RESUMO Objetivou-se com este trabalho identificar a resposta antioxidante de variedades de cana-de-açúcar sob condições adversas. Mudas de seis genótipos, procedentes da micropropagação, foram submetidas à avaliação da ação isolada e combinada do estresse hídrico e salino. A pesquisa foi realizada em maio de 2012, em casa de vegetação instalada na Universidade Federal Rural de Pernambuco (UFRPE). O delineamento experimental foi o inteiramente casualizado, com tratamentos dispostos em esquema fatorial 6 x 4 (seis variedades e quatro condições de cultivo) com quatro repetições. Avaliou-se a peroxidação lipídica, o teor de peróxido de hidrogênio (H2O2), o teor relativo de água (TRA) e ação do sistema de defesa antioxidativo mediante a atividade das enzimas superóxido dismutase (SOD), ascorbato peroxidase (APX) e catalase (CAT). As variedades de cana-de-açúcar exibiram maior peroxidação lipídica e/ou maior concentração de peróxido de hidrogênio quando submetidas ao estresse combinado. As enzimas apresentaram respostas diferenciadas de acordo com a variedade de cana-de-açúcar e a intensidade de estresse nos tratamentos isolados, no entanto, podem ter sido inativadas perante o estresse combinado, o que indica que a resposta do estresse combinado é diferente da soma das respostas dos fatores abióticos aplicados isoladamente. Os elevados teores de malondialdeído (MDA) associados ao baixo TRA podem ser indicadores efetivos de sensibilidade a estresse múltiplo em variedades de cana-de-açúcar. As variedades RB99395 e RB867515 apresentaram-se mais eficientes quanto a resposta ao estresse ambiental e a manutenção de conteúdo hídrico celular nos estresses isolados.

ABSTRACT This study aimed to identify the antioxidant responses of sugarcane (Saccharum spp.) varieties subjected to water and saline stress. Sugarcane seedlings of six different varieties obtained through micropropagation were subjected to either water or saline stress, or a combination of water + saline stress. The study was carried out in May 2012, in a greenhouse at the Universidade Federal Rural de Pernambuco (UFRPE). The experimental design was randomized, with treatments arranged in a 6 × 4 factorial scheme (six varieties and four treatments), and four replicates. Lipid peroxidation, hydrogen peroxide (H2O2) concentrations, and relative water content (RWC) were evaluated. Furthermore, we evaluated the plants’ antioxidative defense systems by measuring the activities of superoxide dismutase (SOD), ascorbate peroxidase (APX), and catalase (CAT). The sugarcane varieties had higher lipid peroxidation and/or higher H2O2 concentrations when subjected to the combined water + saline stress. The antioxidant enzymes responded to the water and saline stress treatments differently depending on the sugarcane variety. However, under combined saline + water stress conditions, the enzymes may have become inactivated, which indicates that the response to the combined water + saline stress was different from the sum of the responses to only water stress or only saline stress. High concentrations of malondialdehyde (MDA) associated with low RWC may be an effective indicator of multiple stress sensitivity in sugarcane varieties. The RB99395 and RB867515 sugarcane varieties responded more efficiently to environmental stress, and maintained their cell water content when subjected to either water or saline stress.

Abstract The objective of this study was to investigate and characterize cowpea (Vigna unguiculata) genotypes for total grain protein content, storage protein fractions (globulin, albumin, prolamin, basic and acid glutelins), and phytate and minerals contents. Eighteen cowpea genotypes were selected. Total grain protein content varied from 21.4% to 29.2%, for BRS Marataoã and Paulistinha genotypes, respectively. The variation in the concentration of each protein fraction was significant (P<0.05) only for glutelins (basic and acid). The genotypes studied exhibited great similarity in the PAGE electrophoretic profile of the grain protein fractions and also in the mineral content. BRS Paraguaçu genotype exhibited higher Zn content than thegenotypes that have been previously recommended for this characteristic. The lowest phytate grain content was observed in four of the 18 genotypes studied, which also exhibited high protein contents. Although the results did not converge to the selection of a few genotypes, some specific differences were detected that which may be further explored. Considering total grain protein, mineral and phytate contents, the genotype Paulistinha revealed a better balance unveiling high grain total protein content, low grain phytate content and more homogeneous mineral composition.

