Experimental pressure versus temperature isochoric – isoplethic curves for n-pentane – dimethyl ether, n-pentane – dimethyl ether – polybutadiene and n-pentane – dimethyl ether – polybutadiene– hydrogen at high pressures

Loci of isochoric - isoplethic experimental phase equilibrium data, were determined for the binary mixture dimethyl ether (DME) + n-pentane (C5); the ternary mixture: DME + C5 + polybutadiene (PB); and the quaternary mixture DME + C5 + PB + hydrogen (H2). Binary experiments were performed at varying overall density (ρ) and varying quantity of C5. Ternary experiments were performed at varying ρ and varying relative quantities of each light solvent. In the case of quaternary mixtures, the mass fraction of polymer was kept constant, and the amount of H2 and ρ were varied. The experimental data obtained for binary and ternary mixtures were correlated using the Perturbed-Chain Statistical Associating Fluid Theory (PC-SAFT) equation of state (EoS)

Transcriptional profile of maize roots under acid soil growth

Aluminum (Al) toxicity is one of the most important yield-limiting factors of many crops worldwide. The primary symptom of Al toxicity syndrome is the inhibition of root growth leading to poor water and nutrient absorption. Al tolerance has been extensively studied using hydroponic experiments. However, unlike soil conditions, this method does not address all of the components that are necessary for proper root growth and development. In the present study, we grew two maize genotypes with contrasting tolerance to Al in soil containing toxic levels of Al and then compared their transcriptomic responses

A Novel Stress-induced Sugarcane Gene Confers Tolerance To Drought, Salt And Oxidative Stress In Transgenic Tobacco Plants.

Drought is a major abiotic stress that affects crop productivity worldwide. Sugarcane can withstand periods of water scarcity during the final stage of culm maturation, during which sucrose accumulation occurs. Meanwhile, prolonged periods of drought can cause severe plant losses. In a previous study, we evaluated the transcriptome of drought-stressed plants to better understand sugarcane responses to drought. Among the up-regulated genes was Scdr1 (sugarcane drought-responsive 1). The aim of the research reported here was to characterize this gene. Scdr1 encodes a putative protein containing 248 amino acids with a large number of proline (19%) and cysteine (13%) residues. Phylogenetic analysis showed that ScDR1is in a clade with homologs from other monocotyledonous plants, separate from those of dicotyledonous plants. The expression of Scdr1 in different varieties of sugarcane plants has not shown a clear association with drought tolerance. The overexpression of Scdr1 in transgenic tobacco plants increased their tolerance to drought, salinity and oxidative stress, as demonstrated by increased photosynthesis, water content, biomass, germination rate, chlorophyll content and reduced accumulation of ROS. Physiological parameters, such as transpiration rate (E), net photosynthesis (A), stomatal conductance (gs) and internal leaf CO(2) concentration, were less affected by abiotic stresses in transgenic Scdr1 plants compared with wild-type plants. Overall, our results indicated that Scdr1 conferred tolerance to multiple abiotic stresses, highlighting the potential of this gene for biotechnological applications.7e4469

Oligomerization, Membrane Association, and in Vivo Phosphorylation of Sugarcane UDP-glucose Pyrophosphorylase

Sugarcane is a monocot plant that accumulates sucrose to levels of up to 50% of dry weight in the stalk. The mechanisms that are involved in sucrose accumulation in sugarcane are not well understood, and little is known with regard to factors that control the extent of sucrose storage in the stalks. UDP-glucose pyrophosphorylase (UGPase; EC 2.7.7.9) is an enzyme that produces UDP-glucose, a key precursor for sucrose metabolism and cell wall biosynthesis. The objective of this work was to gain insights into the ScUGPase-1 expression pattern and regulatory mechanisms that control protein activity. ScUGPase-1 expression was negatively correlated with the sucrose content in the internodes during development, and only slight differences in the expression patterns were observed between two cultivars that differ in sucrose content. The intracellular localization of ScUGPase-1 indicated partial membrane association of this soluble protein in both the leaves and internodes. Using a phospho-specific antibody, we observed that ScUGPase-1 was phosphorylated in vivo at the Ser-419 site in the soluble and membrane fractions from the leaves but not from the internodes. The purified recombinant enzyme was kinetically characterized in the direction of UDP-glucose formation, and the enzyme activity was affected by redox modification. Preincubation with H2O2 strongly inhibited this activity, which could be reversed by DTT. Small angle x-ray scattering analysis indicated that the dimer interface is located at the C terminus and provided the first structural model of the dimer of sugarcane UGPase in solution.Fil: Soares, Jose Sergio M.. Universidade Estadual de Campinas. Instituto de Biologia. Departamento de Genética, Evolução e Bioagentes; BrasilFil: Gentile, Agustina. Universidade Estadual de Campinas. Instituto de Biologia. Departamento de Genética, Evolução e Bioagentes; BrasilFil: Scorsato, Valeria. Universidade Estadual de Campinas. Instituto de Química. Laboratório de Biologia Estrutural e Cristalografia; BrasilFil: Lima, Aline da C.. Universidade Estadual de Campinas. Instituto de Química. Laboratório de Biologia Estrutural e Cristalografia; BrasilFil: Kiyota, Eduardo. Universidade Estadual de Campinas. Instituto de Química. Laboratório de Biologia Estrutural e Cristalografia; BrasilFil: Santos, Marcelo Leite Dos . Universidade Federal do Sergipe. Centro de Ciências Exatas e Tecnologia. Núcleo de Química; BrasilFil: Piattoni, Claudia Vanesa. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Santa Fe. Instituto de Agrobiotecnologia del Litoral; Argentina. Universidad Nacional del Litoral; ArgentinaFil: Huber, Steven C.. University of Illinois at Urbana-Champaign. Department of Agriculture Agricultural Research Service, and Department of Plant Biology; Estados UnidosFil: Aparicio, Ricardo. Universidade Estadual de Campinas. Instituto de Química. Laboratório de Biologia Estrutural e Cristalografia; BrasilFil: Menossi, Marcelo. Universidade Estadual de Campinas. Instituto de Biologia. Departamento de Genética, Evolução e Bioagentes; Brasi

