Efeito do estresse hídrico e salino na germinação e vigor de sementes de maxixe

The objective of this work was to evaluate the germination and vigor of two lots of gherkin seeds in different osmotic potentials using polyethylene glycol (PEG 6000) and sodium chloride (NaCl). Seeds of the 'North' cultivar were sown in germitest paper moistened in solutions of polyethylene glycol and sodium chloride in osmotic potentials of 0, -0.3, -0.6 and -0.9MPa. The experimental design was completely randomized in a factorial 2x4 design (solutions x osmotic potential), with four replications. Seed vigor was evaluated by the germination test, first germination count, length and dry weight of root and hypocotyl. It is concluded that the reduction in osmotic potential from -0.3MPa induced by PEG 6000 and NaCl reduces the germination and vigor of gherkin's seeds. The osmotic stress induced by PEG 6000 produced more severe effects than saline in physiological quality of gherkin's seeds.

Estresse hídrico e osmocondicionamento de sementes de espécies e híbridos de eucalipto

Pós-graduação em Agronomia (Genética e Melhoramento de Plantas) - FCAV

GERMINATION OF EUCALYPTUS SEEDS UNDER WATER AND SALT STRESS

The knowledge of the physiology of Eucalyptus spp. germination may contribute significantly to the development of management and choice of suitability of the deployment areas. The aim was to evaluate the effects of water and salt stress on seed germination of Eucalyptus camaldulensis, E. citriodora, E. grandis, E. robusta and E. urophylla. The seeding was done with four replicates of 0.05 g of seeds in paper moistened with solutions at potentials of 0.0, -0.2, -0.4, and -0.8 MPa, induced with polyethylene glycol (PEG 6000) and NaCl. The germination test was in 25 degrees C in the presence of light. Were evaluated the first test score seven days after sowing, and weekly germination (normal seedlings) until 28 days. Were also calculated the germination speed index. Water stress causes a greater reduction in the rate of germination and accumulated germination of E. camaldulensis and E. citriodora seeds than salt stress, and the seeds of E. robusta are more adapted to germinate under salt stress moderate, between -0.2 and -0.4 MPa. Regardless of the substance used to induce stress, the threshold for germination was -0.8 MPa. The E. camaldulensis is the most sensitive specie to water stress and E. urophylla most sensitive to salt stress.

Estudos estruturais com a importina-α do fungo Neurospora crassa e sequências de localização nuclear

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

Priming and temperature limits for germination of dispersal units of Urochloa brizantha (Stapf) Webster cv. basilisk

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Effect of mucoadhesive polymers on the in vitro performance of insulin-loaded silica nanoparticles: Interactions with mucin and biomembrane models

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

Determinação das temperaturas cardeais para a germinação de sementes de Murraya paniculata L. Jack. (Rutaceae)

A Murraya paniculata, é conhecida popularmente como murta-de-cheiro ou falsamurta, pertence à família Rutaceae e é uma espécie nativa da Índia que foi introduzida no Brasil. Essa espécie é utilizada na arborização de ruas e em jardins das cidades graças à copa densa e sua resistência a condições adversas. Pode ser utilizada também para formação de cerca vivas e, sua madeira branca tem alta durabilidade podendo ser utilizada em marcenarias. Na Ásia, esta espécie é considerada medicinal e as folhas e raízes são utilizadas para o tratamento de problemas intestinais, de reumatismo e tosse. Do ponto de vista químico, esta espécie acumula principalmente cumarinas e flavonóides, além de derivados do ácido cinâmico e alcalóides. No presente trabalho propomos determinar as temperaturas cardeais para a germinação de sementes de Murraya paniculata, além disso, analisamos a influência do teor de água na germinação para determinar se as sementes são recalcitrantes ou ortodoxas, e a influencia do potencial hídrico induzido por polietilenoglicol (PEG 6000). O aprofundamento dos estudos dessa espécie se faz importante por conta desta ser hospedeira alternativa do psilídeo Diaphorina citri, transmissor do ―greening‖ dos citros, doença que diminuiu a produção de citros nos últimos anos