In the present work, dried leaves of Saururus cernuus (Saururaceae) were subjected to extraction using an ionic liquid (1-butyl-3-methylimidazolium bromide - BMImBr) in the microwave assisted extraction (MAE). The obtained extract was partitioned using n-hexane and cytotoxicity activity of this organic phase against murine melanoma cell line (B16F10-Nex2) was evaluated in vitro. Since this extract displayed activity (100% of cell death at 200 µg mL-1) it was subjected to a bioactivity-guided fractionation to afford four related neolignans: threo-austrobailignan-5 (1), threo-austrobailignan-6 (2), threo-dihydroguaiaretic acid (3) and saucernetin (4). Their chemical structures were established based on NMR and MS spectral analysis. Among the isolated neolignans, compound 2 exhibited the highest cytotoxic activity against HeLa (human cervical melanoma) cells with IC50 of 28.3 ± 3.9 µg mL-1 (86 ± 12 µmol L-1). Furthermore, compounds 2 and 3 exhibited the highest cytotoxic activity against A2058 (human melanoma) cells with IC50 of 44.3 ± 4.2 µg mL-1 (135 ± 13 µmol L-1) and 41.5 ± 7.5 µg mL-1 (126 ± 23 µmol L-1), respectively, similar to positive control cisplatin (IC50 = 43.2 ± 3.2 µg mL-1 or 144 ± 11 µmol L-1). Otherwise, compound 4 was inactive (IC50 > 100 µg mL-1 or > 300 µmol L-1). The obtained results provide important data for the selection of bioactive neolignans with promising cytotoxic potential using a simple and fast method employing a green solvent as 1-butyl-3-methylimidazolium bromide (BMImBr).

ABSTRACT In this study, we are presenting recommendations to the best agricultural use as well as for plant breeding of three millet cultivars namely ENA1 and ENA2, which have African origin, and BRS1501 originally from India. These cultivars were evaluated for growth, yield and grain quality traits. The morphological traits evaluated in this study indicated that the African genotypes ENA1 and ENA2 are better than the Indian genotype BRS1501 for no-till farming or to produce forage with 15% of crude protein at flowering and at harvest to produce stover (around 7% of crude protein content) for livestock feeding. The BRS1501 cultivar exhibited the highest values for total crude protein, albumins and prolamins, phytate and mineral contents in grains. ENA1 and ENA2 exhibited the highest values of globulin and glutelin contents. The electrophoretic patterns for storage proteins were similar across the three millets cultivars, except for a higher intensity of two glutelin bands with 21 and 24 kDa in BRS1501. Together, the results allow us to recommend BRS1501 for grain production and ENA1 and ENA2 for biomass production.

ABSTRACT Both the scientific community and society have shown interest in improving the content of amino acids, carbohydrates and mineral nutrients in maize because it represents an important staple food in many developing countries. Earlier studies demonstrated that the treatment of seeds using ascorbic acid (AsA-seed priming) enhanced soluble carbohydrates, proteins and soluble amino acids for other species. AsA seed priming in maize showed the potential for reducing abiotic stresses. The effects on grain quality have not been previously demonstrated. This study investigated the impacts of AsA seed priming on maize kernel quality of seeds produced by the plants generated from the primed seeds, based on the amino acid profile and carbohydrate and mineral nutrient contents. AsA seed priming improved the maize kernel quality with respect to the ascorbate content, boron allocation, total carbohydrate content and increased soluble amino acid levels, including serine, tyrosine, alanine, valine, glutamate, arginine, proline, aspartate, lysine and isoleucine, whereas soluble methionine was decreased. Therefore, AsA seed priming can represent a potential technique for improving maize grain quality.