In silico evaluation of the Eucalyptus transcriptome

The expressed sequence tags (ESTs) produced in the Forests project provide an invaluable opportunity to assess the Eucalyptus transcriptome. Besides providing information on the different proteins produced by this plant, it is possible to infer gene expression profiles because non-normalized cDNA libraries were used. The EST frequency from any gene is correlated to the transcript levels in the tissues from which the cDNA libraries were constructed. The goal of this work was to identify Eucalyptus genes that showed either differential expression pattern or were ubiquitously expressed in the tissues sampled in the Forests project. Six robust statistical tests and very restrictive rules were applied to gain confidence in the in silico data aiming to avoid false positives. Several genes with interesting expression profiles were identified and some of them were validated by RT-PCR.487495Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq

Avaliação da performance de diferentes plásticos usados para selar arranjos de cDNA em náilon

cDNA arrays are a powerful tool for discovering gene expression patterns. Nylon arrays have the advantage that they can be re-used several times. A key issue in high throughput gene expression analysis is sensitivity. In the case of nylon arrays, signal detection can be affected by the plastic bags used to keep membranes humid. In this study, we evaluated the effect of five types of plastics on the radioactive transmittance, number of genes with a signal above the background, and data variability. A polyethylene plastic bag 69 μm thick had a strong shielding effect that blocked 68.7% of the radioactive signal. The shielding effect on transmittance decreased the number of detected genes and increased the data variability. Other plastics which were thinner gave better results. Although plastics made from polyvinylidene chloride, polyvinyl chloride (both 13 μm thick) and polyethylene (29 and 7 μm thick) showed different levels of transmittance, they all gave similarly good performances. Polyvinylidene chloride and polyethylene 29 mm thick were the plastics of choice because of their easy handling. For other types of plastics, it is advisable to run a simple check on their performance in order to obtain the maximum information from nylon cDNA arrays.Os arranjos de cDNA são uma poderosa ferramenta para o estudo de padrões de expressão gênica. Os arranjos em membranas de náilon apresentam ainda a vantagem de poderem ser reutilizados diversas vezes. Porém, um ponto bastante delicado em estudos de expressão gênica em larga escala é a sensibilidade. No caso de arranjos em membranas de náilon, a detecção dos sinais pode ser afetada pelo envoltório plástico utilizado para manter as membranas úmidas. Nesse estudo, nós avaliamos os efeitos de cinco tipos de plásticos na transmissão radioativa detectada, no número de genes com sinal acima da emissão de fundo e na variabilidade dos dados. O plástico produzido com polietileno com 69 μm de espessura apresentou uma forte interferência na emissão radioativa, bloqueando 68.7% do sinal detectado. Este bloqueio na transmitância diminuiu o numero de genes detectados e aumentou a variabilidade dos dados. Outros plásticos mais finos tiveram resultados melhores. Apesar de plásticos feitos de cloreto de polivinilideno e cloreto de polivinila (ambos com 13 μm de espessura) e polietileno (29 e 7 μm de espessura) terem diferentes níveis de transmitância, todos apresentaram performances semelhantes nos testes realizados. Cloreto de polivinilideno e polietileno com 29 μm de espessura foram os plásticos escolhidos devido à facilidade de manuseio. Para outros tipos de plásticos, é recomendável realizar um teste de suas performances antes de utilizá-los para envolver membranas de náilon, de forma a obter o máximo de informação dos experimentos com arranjos de cDNA.18831887Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES

A Novel Stress-Induced Sugarcane Gene Confers Tolerance to Drought, Salt and Oxidative Stress in Transgenic Tobacco Plants

