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Item Aluminum-induced citric acid secretion is not the sole mechanism of Al-resistance in maize(Acta Physiologiae Plantarum, 2016-12) Cardoso, Flávio Barcellos; Cambraia, José; Oliveira, Juraci Alves de; Ribeiro, Cleberson; Souza, Leandro Torres de; Braun, Heder; DaMatta, Fábio MuriloAluminum-induced citric acid (CA) root secretion is a widely accepted mechanism to explain Al-resistance in maize. Nonetheless, several aspects of this mechanism remain controversial. In this study, we used paclobutrazol (PBZ), a plant growth retardant, to gain new insights into the relationship between Δ5-sterol composition, membrane permeability, (PM) H+-ATPase activity and CA secretion in an Al-sensitive (UFVM-100) and Al-resistant (UFVM-200) maize genotypes challenged with Al. The Al-sensitive genotype displayed greater concentrations of Al in the root tips and greater inhibition of root elongation (RE), which was accompanied by greater electrolyte leakage and greater reduction in the Δ5-sterols content after Al treatment. CA secretion by roots increased in both genotypes after Al treatment but to a greater extent in the Al-resistant genotype. The (PM) H+-ATPase activity was down-regulated in the sensitive cultivar and up-regulated in its resistant counterpart upon Al treatment. A significant correlation between (PM) H+-ATPase activity and CA secretion was observed, but only in the Al-resistant genotype. Upon adding PBZ to the Al-treated plants, differences in the RE and Δ5-sterol composition between the maize genotypes were fully abolished, whereas genotypic differences in CA secretion and (PM) H+-ATPase activity were reduced but not completely eliminated. Taken together, this information suggests the existence of other processes or mechanisms operating in the Al resistance in these two maize genotypes.Item Antioxidant system response induced by aluminum in two rice cultivars(Brazilian Journal of Plant Physiology, 2012-04) Ribeiro, Cleberson; Cambraia, José; Fonseca Júnior, Élcio Meira da; Peixoto, Paulo Henrique PereiraThe 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.Item Bacteriophage isolated from sewage eliminates and prevents the establishment of Escherichia Coli Biofilm(Advanced Pharmaceutical Bulletin, 2018-02-20) Ribeiro, Karla Veloso Gonçalves; Ribeiro, Cleberson; Dias, Roberto Sousa; Cardoso, Silvia Almeida; Paula, Sérgio Oliveira de; Zanuncio, Jose Cola; Oliveira, Leandro Licursi deBiofilm growth exerts a negative impact on industry and health, necessitating the development of strategies to control. The objective of this work was study the lytic activity of the phage isolated from the sewage network in the formation and degradation of Escherichia coli biofilms. E. coli cultures were incubated in 96-well polystyrene microplates under controlled conditions to evaluate the biofilm formation. The E. coli cultures and established biofilms were treated with the suspensions of the vB_EcoM-UFV017 (EcoM017) bacteriophage obtained from sewage for 24 hours. The E. coli bacterial density was measured using absorbance at 600 nm and the biofilms were measured by crystal violet staining. Polystyrene coupons were used as support for Scanning Electron Microscopy and Confocal Microscopy to evaluate biofilm formation. The E. coli strains formed biofilms in polystyrene microplates after 48 hours’ incubation. The highest EcoM017 phage titer, in the prevention and degradation experiments, reduced the bacterial growth and the quantity of biofilm formed by E. coli in 90.0% and 87.5%, respectively. The minimum dose capable of reducing the biofilms of this bacterium was 101 PFU/mL after 24 hours. The preformed E. coli biofilm mass was reduced 79% post exposure to the phage in the degradation assay. Microscopic analysis confirmed the results obtained in the plates assays. The EcoM017 phage prevented biofilm formation and degraded the E. coli-established ones. The EcoM017 phage isolated from sewage can reduce bacterial attachment and lyse the E. coli associated biofilm cells, offering biotechnological potential applicability for this phage.Item Crescimento, morfologia e mudanças no sistema antioxidativo em raízes de