A proteômica é uma área incrível da ciência cujas apli cações vêm avançando para diversos propósitos. Um aspecto crucial para se alcançar boa resolução proteômica é o estabelecimento de uma metodologia que resulte na melhor qualidade e ampla representação das proteínas totais. Outro aspecto importante é que em muitos estudos quantidades limitadas de tecido e de proteínas totais no tecido a ser estudado são encontradas, tornando difícil a análise. Com o objetivo de testar diferentes parâmetros, combinações usando mínimas quantidades de tecido com 4 protocolos para extração protéica de folhas e raízes de tomateiro (Solanum lycopersicumL.) foram avaliadas com atenção especial para a capacidade de remover interferentes e atingir resolução adequada em eletroforese bidimensional, bem como rendimento satisfatório de proteínas. A avaliação dos protocolos de extração revelou grandes diferenças na quantidade de proteínas obtidas por cada um. O protocolo TCA/acetona mostrou ser o protocolo mais eficiente, que permitiu a detecção de 211 "spots" para folhas e 336 para raízes utilizando 500 µg de proteína de folhas e 800 µg de proteínas de raiz por gel.

Proteomics is an outstanding area in science whose increasing application has advanced to distinct purposes. A crucial aspect to achieve a good proteome resolution is the establishment of a methodology that results in the best quality and wide range representation of total proteins. Another important aspect is that in many studies, limited amounts of tissue and total protein in the tissue to be studied are found, making difficult the analysis. In order to test different parameters, combinations using minimum amount of tissue with 4 protocols for protein extraction from tomato (Solanum lycopersicum L.) leaves and roots were evaluated with special attention to their capacity for removing interferents and achieving suitable resolution in bidimensional gel electrophoresis, as well as satisfactory protein yield. Evaluation of the extraction protocols revealed large protein yield differences obtained for each one. TCA/acetone was shown to be the most efficient protocol, which allowed detection of 211 spots for leaves and 336 for roots using 500 µg of leaf protein and 800 µg of root protein per gel.

A peroxidação lipídica e alongamento de raiz de Eucalyptus grandis × Eucalyptus camaldulensis foram estudados em resposta às condições de estresse com alumínio (Al), um metal conhecido por limitar a produtividade agrícola em solos ácidos, principalmente devido à redução do crescimento radicular. No Brasil, o híbrido Grancam 1277 (E. grandis × E. camaldulensis) tem sido plantado na região do "Cerrado", local com grande ocorrência de solos ácidos. O presente estudo mostra que o híbrido teve redução de crescimento das raízes e aumento dos níveis de peroxidação lipídica, após 24h de tratamento com 100 µM de Al. No entanto, um possível processo de tolerância começou após este período, uma vez que foram observadas redução na peroxidação lipídica e recuperação de alongamento da raiz após 48h de exposição Al. A compreensão da dinâmica temporal de tolerância ao Al pode ser útil principalmente para selecionar genótipos mais tolerantes e também para identificar genes de interesse para aplicação em bioengenharia.

Lipid peroxidation and root elongation of Eucalyptus grandis × Eucalyptus camaldulensis were studied under stress conditions in response to aluminum (Al), a metal known to limit agricultural productivity in acidic soils primarily due to reduced root elongation. In Brazil, the Grancam 1277 hybrid (E. grandis × E. camaldulensis) has been planted in the "Cerrado", a region of the country with a wide occurrence of acidic soils. The present study demonstrated that the hybrid exhibited root growth reduction and increased levels of lipid peroxidation after 24h of treatment with 100 µM of Al, which was followed by a reduction in lipid peroxidation levels and the recovery of root elongation after 48h of Al exposure, suggesting a rapid response to the early stressful conditions induced by Al. The understanding of the temporal dynamics of Al tolerance may be useful for selecting more tolerant genotypes and for identifying genes of interest for applications in bioengineering.