Background: Drought is a major abiotic stress that affects crop productivity worldwide. Sugarcane can withstand periods of water scarcity during the final stage of culm maturation, during which sucrose accumulation occurs. Meanwhile, prolonged periods of drought can cause severe plant losses. Methodology/Principal Findings: In a previous study, we evaluated the transcriptome of drought-stressed plants to better understand sugarcane responses to drought. Among the up-regulated genes was Scdr1 (sugarcane drought-responsive 1). The aim of the research reported here was to characterize this gene. Scdr1 encodes a putative protein containing 248 amino acids with a large number of proline (19%) and cysteine (13%) residues. Phylogenetic analysis showed that ScDR1is in a clade with homologs from other monocotyledonous plants, separate from those of dicotyledonous plants. The expression of Scdr1 in different varieties of sugarcane plants has not shown a clear association with drought tolerance. Conclusions/Significance: The overexpression of Scdr1 in transgenic tobacco plants increased their tolerance to drought, salinity and oxidative stress, as demonstrated by increased photosynthesis, water content, biomass, germination rate, chlorophyll content and reduced accumulation of ROS. Physiological parameters, such as transpiration rate (E), net photosynthesis (A), stomatal conductance (gs) and internal leaf CO2 concentration, were less affected by abiotic stresses in transgenic Scdr1 plants compared with wild-type plants. Overall, our results indicated that Scdr1 conferred tolerance to multiple abiotic stresses, highlighting the potential of this gene for biotechnological applications.National Council for Scientific and Technological Development (CNPq)National Council for Scientific and Technological Development (CNPq)Fundacao de Amparo a Pesquisa de Sao Paulo (FAPESP)Fundacao de Amparo a Pesquisa de Sao Paulo (FAPESP) [2008/5798-6, 2008/57908-6]Conselho Nacional de Desenvolvimento Cientifico e Tecnologico (CNPq) [574002/2008-1, 552802/2007-7]Conselho Nacional de Desenvolvimento Cientifico e Tecnologico (CNPq)Financiadora de Estudos e Projetos (FINEP) [815/07]Financiadora de Estudos e Projetos (FINEP

Physiological limitations in two sugarcane varieties under water suppression and after recovering

Increasing water scarcity and depleted water productivity in irrigated soils are inducing farmers to adopt improved varieties, such as those with high-capacity tolerance. The use of tolerant varieties of sugarcane might substantially avoid the decline of productivity under water deficit. This research aimed to evaluate the harmful effects of drought on the physiology of two sugarcane varieties (RB867515 and RB962962) during the initial development. Young plants were subjected to irrigation suspension until total stomata closure, and then rewatered. Significant reduction on stomatal conductance, transpiration, and net photosynthesis were observed. RB867515 showed a faster stomatal closure while RB962962 slowed the effects of drought on the gas exchanges parameters with a faster recovering after rewatering. Accumulation of carbohydrates, amino acids, proline, and protein in the leaves and roots of the stressed plants occurred in both varieties, substantially linked to reduction of the leaf water potential. Due to the severity of stress, this accumulation was not enough to maintain the cell turgor pressure, so relative water content was diminished. Water stress affected the contents of chlorophyll (a, b, and total) in both varieties, but not the levels of carotenoids. There was a significant reduction in dry matter under stress. In conclusion, RB962962 variety endured stressed conditions more than RB867515, since it slowed down the damaging effects of drought on the gas exchanges. In addition, RB962962 presented a faster recovery than RB867515, a feature that qualifies it as a variety capable of enduring short periods of drought without major losses in the initial stage of its development.213222Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq

Identification of new ABA-and MEJA-activated sugarcane bZIP genes by data mining in the SUCEST database

Abstract Sugarcane is generally propagated by cuttings of the stalk containing one or more lateral buds, which will develop into a new plant. The transition from the dormant into the active stage constitutes a complex phenomenon characterized by changes in accumulation of phytohormones and several other physiological aspects. Abscisic acid (ABA) and methyl-jasmonate (MeJA) are major signaling molecules, which influence plant development and stress responses. These plant regulators modulate gene expression with the participation of many transcriptional factors. Basic leucine zipper proteins (bZIPs) form a large family of transcriptional factors involved in a variety of plant physiological processes, such as development and responses to stress. Query sequences consisting of fulllength protein sequence of each of the Arabidopsis bZIP families were utilized to screen the sugarcane EST database (SUCEST) and 86 sugarcane assembled sequences (SAS) coding for bZIPs were identified. cDNA arrays and RNA-gel blots were used to study the expression of these sugarcane bZIP genes during early plantlet development and in response to ABA and MeJA. Six bZIP genes were found to be differentially expressed during development. ABA and MeJA modulated the expression of eight sugarcane bZIP genes. Our findings provide novel insights into the expression of this large protein family of transcriptional factors in sugarcane