Osmotic potentials on water uptake and germination of Guazuma ulmifolia Lam. (Sterculiaceae) seeds

Osmotic potentials on water uptake and germination of Guazuma Ulmifolia Lam. (Sterculiaceae) seeds. This work was carried out in the Germination Lab. of the Department of Botany, Institute of Biosciences, São Paulo State University (UNESP), Botucatu, São Paulo State, Brazil. The aims of this work were to determine the water uptake curve and to evaluate the germination of Guazuma ulmifolia seeds subjected to different water potentials. For the water uptake curve, seven replicates of 50 pre-scarified seeds were placed onto paper moistened with 15 mL PEG 6000 solution under the potentials 0 (control), -0.3 and -0.6 MPa at 25o C in the darkness. For the germination assay, four replicates of 50 seeds were subjected to the same above-described conditions; however, one lot of seeds was modified when there was variation in the refractometric index, whereas the remaining ones were kept in the same solutions until the end of the experiment. All three phases of water uptake were detected under 0 and -0.3 MPa; however, phase II was prolonged under -0.6 MPa and germination was not observed. For 0 and -0.3 MPa, the adopted statistical models consisted of asymptotic (phases I and II) and exponential (phase III) functions, y = a*[1 - b*exp (-c*t) + exp (-d + e*(t - t0)]. For -0.6MPa, only the asymptotic function y = a* [1 - b* exp (-c*t)] was used since there was no evidence of germination. The germination final percentage and speed index were lower under -0.3 MPa, mainly when solutions were not replaced; besides, germination was not detected under -0.6 MPa, with or without solution replacement.

DEVELOPMENT AND VALIDATION OF A HPLC METHOD FOR QUANTIFICATION OF URSOLIC ACID IN SOLID DISPERSIONS

DEVELOPMENT AND VALIDATION OF A HPLC METHOD FOR QUANTIFICATION OF URSOLIC ACID IN SOLID DISPERSIONS. Ursolic acid is a natural molecule that presents several pharmacological properties. In this work, an analytical method by RP-HPLC has been developed and validated for quantification of this drug in the solid dispersions, using PEG 6000 and Poloxamer 407 as polymers. The method was specific, linear in the range of 1.0-50.0 mu g mL(-1) (r<0.99), precise (CV < 5% for both inter-and intra-assays), accurate (maximum deviation of +/- 13%), and robust to the parameters evaluated. This method has proved to be simple and useful for ursolic acid determination in solid dispersions, enabling its determination in pharmaceutical dosage form.

Desenvolvimento e validação de um método analítico por CLAE para quantificação de ácido ursólico em dispersões sólidas

Ursolic acid is a natural molecule that presents several pharmacological properties. In this work, an analytical method by RP-HPLC has been developed and validated for quantification of this drug in the solid dispersions, using PEG 6000 and Poloxamer 407 as polymers. The method was specific, linear in the range of 1.0-50.0 µg mL-1 (r<0.99), precise (CV < 5% for both inter- and intra-assays), accurate (maximum deviation of ± 13%), and robust to the parameters evaluated. This method has proved to be simple and useful for ursolic acid determination in solid dispersions, enabling its determination in pharmaceutical dosage form.

Hydric restriction as vigor assessment method of soybean seeds

The objective of this study was to investigate the possibility of using hydric restriction as a method for evaluating vigor of soybean seeds. The soybean seeds, cultivar BRS 245RR, represented by four different seed lots, were characterized by germination and vigor. For the treatment of hydric restriction and temperature, the combination of substrate water potential and temperature were the following: deionized water (0.0 MPa); polyethylene glycol (PEG 6000) aqueous solution (-0.1, -0.3 and -0.5 MPa); and four temperatures (20 ºC, 25 ºC, 30 ºC, and 35 ºC), respectively. A completely randomized experimental design was used, with four replications per treatment, and the ANOVA was performed individually for each combination of temperature and water potential of substrate. According to results obtained, the test of hydric restriction has the same efficiency of the accelerated aging test in estimating vigor of soybean seeds, cv. BRS 245RR, when water potentials of -0.1 MPa or -0.3 MPa at a temperature of 25 ºC, or -0.3 MPa at a temperature of 30 ºC are used.

