Determination of macro- and micronutrients in plant leaves by high-resolution continuum source flame atomic absorption spectrometry combining instrumental and sample preparation strategies

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

The plant energy-dissipating mitochondrial systems: Depicting the genomic structure and the expression profiles of the gene families of uncoupling protein and alternative oxidase in monocots and dicots

The simultaneous existence of alternative oxidases and uncoupling proteins in plants has raised the question as to why plants need two energy-dissipating systems with apparently similar physiological functions. A probably complete plant uncoupling protein gene family is described and the expression profiles of this family compared with the multigene family of alternative oxidases in Arabidopsis thaliana and sugarcane (Saccharum sp.) employed as dicot and monocot models, respectively. In total, six uncoupling protein genes, AtPUMP1-6, were recognized within the Arabidopsis genome and five (SsPUMP1-5) in a sugarcane EST database. The recombinant AtPUMP5 protein displayed similar biochemical properties as AtPUMP1. Sugarcane possessed four Arabidopsis AOx1-type orthologues (SsAOx1a-1d); no sugarcane orthologue corresponding to Arabidopsis AOx2-type genes was identified. Phylogenetic and expression analyses suggested that AtAOx1d does not belong to the AOx1-type family but forms a new (AOx3-type) family. Tissue-enriched expression profiling revealed that uncoupling protein genes were expressed more ubiquitously than the alternative oxidase genes. Distinct expression patterns among gene family members were observed between monocots and dicots and during chilling stress. These findings suggest that the members of each energy-dissipating system are subject to different cell or tissue/organ transcriptional regulation. As a result, plants may respond more flexibly to adverse biotic and abiotic conditions, in which oxidative stress is involved. © The Author [2006]. Published by Oxford University Press [on behalf of the Society for Experimental Biology]. All rights reserved.

Carbohydrate Composition and Water-Stable Aggregation of an Oxisol as Affected by Crop Sequence under No-Till

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Acúmulo e distribuição de boro em cultivares de algodão

O algodoeiro é responsivo à adubação boratada, porém há poucas e controversas observações a respeito da mobilidade do nutriente dentro da planta. Este trabalho teve por objetivo avaliar a translocação e o acúmulo de boro (B) em cultivares de algodão. Quatro cultivares de algodão (FM 993, FM 910, FMT 701 e FMT 523) foram cultivadas em duas concentrações de B (0 e 4,5 mmol L-1) na solução nutritiva, em vasos com capacidade de 8 L preenchidos com areia lavada, por seis semanas. Os vasos receberam solução nutritiva uma vez por semana. em seguida, as plantas foram amostradas durante quatro semanas consecutivas, determinando-se a matéria seca e acúmulo total de B nas raízes e na parte aérea, a distribuição e acúmulo de B nos tecidos da planta e o teor e acúmulo de B nas folhas em diferentes nós da haste principal. O delineamento experimental foi em blocos ao acaso, em esquema fatorial 4 x 2, com parcelas subdivididas no tempo (semanas de amostragem), com quatro repetições. As cultivares FM 993 e FMT 523 alcançaram maior produção de matéria seca e acúmulo de B na raiz. Plantas cultivadas sem deficiência de B acumulam mais de 50 % do B nas folhas, ao passo que, em plantas sem aplicação do micronutriente, o B se encontra em maior proporção nas raízes e no caule. Não há evidência de translocação de B em diferentes cultivares de algodão, mesmo quando cultivadas em deficiência desse nutriente.

The effect of pH on horseradish peroxidase-catalyzed oxidation of melatonin: production of N-1-acetyl-N-2-formyl-5-methoxykynuramine versus radical-mediated degradation

There is a g-rowing body of evidence that melatonin and its oxidation product, N-1-acetyl-N-2-formyl-5-methoxykynuramine (AFMK), have anti-inflammatory properties. From a nutritional point of view, the discovery of melatonin in plant tissues emphasizes the importance of its relationship with plant peroxidases. Here we found that the pH of the reaction mixture has a profound influence in the reaction rate and products distribution when melatonin is oxidized by the plant enzyme horseradish peroxidase. At pH 5.5. 1 mm of melatonin was almost completely oxidized within 2 min, whereas only about 3% was consumed at pH 7.4. However, the relative yield of AFMK was higher in physiological pH. Radical-mediated oxidation products, including 2-hydroxymelatonin a dimer of, 2-hydroxymelatonin and O-demethylated dimer of melatonin account for the fast consumption of melatonin at pH 5.5. The higher production of AFMK at pH 7.4 was explained by the involvement of compound III of peroxidases as evidenced by spectral studies. on the other hand, the fast oxidative degradation at pH 5.5 was explained by the classic peroxidase cycle.

