase, EC = 6.3.1.2) and to the D1 protein, which are directly related to resistance to herbicides that promote oxidative stress. Glutamine Synthetase corresponds to the site of action of the herbicide glufosinate. Herbicides that belong to groups such as ureas, uracils, triazines and triazinones act on the D1-Qb complex (receptor of electrons from the Photosystem II) by inactivating it. The clusters EGEQRT3302E01.g, EGEQRT3001F12.b; EGEZLV1203B04.g; EGBGFB1211H06.g and EGEZLV1205F09.g enclosed complete sequences (with 356 amino acids) of the Glutamine Synthetase enzyme. The cluster EGEQSL1054G06.g is a consensus of four reads and enclosed a complete sequence of D1 Protein (with 353 amino acids). The comparison of the sequences of Protein D1 from different species showed that the substitutions of serine (S) by glycine (G) or serine (S) by threonine (T) at the position 264 could produce plants resistant to herbicides that act on electron flow on Photosystem II. The sequence of amino acids corresponding to the cluster EGEQSL1054G06.g had a serine in position 264 indicating sensitivity of the Eucalyptus plants to herbicides that act on this site

Eucalyptus ESTs corresponding to the protoporphyrinogen IX oxidase enzyme related to the synthesis of heme, chlorophyll, and to the action of herbicides

This work was aimed at locating Eucalyptus ESTs corresponding to the PROTOX or PPO enzyme (Protoporphyrinogen IX oxidase, E.C. 1.3.3.4) directly related to resistance to herbicides that promote oxidative stress, changing the functionality of this enzyme. PROTOX, which is the site of action of diphenyl-ether (oxyfluorfen, lactofen, fomesafen), oxadiazole (oxadiazon and oxadiargyl), and aryl triazolinone (sulfentrazone and carfentrazone) herbicides, acts on the synthesis route of porphyrins which is associated with the production of chlorophyll a, catalases, and peroxidases. One cluster and one single read were located, with e-values better than e-70, associated to PROTOX. The alignment results between amino acid sequences indicated that this enzyme is adequately represented in the ESTs database of the FORESTs project

Eucalyptus ESTs related to genes for oxidative stress.

Oxidative stress generating active oxygen species has been proved to be one of the underlying agents causing tissue injury after the exposure of Eucalyptus (Eucalyptus spp.) plants to a wide variety of stress conditions. The objective of this study was to perform data mining to identify favorable genes and alleles associated with the enzyme systems superoxide dismutase, catalase, peroxidases, and glutathione S-transferase that are related to tolerance for environmental stresses and damage caused by pests, diseases, herbicides, and by weeds themselves. This was undertaken by using the eucalyptus expressed-sequence database (https//forests.esalq.usp.br). The alignment results between amino acid and nucleotide sequences indicated that the studied enzymes were adequately represented in the ESTs database of the FORESTs project.