arroz, submetidas ao alumínio(Universidade Federal de Viçosa, 2011-04-28) Ribeiro, Cleberson; Silva, Marco Aurélio Pedron e; http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4790898P8; Cano, Marco Antonio Oliva; http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4787546T4; Cambraia, José; http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4783868U6; http://lattes.cnpq.br/5023387764069051; Peixoto, Paulo Henrique Pereira; http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4780011D1; Azevedo, Aristéa Alves; http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4787822Y7Em solos ácidos, a toxicidade do alumínio (Al) é um dos principais fatores que limita o crescimento e o desenvolvimento das plantas. A exposição de plantas a níveis tóxicos desse elemento pode elevar a produção de espécies reativas de oxigênio (ROS) e, consequentemente, induzir estresse oxidativo. Para evitar o acúmulo excessivo de ROS, as plantas utilizam sistemas de defesa antioxidativo envolvendo certos metabólitos e diversas enzimas antioxidativas. O presente trabalho objetivou avaliar alguns danos causados pelo Al sobre o crescimento e a morfologia das raízes e as respostas do sistema ascorbato/glutationa em dois cultivares de arroz: Fernandes (CNA-1158) e Maravilha (CNA-6843-1), considerados tolerante e sensível ao Al, respectivamente. Após nove dias de tratamento das plantas com Al nas concentrações de 0 e 1 mM foram avaliados os efeitos desse metal sobre o crescimento e morfologia das raízes, sobre os teores das ROS, ascorbato (AA), desidroascorbato (DHA), glutationa reduzida (GSH) e glutationa oxidada (GSSG) e sobre as atividades das enzimas dismutase do superóxido (SOD), peroxidase do ascorbato (APX), catalase (CAT), peroxidase (POX), peroxidase da glutationa (GPX), redutase da glutationa (GR), γ-glutamilcisteinil sintase (γ-GCS), glutationa Stransferase (GST), redutase do monodesidroascorbato (MDHAR) e redutase do desidroascorbato (DHAR). O cultivar Maravilha apresentou maior acúmulo de Al e menor crescimento de suas raízes do que o cultivar Fernandes. O Al acumulou-se especialmente nos ápices radiculares dos dois cultivares, mas as raízes do cultivar Maravilha, tornaram-se mais grossas do que as do cultivar Fernandes. O acúmulo de Al provocou danos estruturais, com ruptura das células da coifa, alterando a morfologia externa dos ápices radiculares dos dois cultivares de arroz, principalmente no cultivar Maravilha. O cultivar Fernandes apresentou menor teor de ROS e menor grau de peroxidação de lipídios do que o cultivar Maravilha, caracterizando sua maior tolerância ao Al. O cultivar Fernandes apresentou, também, maior atividade das enzimas SOD, APX, CAT e POX e maior regeneração do AA em suas raízes, indicativo de um mecanismo mais eficiente na eliminação do excesso vii de ROS produzido durante o estresse oxidativo causado por Al. As atividades das enzimas γ-GCS e GR, envolvidas na síntese e regeneração da glutationa, respectivamente, aumentaram nas raízes do cultivar Fernandes, enquanto no cultivar Maravilha, apenas a enzima γ-GCS apresentou aumento em sua atividade no 9º dia de tratamento com Al. A atividade da GPX aumentou nas raízes dos dois cultivares tratados com Al, mas no cultivar Maravilha o efeito do Al foi menos intenso e mais tardio. O cultivar Fernandes, na presença de Al, foi capaz de manter teores de GSH e da relação GSH/GSSG mais elevados e, portanto, compatíveis com a operação de melhor tampão de redox do que no cultivar Maravilha. Portanto, a maior parte da capacidade do arroz tolerar o estresse oxidativo causado por Al parece ser resultado da operação sincronizada de sistemas enzimáticos e não-enzimáticos relacionados com o metabolismo do ascorbato e da glutationa. Tanto a atividade das enzimas quanto os teores de ascorbato e, ou de glutationa indicam maior eficiência do sistema ascorbato/glutationa no cultivar Fernandes do que no cultivar Maravilha.Item Efeito do alumínio sobre a absorção, o acúmulo e o fracionamento do fósforo em sorgo(Bragantia, 2008-10) Pereira, Jamil Morais; Cambraia, José; Fonseca Júnior, Élcio Meira da; Ribeiro, ClebersonO trabalho teve como objetivo estudar o efeito do Al sobre a absorção, o acúmulo e o