Fast sequential multi-element determination of Ca, Mg, K, Cu, Fe, Mn and Zn for foliar diagnosis using high-resolution continuum source flame atomic absorption spectrometry: Feasibility of secondary lines, side pixel registration and least-squares background correction

The fast sequential multi-element determination of Ca, Mg, K, Cu, Fe, Mn and Zn in plant tissues by high-resolution continuum source flame atomic absorption spectrometry is proposed. For this, the main lines for Cu (324.754 nm), Fe (248.327 nm), Mn (279.482 nm) and Zn (213.857 nm) were selected, and the secondary lines for Ca (239.856 nm), Mg (202.582 nm) and K (404.414 nm) were evaluated. The side pixel registration approach was studied to reduce sensitivity and extend the linear working range for Mg by measuring at wings (202.576 nm; 202.577 nm; 202.578 nm; 202.580 nm: 202.585 nm; 202.586 nm: 202.587 nm; 202.588 nm) of the secondary line. The interference caused by NO bands on Zn at 213.857 nm was removed using the least-squares background correction. Using the main lines for Cu, Fe, Mn and Zn, secondary lines for Ca and K, and line wing at 202.588 nm for Mg, and 5 mL min(-1) sample flow-rate, calibration curves in the 0.1-0.5 mg L-1 Cu, 0.5-4.0 mg L-1 Fe, 0.5-4.0 mg L-1 Mn, 0.2-1.0 mg L-1 Zn, 10.0-100.0 mg L-1 Ca, 5.0-40.0 mg L-1 Mg and 50.0-250.0 mg L-1 K ranges were consistently obtained. Accuracy and precision were evaluated after analysis of five plant standard reference materials. Results were in agreement at a 95% confidence level (paired t-test) with certified values. The proposed method was applied to digests of sugar-cane leaves and results were close to those obtained by line-source flame atomic absorption spectrometry. Recoveries of Ca, Mg, K, Cu, Fe, Mn and Zn in the 89-103%, 84-107%, 87-103%, 85-105%, 92-106%, 91-114%, 96-114% intervals, respectively, were obtained. The limits of detection were 0.6 mg L-1 Ca, 0.4 mg L-1 Mg, 0.4 mg L-1 K, 7.7 mu g L-1 Cu, 7.7 mu g L-1 Fe, 1.5 mu g L-1 Mn and 5.9 mu g L-1 Zn. (C) 2009 Elsevier B.V. All rights reserved.

Fracionamento de manganês acumulado nos tecidos de mudas de caramboleira

A caramboleira (Averrhoa carambola) tende a acumular grande quantidade de Mn nos seus tecidos, mas são escassas as informações sobre a forma como o excesso desse nutriente permanece na planta. Com o objetivo de contribuir para o esclarecimento desse aspecto, neste trabalho foram cultivadas mudas de caramboleira em solução nutritiva de Furlani (1999) com concentrações crescentes de Mn (0; 0,5; 25; e 50 mg L-1 de Mn), e aos 30, 60, 90 e 120 dias os tecidos de caule, folha e raiz foram colhidos e submetidos a uma sequência de soluções extratoras: água, DTPA (ácido dietilenotriaminopentacético) e HCl (ácido clorídrico) 1 mol L-1, com posterior determinação da quantidade de Mn nos extratos e nos tecidos remanescentes. Foram encontrados teores crescentes de Mn nos extratos de água, DTPA e HCl 1 mol L-1. A maior parte (cerca de 50 %) do Mn permaneceu no tecido vegetal, mesmo após subsequentes extrações com as diferentes soluções extratoras, indicando que essa parte está fortemente ligada aos tecidos. Os maiores teores de Mn foram encontrados nas raízes e, os menores, no caule e nas folhas, sugerindo que a caramboleira tende a fixar o Mn nas raízes, possivelmente como mecanismo para limitar o transporte para a parte área, evitando a intoxicação da planta.