Implications of macromolecular crowding for signal transduction and metabolite channeling

The effect of different total enzyme concentrations on the flux through the bacterial phosphoenolpyruvate:carbohydrate phosphotransferase system (PTS) in vitro was determined by measuring PTS-mediated carbohydrate phosphorylation at different dilutions of cell-free extract of Escherichia coli. The dependence of the flux on the protein concentration was more than linear but less than quadratic. The combined flux–response coefficient of the four enzymes constituting the glucose PTS decreased slightly from values of ≈1.8 with increasing protein concentrations in the assay. Addition of the macromolecular crowding agents polyethylene glycol (PEG) 6000 and PEG 35000 led to a sharper decrease in the combined flux–response coefficient, in one case to values of ≈1. PEG 6000 stimulated the PTS flux at lower protein concentrations and inhibited the flux at higher protein concentrations, with the transition depending on the PEG 6000 concentration. This suggests that macromolecular crowding decreases the dissociation rate constants of enzyme complexes. High concentrations of the microsolute glycerol did not affect the combined flux–response coefficient. The data could be explained with a kinetic model of macromolecular crowding in a two-enzyme group-transfer pathway. Our results suggest that, because of the crowded environment in the cell, the different PTS enzymes form complexes that live long on the time-scale of their turnover. The implications for the metabolic behavior and control properties of the PTS, and for the effect of macromolecular crowding on nonequilibrium processes, are discussed.

Análise in vitro da expressão de genes de resposta a estresse oxidativo e osmótico da bactéria fastidiosa Leifsonia xyli subsp. xyli

Atualmente, o Brasil é o maior produtor de cana-de-açúcar (Saccharum ssp.), no qual o estado de São Paulo é responsável por mais de 50% da produção. Esta cultura é hospedeira de diversos patógenos que podem limitar sua produção, dentre os quais se destaca a bactéria Leifsonia xyli subsp. xyli (Lxx), agente causal do raquitismo da soqueira (ratoon stunting disease - RSD). Pouco se sabe sobre a fisiologia deste organismo e quais as estratégias utilizadas por este para colonizar seu hospedeiro. No entanto, sabemos que para infectar e colonizar seus hospedeiros, é necessário que bactérias parasíticas superem estresses de diversas naturezas impostas durante estes processos, como os estresses oxidativo e o osmótico. Neste contexto, os objetivos deste trabalho foram identificar in silico e analisar a expressão in vitro, por qPCR, de genes relacionados a estes dois estresses. Uma análise da sequência do genoma de Lxx identificou 35 genes, sendo 8 relacionados ao estresse oxidativo, 9 relacionados ao estresse osmótico e 11 relacionados a estresse gerais, incluindo um cluster de 6 genes envolvidos na síntese de carotenoides. A expressão destes foi avaliada 60 minutos após exposição a 30mM de H2O2 ou 7% (p/v) de polietilenoglicol 6000 (PEG 6000). Sete genes foram avaliados como normalizadores das reações de qPCR. A quantificação do grau de peroxidação lipídica indicou que ambos os tratamentos resultaram em sensível peroxidação, muito embora o efeito do tratamento com PEG 6000 tenha sido maior do que o tratamento com H2O2. A exposição ao H2O2 aumentou a expressão dos genes katA (catalase), sodA (superóxido dismutase), msrA (Sulfóxido de metionina redutase) e msrB (Sulfóxido de metionina redutase) bem como de todos os genes responsáveis pela síntese de carotenoides. Por outro lado, todos os genes relacionados ao estresse osmótico foram menos expressos na presença deste composto. Já quando a bactéria foi exposta a PEG 6000, o oposto ocorreu, ou seja, os genes relacionados ao estresse osmótico, que são otsA (Trealose-6-fosfato sintase), otsB (Trealose fosfatase), treY (Malto-oligosil trealose sintase), treZ (Malto-oligosil trealose trealoidrolase), treS (Trealose sintase), proX (Proteína de ligamento em substrato, tipo ABC glicina betaína transportadora), proW (Proteína permease, tipo ABC glicina betaína transportadora), proZ (Proteína permease, tipo ABC glicina betaína transportadora) e Naggn (Amidotransferase), além dos genes do cluster carotenoide, foram mais expressos, ao passo que alguns dos genes ligados à resposta ao estresse oxidativo foram menos expressos. Verificou-se também, através de PCR convencional utilizando primers para amplificar as regiões entre os genes carotenoides, que estes são expressos como um RNA policistrônico, constituindo assim um operon. Estes resultados validam predições anteriores baseadas na análise in silico da sequência do genoma de Lxx, confirmando que Lxx possui mecanismos responsivos aos estresses osmótico e oxidativo aos quais é submetida durante o processo de infecção de seu hospedeiro.