Neste trabalho, analisamos as respostas bioquímicas inerentes ao sistema antioxidante, assim como propriedades morfológicas e anatômicas de mutantes fotomorfogenéticos e hormonais de tomateiro. Comparados ao não mutante Micro-Tom (MT), observamos que o conteúdo de malondialdeído (MDA) aumentou nos mutantes diageotropica (dgt) e lutescent (l), enquanto os maiores níveis de H2O2 foram encontrados nos mutantes high pigment 1 (hp1) e aurea (au). Análises de enzimas antioxidantes mostraram que todos os mutantes reduziram a atividade de catalase (CAT) quando comparado a MT. A guaiacol peroxidase (GPOX) aumentou em ambos os mutantes sitiens (sit) e notabilis (not), enquanto no mutante not houve aumento da atividade de ascorbato peroxidase (APX). Baseado nas análises de PAGE, as isoformas III, IV, V e VI de glutationa redutase (GR) aumentaram nas folhas do mutante l, enquanto a isoforma III de superóxido dismutase (SOD) reduziu nas folhas dos mutantes sit, epi, Never ripe (Nr) e green flesh (gf). Análises por microscopia revelaram que os mutantes hp1 e au tiveram um aumento nos espaços intercelulares de folhas, enquanto o mutante sit mostrou um decréscimo. Micrografias eletrônicas revelaram que a superfície foliar dos mutantes au e hp1 apresenta reduzida distribuição de tricomas. A caracterização destes mutantes é essencial para que possam ser utilizados em estudos de desenvolvimento e ecofisiologia, tais como estresses abióticos e bióticos no metabolismo oxidativo.

Biochemical responses inherent to antioxidant systems as well morphological and anatomical properties of photomorphogenic, hormonal and developmental tomato mutants were investigated. Compared to the non-mutant Micro-Tom (MT), we observed that the malondialdehyde (MDA) content was enhanced in the diageotropica (dgt) and lutescent (l) mutants, whilst the highest levels of hydrogen peroxide (H2O2) were observed in high pigment 1 (hp1) and aurea (au) mutants. The analyses of antioxidant enzymes revealed that all mutants exhibited reduced catalase (CAT) activity when compared to MT. Guaiacol peroxidase (GPOX) was enhanced in both sitiens (sit) and notabilis (not) mutants, whereas in not mutant there was an increase in ascorbate peroxidase (APX). Based on PAGE analysis, the activities of glutathione reductase (GR) isoforms III, IV, V and VI were increased in l leaves, while the activity of superoxide dismutase (SOD) isoform III was reduced in leaves of sit, epi, Never ripe (Nr) and green flesh (gf) mutants. Microscopic analyses revealed that hp1 and au showed an increase in leaf intercellular spaces, whereas sit exhibited a decrease. The au and hp1 mutants also exhibited a decreased in the number of leaf trichomes. The characterization of these mutants is essential for their future use in plant development and ecophysiology studies, such as abiotic and biotic stresses on the oxidative metabolism.

As adaptações ecofisiológicas de aguapé (hiperacumuladora de metais) sob condições ambientais adversas ainda não estão totalmente elucidadas. Este trabalho avaliou pigmentos fotossintéticos e fluorescência da clorofila a em aguapé coletados em diferentes regiões (alto, médio e baixo) do rio Paraíba do Sul (RPS) e no rio Imbé, Brasil. Adicionalmente Mn, Fe, Zn, Cu, Pb e Ni foram quantificados na parte área e raízes. Ferro e Mn foram os elementos detectados em maiores concentrações em todas as amostras. Zinco e Cu foram detectados em maiores concentrações nas raízes de plantas do médio e alto RPS, locais de grande atividade antropogênica. As mais elevadas concentrações de quase todos os metais foram verificadas nas plantas do médio RPS. Com relação às adaptações fotossintéticas, plantas do médio e alto RPS mostraram as maiores concentrações de pigmentos fotossintéticos, assim como os mais elevados valores de Fv/Fm e Fv/F0. As avaliações fotossintéticas destas plantas sugerem que elas não estejam sob condições de estresse, mesmo nas regiões de grande atividade antropogênica. Correlação positiva foi observada entre quenching não-fotoquímico e carotenóides, sugerindo que a dissipação de energia na forma de calor seja uma estratégia para manter o funcionamento fotossintético fotoquímico estável, confirmado pelos valores de Fv/Fm. É possível que esta estratégia adaptativa seja eficiente como resposta de longo prazo, uma vez que na literatura esta não parece ser a via preferencial de adaptação a estresses de curto prazo, avaliados sob condições controladas.