Faixas normais de nutrientes pelos métodos ChM, DRIS e CND e nível crítico pelo método de distribuição normal reduzida para laranjeira-pera

A utilização de métodos de diagnose nutricional para definição de teores ótimos e níveis críticos de nutrientes em tecidos vegetais tem se demonstrado promissora, desde que se conheçam suas limitações. Este trabalho teve como objetivo determinar as faixas normais de nutrientes para a cultura da laranjeira-pera em uma população, utilizando os métodos Chance Matemática (ChM), Sistema Integrado de Diagnose e Recomendação (DRIS) e Diagnose da Composição Nutricional (CND), além do Nível Crítico, pelo método de distribuição normal reduzida. O trabalho foi realizado no município de Bebedouro-SP, na Estação Experimental de Citricultura de Bebedouro. Utilizaram-se como base de dados teores totais de nutrientes de 50 amostras foliares e a produtividade da laranjeira-pera, oriundas de um experimento cujo fator de avaliação foram doses de calcário aplicadas superficialmente. Para o N, maior valor de ChM foi obtido pela classe 2 (23,6 a 24,7 g kg-1), com valores semelhantes aos obtidos pelo DRIS (22,1 a 24,0 g kg-1) e CND (22,1 a 23,9 g kg-1). Os valores inferiores dessas faixas normais concordam com o do nível crítico alcançado (22,7 g kg-1), sendo este muito próximo do proposto pela literatura. Para os nutrientes P, K, Mg, Zn e B, as faixas normais e os níveis críticos não se assemelharam aos descritos na literatura. em relação aos nutrientes Ca, Fe, Mn e Cu, seus valores de faixa normal e nível crítico aproximaram-se dos recomendados, possivelmente devido à maior variação em seus teores. A utilização dos métodos propostos, em uma população, foi mais adequada quando houve maior variação nos teores dos nutrientes, além de possibilitar menor amplitude aos valores de faixas normais, quando comparados aos da faixa de terras suficientes encontrados na literatura.

Tolerância à salinidade em feijão (Phaseolus vulgaris L)

Uma das aplicações das técnicas da cultura de tecidos no melhoramento é a identificação de linhas de células que apresentam tolerância à salinidade. Vários autores obtiveram linhas de células tolerantes ao estresse salino; e estudo de mecanismos bioquímicos da tolerância a sais em plantas tem demonstrado altas correlações entre estes e o acúmulo de macromoléculas em tecido de plantas superiores. Para verificar essas correlações em feijão (Phaseolus vulgaris cv IAC carioca), calos oriundos de eixos embrionários foram cultivados em meio sólido, suplementado com NaCl nas concentrações de 0 a 60 mM. Após 13 dias de incubação, os calos foram coletados e analisados quanto ao crescimento relativo, teor de proteínas, teor de prolina e atividade da peroxidase. Os parâmetros analisados mostraram decréscimo no crescimento relativo e no de proteínas em resposta ao NaCl. Paralelamente, observou-se aumento significativo no conteúdo de prolina e atividade da enzima peroxidase.

Effect of boron and zinc fertilization on white oats grown in soil with average content of these nutrients

The objective of this study was to evaluate the effect of fertilization with zinc or boron on the growth and dry matter production, nutritional value and accumulation of nutrients in white oats. The study comprised two experiments conducted in glasshouses, the first consisting of the application of four doses of zinc (0, 0.2, 0.4 and 0.6 mg/dm³) in the form of zinc sulphate (20% Zn), and the second consisting of the application of four doses of boron (0, 0.2, 0.4 and 0.6 mg/dm³) in the form of Borax (11% B). The experimental design in each case was a randomized block design, with five replicates. Fertilization with zinc and boron increased the growth of white oats, but had no significant effect on the nutritional value of the forage. Higher levels of absorption and accumulation of nutrients in plant tissues were observed following the application of boron and zinc at rates of up to 0.60 mg/dm³ of soil.