fracionamento do P em duas cultivares de sorgo. As plantas foram expostas a níveis tóxicos de Al durante dez dias e, então, colhidas e determinados o crescimento em tamanho e produção de massa seca, os teores de Al e de P total e as diversas formas de P nas duas partes das plantas. Avaliou-se, também, o efeito do Al sobre a absorção de P pelas raízes de plantas intactas. O Al reduziu o crescimento da raiz seminal e a produção de matéria seca de raízes e parte aérea nas duas cultivares, especialmente na sensível. Os teores de Al e de P total aumentaram nas raízes, mas não foram modificados na parte aérea nas duas cultivares. A absorção de P, entretanto, decresceu na presença de Al nas duas cultivares, principalmente na sensível. O Al, de modo geral, modificou as concentrações das várias formas de P solúvel (Pi e Porg) e insolúvel (PRNA e Presidual), exceto a da forma PLIP. Algumas dessas modificações parecem ser importantes e podem estar relacionadas com o mecanismo de tolerância ao Al em sorgo.Item Effects of aluminum on the elongation and external morphology of root tips in two maize genotypes(Bragantia, 2016-01) Souza, Leandro Torres de; Cambraia, José; Ribeiro, Cleberson; Oliveira, Juraci Alves de; Silva, Luzimar Campos daThe 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.Item An efficient system for in vitro propagation of Bouchea fluminensis (Vell.) Mold. (Verbenaceae)(Acta Botanica Brasilica, 2013-10-31) Resende, Cristiano Ferrara de; Braga, Virgínia Fernandes; Silva, Cristiane Jovelina da; Pereira, Paula da Fonseca; Ribeiro, Cleberson; Salimena, Fátima Regina Gonçalves; Peixoto, Paulo Henrique PereiraThis study aimed to establish and propagate in vitro plants of Bouchea fluminensis, a medicinal species known in Brazil as gervão-falso ("false verbena"), evaluating the influences of different growth regulators on in vitro multiplication and rooting stages, as well as examining ex vitro acclimatization of rooted plants. Explants were established on Murashige and Skoog medium at half strength of salts and vitamins without growth regulators. For multiplication, the explants were subjected to combinations of 6-benzyladenine (BA; 0, 2.5, 5.0 and 7.5 µM) and α-naphthalene-acetic acid (NAA; 0, 0.2, 0.4 and 0.6 µM). The medium found to induce the greatest number of shoot was that containing 5 µM of BA (NAA-free). For rooting, we evaluated three auxins (NAA, indole-3-acetic acid and indole-3-butyric acid; 0.1, 0.2, 0.3 and 0.4 µM), as well as a control. No differences were observed between the control and the other treatments. The auxin-free medium was deemed the most suitable, because it ensures the lowest cost in the micropropagation procedures. We obtained 100% survival of the acclimatized seedlings, and the plants showed normal vegetative and reproductive development, suggesting that the micropropagation did not alter the biological cycle of this species. The results show the importance and potential of micropropagation for biodiversity conservation of Bouchea fluminensis.Item Excesso de ferro sobre o crescimento e a composição mineral em Eugenia uniflora L.(Revista Ciência Agronômica, 2016-10) Jucoski, Gládis de Oliveira; Cambraia, José; Ribeiro, Cleberson; Oliveira, Juraci Alves deO ferro, embora micronutriente essencial, quando em excesso pode causar redução no crescimento e, consequentemente, na produtividade das plantas. O objetivo do presente trabalho foi avaliar a absorção, o acúmulo, a distribuição do Fe e suas consequências sobre o crescimento e composição mineral de plantas jovens de Eugenia uniflora L.. As plantas foram expostas às concentrações de Fe 0,045 (controle), 1,0 e 2,0 mM, aplicado na forma de FeEDTA, em solução nutritiva de Hoagland, pH 5,0, durante 15; 30 e 45 dias e, então, foram avaliados o número de folhas, a altura da parte aérea, o comprimento da raiz primária, a massa seca de raízes, caule e folhas e os teores de clorofila, carotenóides totais e minerais. Plantas expostas aos tratamentos com Fe 1,0 e 2,0 mM apresentaram aumento nos teores deste elemento em folhas, caule e raízes em relação às plantas-controle, especialmente aos 45 dias de exposição. Sob esta condição, as plantas exibiram sintomatologia típica de