Proteomic analysis reveals suppression of bark chitinases and proteinase inhibitors in citrus plants affected by the citrus sudden death disease

Citrus sudden death (CSD) is a disease of unknown etiology that greatly affects sweet oranges grafted on Rangpur lime rootstock, the most important rootstock in Brazilian citriculture. We performed a proteomic analysis to generate information related to this plant pathogen interaction. Protein profiles from healthy, CSD-affected and CSD-tolerant stem barks, were generated using two-dimensional gel electrophoresis. The protein spots were well distributed over a pI range of 3.26 to 9.97 and a molecular weight (MW) range from 7.1 to 120 kDa. The patterns of expressed proteins on 2-DE gels made it possible to distinguish healthy barks from CSD-affected barks. Protein spots with MW around 30 kDa and pI values ranging from 4.5 to 5.2 were down-regulated in the CSD-affected rootstock bark. This set of protein spots was identified as chitinases. Another set of proteins, ranging in pI from 6.1 to 9.6 with an MW of about 20 kDa, were also suppressed in CSD-affected rootstock bark; these were identified as miraculin-like proteins, potential trypsin inhibitors. Downregulation of chitinases and proteinase inhibitors in CSD-affected plants is relevant since chitinases are well-known pathogenesis-related protein, and their activity against plant pathogens is largely accepted.

Thermal behavior and structural properites of plant-derived eugenyl acetate

Eugenol is an allyl chain-substituted guaiacol in the biosynthesized phenylpropanoid compound class derived from Syzygium aromaticum L. and widely used in folk medicine. Nonetheless, its pharmacological use is limited by some problems, such as instability when exposed to light and high temperature. In order to enhance stability, the eugenol molecule was structurally modified, resulting in eugenyl acetate. The eugenyl acetate`s thermal behavior and crystal structure was then characterized by differential scanning calorimetry (DSC) and X-ray diffraction (XRD) and compared to a commercial sample.

Energy Analysis within Industrial Hydraulics and Correspondent Solar PV System Design

Energy efficiency and renewable energy use are two main priorities leading to industrial sustainability nowadays according to European Steel Technology Platform (ESTP). Modernization efforts can be done by industries to improve energy consumptions of the production lines. These days, steel making industrial applications are energy and emission intensive. It was estimated that over the past years, energy consumption and corresponding CO2 generation has increased steadily reaching approximately 338.15 parts per million in august 2010 [1]. These kinds of facts and statistics have introduced a lot of room for improvement in energy efficiency for industrial applications through modernization and use of renewable energy sources such as solar Photovoltaic Systems (PV).The purpose of this thesis work is to make a preliminary design and simulation of the solar photovoltaic system which would attempt to cover the energy demand of the initial part of the pickling line hydraulic system at the SSAB steel plant. For this purpose, the energy consumptions of this hydraulic system would be studied and evaluated and a general analysis of the hydraulic and control components performance would be done which would yield a proper set of guidelines contributing towards future energy savings. The results of the energy efficiency analysis showed that the initial part of the pickling line hydraulic system worked with a low efficiency of 3.3%. Results of general analysis showed that hydraulic accumulators of 650 liter size should be used by the initial part pickling line system in combination with a one pump delivery of 100 l/min. Based on this, one PV system can deliver energy to an AC motor-pump set covering 17.6% of total energy and another PV system can supply a DC hydraulic pump substituting 26.7% of the demand. The first system used 290 m2 area of the roof and was sized as 40 kWp, the second used 109 m2 and was sized as 15.2 kWp. It was concluded that the reason for the low efficiency was the oversized design of the system. Incremental modernization efforts could help to improve the hydraulic system energy efficiency and make the design of the solar photovoltaic system realistically possible. Two types of PV systems where analyzed in the thesis work. A method was found calculating the load simulation sequence based on the energy efficiency studies to help in the PV system simulations. Hydraulic accumulators integrated into the pickling line worked as energy storage when being charged by the PV system as well.

Atividade antiplasmódica e toxicológica de plantas medicinais usadas popularmente no Brasil : uma abordagem etnobotânica