Produção de composto orgânico a partir de residuos de plantas medicinais

Extraction of active principles from medicinal plants generates wastes usually discharged. This process does not extract minerals from plant tissues, which may be considered of great value in organic composting. Wastes of 81 medicinal plant species processed at Centroflora/Anidro industry were analyzed for macro and micronutrients, and according to the results, three different ways to elaborate the compost were proposed: a) wastes of medicinal plants with high content of a specific mineral, aiming to fertilize soils where would be cultivated species that need high quantities of that mineral; b) wastes with mineral contents in order to support specific failures on mineral content of a soil; c) wastes of the eleven richest species in each nutrient that should elevate all mineral contents in soil.

The effect of pH on horseradish peroxidase-catalyzed oxidation of melatonin: Production of N1-acetyl-N2-formyl-5- methoxykynuramine versus radical-mediated degradation

There is a growing body of evidence that melatonin and its oxidation product, N1-acetyl-N2-formyl-5-methoxykynuramine (AFMK), have anti-inflammatory properties. From a nutritional point of view, the discovery of melatonin in plant tissues emphasizes the importance of its relationship with plant peroxidases. Here we found that the pH of the reaction mixture has a profound influence in the reaction rate and products distribution when melatonin is oxidized by the plant enzyme horseradish peroxidase. At pH 5.5, 1 mm of melatonin was almost completely oxidized within 2 min, whereas only about 3% was consumed at pH 7.4. However, the relative yield of AFMK was higher in physiological pH. Radical-mediated oxidation products, including 2-hydroxymelatonin, a dimer of 2-hydroxymelatonin and O-demethylated dimer of melatonin account for the fast consumption of melatonin at pH 5.5. The higher production of AFMK at pH 7.4 was explained by the involvement of compound III of peroxidases as evidenced by spectral studies. On the other hand, the fast oxidative degradation at pH 5.5 was explained by the classic peroxidase cycle. © 2007 The Authors.

Pectin and pectinases: Production, characterization and industrial application of microbial pectinolytic enzymes

Pectinases are a big group of enzymes that break down pectic polysaccharides of plant tissues into simpler molecules like galacturonic acids. It has long been used to increase yields and clarity of fruit juices. Since pectic substances are a very complex macromolecule group, various pectinolytic enzymes are required to degrade it completely. These enzymes present differences in their cleavage mode and specificity being basically classified into two main groups that act on pectin smooth regions or on pectin hairy regions. Pectinases are one of the most widely distributed enzymes in bacteria, fungi and plants. This review describes the pectinolytic enzymes and their substrates, the microbial pectinase production and characterization, and the industrial application of these enzymes. © Pedrolli et al.; Licensee Bentham Open.

Evaluation of sample preparation procedures and krypton as an interference standard probe for arsenic speciation by HPLC-ICP-QMS

Different conditions of extraction using water, a methanol-water mixture and nitric acid solutions were evaluated for speciation of As(iii), As(v), DMA and MMA in plant samples that previously received As(v) after being sown and emergence was investigated. Microwave-assisted extraction (MAE) using diluted nitric acid solutions was also performed for arsenic extraction from chicken feed samples. The separation and determination of arsenic species were performed using HPLC-ICP-MS. The interference standard method (IFS) using 83Kr+ as the IFS probe was employed to minimize spectral interferences caused by polyatomic species, such as 40Ar 35Cl+. The extraction procedures tested presented adequate extraction efficiencies (90%), and the four arsenic species evaluated were found in plant samples. Extractions with diluted nitric acid solution at 90 °C were the most efficient strategy, with quantitative recoveries for all four As species in plant tissues. On the other hand, the methanol-water mixture was the solvent with the lowest extraction efficiency (50-60%). For chicken feed samples, MAE at 100 °C for 30 min resulted in an extraction efficiency of 97% and only As(v) was found, without any species interconversion. The IFS method contributed to improving precision and limits of detection and quantification for all tested extraction procedures. Significant improvements on accuracy were obtained by applying the IFS method and recoveries between 77 and 94%, and 82 and 93% were obtained for plant extracts and chicken feed samples, respectively. This journal is © 2013 The Royal Society of Chemistry.

Prospecção de enzimas de degradação de materials vegetal em fungos endofíticos

Pós-graduação em Biotecnologia - IQ