toxidez de Fe, caracterizada por bronzeamento foliar, escurecimento das raízes, redução no número de folhas, na altura da parte aérea, no comprimento da raiz principal, na produção de massa seca e nos teores de pigmentos cloroplastídicos. O excesso de Fe modificou a partição da biomassa e promoveu redução nos teores de P, Zn, Cu e Mn, especialmente nas raízes. Além de um efeito direto do Fe em excesso, a desordem nutricional resultante pode estar associada aos efeitos restritivos sobre o crescimento vegetativo inicial das plantas de Eugenia uniflora L..Item Nitric oxide attenuates oxidative stress induced by arsenic in lettuce (Lactuca sativa) leaves(Water, Air, & Soil Pollution, 2015-11) Silveira, Neidiquele M.; Canatto, Regiane A.; Siman, Luhan; Oliveira, Juraci A. de; Ribeiro, Cleberson; Cambraia, José; Farnese, FernandaLettuce plants were exposed to different toxic levels of arsenic (As) to induce an oxidative stress response, and the role of nitric oxide (NO) (provided as sodium nitroprusside (SNP)) as an attenuating agent of this stress condition was evaluated. Plants were treated with 50 μM of As with or without 100 μM SNP added to the nutrient solution. The hydrogen peroxide, superoxide anion, and malondialdehyde concentrations and enzymatic activities were measured. The increase in As concentration detected in the leaves was followed by a significant increase in H2O2 and malondialdehyde (MDA) concentrations. However, the presence of SPN promoted a reduction in the concentration of these oxidative agents and also reduced the translocation of As to the shoots. The enzymatic activities in the plants exposed to As were increased, which indicates the active participation of these enzymes in the reduction of oxidative stress induced by the metalloid. In the plants exposed to As and SNP, the enzymatic activities were not so high; this result was possibly related to the direct action of NO in scavenging the generated toxic metabolites and with the reduction in the translocation of the pollutant to the shoots. Lettuce and leaves of other vegetables are usually ingested, and this study shows an alternative to avoid human contamination with As.Item Role of glutathione in tolerance to arsenite in Salvinia molesta, an aquatic fern(Acta Botanica Brasilica, 2017-10) Silva, Adinan Alves da; Oliveira, Juraci Alves de; Campos, Fernanda Vidal de; Ribeiro, Cleberson; Farnese, Fernanda dos SantosIn many plant species, tolerance to toxic metals is highly dependent on glutathione, an essential metabolite for cellular detoxification. We evaluated the responses of glutathione metabolism to arsenite (AsIII) in Salvinia molesta, an aquatic fern that has unexplored phytoremediation potential. Plants were exposed to different AsIII concentrations in nutrient solution for 24 h. AsIII caused cell membrane damage to submerged leaves, indicating oxidative stress. There was an increase in the glutathione content and ϒ-glutamylcysteine synthetase enzyme activity in the submerged and floating leaves. The glutathione peroxidase and glutathione sulfotransferase enzymes also showed increased activity in both plant parts, whereas glutathione reductase only showed increased activity in the submerged leaves. These findings suggest an important role for glutathione in the protection of S. molesta against the toxic effects of AsIII, with more effective tolerance responses in the floating leaves.Item Rudgea viburnoides (Rubiaceae) overcomes the low soil fertility of the Brazilian Cerrado and hyperaccumulates aluminum in cell walls and chloroplasts(Plant and Soil, 2016-11) Malta, Priscila Gonçalves; Ribeiro, Cleberson; Campos, Naiara Viana; Azevedo, Aristéa Alves; Arcanjo-Silva, SamaraRudgea viburnoides (Cham.) Benth. is an aluminum (Al) hyperaccumulator species native to the Brazilian Cerrado and can be found on soils with different fertilities and Al availabilities. We studied the relationship between the chemical composition of different soils and the Al and nutrient concentrations in the species. Histolocalization of Al and its probable complexation with phenols were investigated. Aluminum in R. viburnoides roots, stems and leaves was detected through histochemical tests, confocal