The ecophysiological adaptations of water hyacinth (metal hyperaccumulator) under adverse environmental conditions are not yet clearly elucidated. This work evaluated photosynthetic pigments and chlorophyll a fluorescence in water hyacinth plants sampled in different regions (upper, middle and lower) of Paraíba do Sul river (PSR) and Imbé river, Brazil. Additionally Mn, Fe, Zn, Cu, Cr, Pb and Ni were quantified on the shoots and roots. Roots were the preferential site of metal accumulation. Cromium was not detected in the shoots. Iron and Mn were the elements detected in higher concentrations in all samples. Zinc and Cu were detected in higher concentrations in roots of plants from middle and upper PSR, sites of higher anthropogenic activity. The highest concentrations of almost all metals were verified in plants from middle PSR. Regarding photosynthetic adaptations, plants from middle and upper PSR showed the highest concentrations of photosynthetic pigments, as well as the highest values of Fv/Fm and Fv/F0. The photosynthetic performance of plants suggests that they are not under stress conditions, even in regions of higher anthropogenic activity. Positive correlation was observed among nonphotochemical quenching and carotenoids, suggesting dissipation of energy in the form of heat as an strategy to keep the stable photochemical photosynthesis functioning, confirmed by values of the Fv/Fm. It is possible that such adaptative strategy is efficient as a long term response to stress factors, once literature data suggest that this is not a preferential path for photosynthesis in stress circumstances under short time controlled conditions.

O gene Opaco-2 (O2), expresso especificamente no grão de milho, transcreve para um fator de transcrição da família "leucine-zipper". Mutantes o2 apresentam grãos opacos, redução na quantidade de zeínas e aumento na proporção de lisina e triptofano. Genes cuja expressão é controlada diretamente pelo O2 são conhecidos (alfa-zeínas de 22 kDa, beta-zeínas de 14 kDa, b-32 e cyPpdk1). Nesta revisão, nós apresentamos resultados da caracterização genética de genes relacionados com o O2, através de abordagens de transcritoma, proteoma e de atividades enzimáticas da via metabólica do aspartato e da degradação da lisina. O polimorfismo do locus O2 foi avaliado utilisando-se a técnica de RFLP em 51 linhagens de milho. A maioria dos polimorfismos foi observada nas regiões não codificadoras da proteína. Análises de correlação foram realizadas entre os polimorfismos de RFLP e (i) quantidade de RNAm do O2, cyPpdk, Lor/Sdh e Ahas (ii) quantidade de isoformas de zeínas e (iii) atividade da enzima SDH. Sítios polimórficos foram correlacionados com a quantidade de RNAm do próprio O2, do gene Lor/Sdh e com a quantidade de duas isoformas de a-zeinas de 19 kDa. Nossos resultados indicam a presença de relações entre o polimorfismo do locus O2 e o nível de expressão de genes sob o seu controle. A utilização de um maior número de linhagens e o uso de dados de seqüência do O2 permitirá uma análise precisa da conseqüência do polimorfismo deste fator de transcrição sobre o controle do seu próprio nível de expressão e dos genes por ele controlados.

The Opaque-2 (O2) gene encodes a transcriptional activator specifically expressed for grain development of maize. o2 mutants have an opaque and chalky kernel, with a decrease in zein storage protein content, and an increase in the proportions of lysine and tryptophan. In this review, we present recent results investigating genetic properties of the O2 network, using transcriptome and proteome approaches, associated with measurements of activities of enzymes of the aspartate pathway and lysine degradation. The structural polymorphism at the O2 locus was investigated by RFLP in a collection of 51 maize inbred lines. Most polymorphic sites were found outside the coding regions. We then searched for relationships between RFLP polymorphism and (i) mRNA abundance of O2 and of known or suspected target genes, (ii) activity of SDH and (iii) amount of zein isoforms. Polymorphic restriction sites in the 5' upstream regions of the O2 gene were found associated with O2 mRNA abundance (three sites) and the amount of two 19 kDa alpha-zein isoforms (two sites). One restriction site on the 3' side of the O2 gene was found associated with Lor/Sdh mRNA abundance. Our results indicate relationships between polymorphism at the O2 locus and the expression of some of its target genes. Evidence of these associations has to be confirmed on larger samples, and the analysis of the O2 gene sequence should allow more precise testing of the actual involvement of O2 polymorphism in its own transcriptional expression, and in the expression of its target genes.