Alteration of cell-wall porosity is involved in osmotic stress-induced enhancement of aluminium resistance in common bean (Phaseolus vulgaris L.)

Aluminium (Al) toxicity and drought are the two major abiotic stress factors limiting common bean production in the tropics. Using hydroponics, the short-term effects of combined Al toxicity and drought stress on root growth and Al uptake into the root apex were investigated. In the presence of Al stress, PEG 6000 (polyethylene glycol)-induced osmotic (drought) stress led to the amelioration of Al-induced inhibition of root elongation in the Al-sensitive genotype VAX 1. PEG 6000 (>> PEG 1000) treatment greatly decreased Al accumulation in the 1 cm root apices even when the roots were physically separated from the PEG solution using dialysis membrane tubes. Upon removal of PEG from the treatment solution, the root tips recovered from osmotic stress and the Al accumulation capacity was quickly restored. The PEG-induced reduction of Al accumulation was not due to a lower phytotoxic Al concentration in the treatment solution, reduced negativity of the root apoplast, or to enhanced citrate exudation. Also cell-wall (CW) material isolated from PEG-treated roots showed a low Al-binding capacity which, however, was restored after destroying the physical structure of the CW. The comparison of the Al(3+), La(3+), Sr(2+), and Rb(+) binding capacity of the intact root tips and the isolated CW revealed the specificity of the PEG 6000 effect for Al. This could be due to the higher hydrated ionic radius of Al(3+) compared with other cations (Al(3+) >> La(3+) > Sr(2+) > Rb(+)). In conclusion, the results provide circumstantial evidence that the osmotic stress-inhibited Al accumulation in root apices and thus reduced Al-induced inhibition of root elongation in the Al-sensitive genotype VAX 1 is related to the alteration of CW porosity resulting from PEG 6000-induced dehydration of the root apoplast.

GERMINAÇÃO DE SEMENTES DE Piptadenia moniliformis Benth. SOB ESTRESSE HÍDRICO

O período de germinação e o estabelecimento de plântulas é um dos fatores mais importantes para a sobrevivência das espécies, principalmente nos locais em que a disponibilidade de água é limitada, como na região da Caatinga. Neste sentido, o objetivo deste trabalho foi avaliar o efeito do estresse hídrico sobre a germinação de sementes de Piptadenia moniliformis Benth. Foram utilizados três lotes (L1, L2 e L3), correspondentes aos anos de produção de 2006, 2007 e 2008, respectivamente. Antes do teste de germinação, as sementes foram escarificadas com ácido sulfúrico concentrado durante 30 minutos. Para induzir o deficit hídrico, foi utilizado o polietileno glicol (PEG 6000), nos seguintes potenciais osmóticos: - 0,3; -0,6; -0,9, -1,2 e -1,5 MPa e a água (0 MPa) sob as temperaturas de 25 e 30ºC. As características avaliadas foram: porcentagem de germinação e de plântulas normais, índice de velocidade de germinação e massa seca de plântulas. O processo germinativo de sementes de Piptadenia moniliformis Benth. é comprometido a partir de potenciais hídricos inferiores a -0,6 MPa a 25 e 30 °C; potenciais hídricos iguais ou inferiores a -1,2 MPa inibem a formação de plântulas normais nas duas temperaturas; a tolerância ao estresse hídrico simulado com PEG 6000 é variável entre lotes de sementes e temperaturas de germinação.