microscopy, and X-ray microanalysis. Phenol localization and concentration were determined, as was the plant nutritional status. Despite the low soil fertilities, nutrient concentrations in vegetative organs were high. Even with the differences in Al availability in the soils, R. viburnoides accumulated more than 10 g Al kg−1 DW on leaves. Pectocellulosic cell walls were preferential sites for Al deposition, but the metal also accumulated in suberized cell walls and chloroplasts. Aluminum localization coincided with Si distribution in cell walls and phenol distribution in the cytosol. Rudgea viburnoides has a great ability to absorb nutrients from poor soils and the hyperaccumulation of Al is independent of the metal availability in the soil. Aluminum accumulation on the phellem and chloroplasts was described for the first time in this species.Item Sistemas de defesa contra estresses oxidativos em dois cultivares de arroz (Oryza sativa L.) com tolerância diferencial ao alumínio(Universidade Federal de Viçosa, 2007-03-08) Ribeiro, Cleberson; Peixoto, Paulo Henrique Pereira; http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4780011D1; Cano, Marco Antonio Oliva; http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4787546T4; Cambraia, José; http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4783868U6; http://lattes.cnpq.br/5023387764069051; Silva, Marco Aurélio Pedron e; http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4790898P8; Oliveira, Juraci Alves de; http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4782512D8Os efeitos do alumínio (Al) sobre o crescimento e sobre os sistemas de defesa antioxidativos enzimáticos e não-enzimáticos envolvidos na eliminação dos intermediários reativos de oxigênio (ROIs) foram avaliados em dois cultivares de arroz: Fernandes (CNA-1158) e Maravilha (CNA-6843-1) com tolerância diferencial ao Al. As plantas, cultivadas em solução nutritiva, pH 4,0, foram tratadas com Al nas concentrações de 0 e 1 mM, durante dez dias. A exposição das plantas ao Al não afetou o crescimento no cultivar Fernandes, enquanto, no Maravilha, reduções significativas foram observadas nos parâmetros de crescimento avaliados. Nas raízes dos dois cultivares, a presença do Al aumentou a atividade das enzimas catalases (CATs), peroxidases (POXs), redutases da glutationa (GRs) e peroxidases da glutationa (GPXs). A atividade das superóxido dismutases (SODs) aumentou apenas nas raízes do cultivar Fernandes, não sendo modificada para o cultivar Maravilha. Independente do tratamento aplicado e da parte da planta analisada, a atividade das peroxidases do ascorbato (APXs) foi sempre maior no cultivar Fernandes. Nas folhas, o Al não alterou a atividade de nenhuma das enzimas no cultivar Fernandes, exceto a das POXs que sofreu redução. No cultivar Maravilha, por outro lado, as atividades das SODs, POXs e GPXs foram reduzidas na presença de Al. Dentre as enzimas estudadas, as SODs, APXs e GPXs nas raízes e as CATs, POXs, APXs nas folhas, exibiram resposta consistente com a tolerância diferencial ao Al apresentada pelos dois cultivares de arroz estudados. Nos dois cultivares, o teor de ascorbato (AA) aumentou nas folhas e o de desidroascorbato (DHA) reduziu nas raízes, em resposta ao tratamento com Al. O teor da forma reduzida (AA) foi muito mais elevado nas folhas, na qual a relação AA/DHA atingiu valores 30 vezes maiores que nas raízes. Nas raízes, o cultivar Fernandes apresentou menores teores de AA, porém maior atividade das APXs, enquanto no cultivar Maravilha foi observado o oposto, demonstrando o importante papel desse metabólito como substrato para a reação catalisada pelas APXs. A concentração de glutationa total, também, parece ser importante no sistema de defesa não enzimático, mas provavelmente seria necessário discriminar entre as suas formas, reduzida (GSH) e oxidada (GSSG), para entender seu papel como substrato das enzimas antioxidativas. De modo geral, as atividades dos sistemas de defesa antioxidativos enzimático e não-enzimático, tanto nas raízes como nas folhas dos dois cultivares, indicaram ter o cultivar Fernandes mecanismos de defesa mais eficientes no combate aos ROIs produzidos durante o tratamento com Al.