Malaria is a disease of global distribution, recognized by governments around the world as a serious public health problem, affecting more than 109 countries and territories and endangering more than 3.3 billion people. The economic costs of this disease are also relevant: the African continent itself has malaria-related costs of about $ 12 billion annually. Nowadays, in addition to chloroquine, Plasmodium falciparum is resistant to many drugs used in the treatment of malaria, such as amodiaquine, mefloquine, quinine and sulfadoxine-pyrimethamine; resistance of Plasmodium vivax to treatments, although less studied, is also reported. Nature, in general, is responsible for the production of most known organic substances, and the plant kingdom is responsible for the most of the chemical diversity known and reported in the literature. Most medicinal plants commercialized in Brazil, however, are of exotic origin, which makes the search for endemic medicinal plants, besides a patent necessity, a fascinating subject of academic research and development. This study aimed to: (i) verify the antimalarial activity of ethanolic and hydroalcoholic extracts of Boerhavia paniculata Rich. And acetonic extract of Clethra scabra Pers. in Swiss albino mice infected by Plasmodium berghei NK65, (ii) observe possible combined effects between the course of infection by P. berghei NK65 and administration of these extracts in Swiss albino mice, and (iii) conduct a preliminary study of the acute toxicity of these extracts in Swiss albino mice. All extracts notable pharmacological activities - with parasite infections inhibitions ranging from 22% to 54%.These characteristics suggest that the activities are relevant, although comparatively lower than the activity displayed by the positive control group (always above 90%). The general framework of survival analysis demonstrates an overall reduction in survival times for all groups. Necroscopy has not pointed no change in color, shape, size and/or consistency in the evaluated organs - the only exception was the livers of rats submitted to treatment to hydroalcoholic extracts: these organs have been presented in a slightly congestive aspect with mass increasing roughly 28% higher than the other two groups and a p-value of 0.0365. The 250 mg/Kg ethanolic group has been pointed out by the Dunn s post test, as the only class with simultaneous inequalities (p<0.05) between positive and negative control groups. The extracts, notably ethanol extract, have, in fact, a vestigial antimalarial activity, although well below from the ones perceived to chloroquine-treated groups; nevertheless, the survival times of the animals fed with the extracts do not rise by presence of such therapy. Both the toxicopharmacological studies of the synergism between the clinical course of malaria and administration of extracts and the isolated evaluation of toxicity allow us to affirm the absence of toxicity of the extracts at the level of CNS and ANS, as well as their non-influence on food and water consumption patterns, until dosages of 500 mg/Kg. Necroscopic analysis leads us to deduct a possible hepatotoxic effect of hydroalcoholic extract at dosages of 500 mg/Kg, and an innocuous tissue activity of the ethanol extract, in the same dosage. We propose a continuation of the studies of these extracts, with protocol modifications capable of addressing more clearly and objectively their pharmacological and toxicological aspects

Functional XPB/RAD25 redundancy in Arabidopsis genome: characterization of AtXPB2 and expression analysis

The xeroderma pigmentosum complementation group B (XPB) protein is involved in both DNA repair and transcription in human cells. It is a component of the transcription factor IIH (TFIIH) and is responsible for DNA helicase activity during nucleotide (nt) excision repair (NER). Its high evolutionary conservation has allowed identification of homologous proteins in different organisms, including plants. In contrast to other organisms, Arabidopsis thaliana harbors a duplication of the XPB orthologue (AtXPB1 and AtXPB2), and the proteins encoded by the duplicated genes are very similar (95% amino acid identity). Complementation assays in yeast rad25 mutant strains suggest the involvement of AtXPB2 in DNA repair, as already shown for AtXPB1, indicating that these proteins may be functionally redundant in the removal of DNA lesions in A. thaliana. Although both genes are expressed in a constitutive manner during the plant life cycle, Northern blot analyses suggest that light modulates the expression level of both XPB copies, and transcript levels increase during early stages of development. Considering the high similarity between AtXPB1 and AtXPB2 and that both of predicted proteins may act in DNA repair, it is possible that this duplication may confer more flexibility and resistance to DNA damaging agents in thale cress. (C) 2004 Elsevier B.V. All rights reserved.

Mutagênese induzida por flavonóides presentes do decocto das cascas da aroeira (Schinus terebinthifolius, Raddi)

The decoction of Brazilian pepper tree barks (Schinus terebinthifolius, Raddi), is used in medicine as wound healing and antiinflamatory. Once extracts from this plant are used for acceleration of scar s process, it is important to study their mutagenic and genotoxic potential. In previous works in our laboratory, it was observed mutagenicity caused by the decoction when in high concentrations. Among the chemical compounds of this plant that could be able to induce mutation, the flavonoids were the only group that was referred to have either an oxidant or antioxidant potential. The flavonoids were isolated, purified and quantified by adsorptive column chromatography under silica gel, bacterial and in vitro genotoxic tests were realized to determine if the flavonoids were the responsible agents for this mutagenicity found. The tests realized with plasmidial DNA were indicative that the flavonoids are probably genotoxic, due to the presence of correlation between increase of the flavonoid concentration and in plasmidial DNA double strand breakage visualized in agarose gel, as well as they were capable to generated abasic sites shown by the in vitro treatment with exonuclease III. The same tests with plasmidial DNA in the presence of copper [10 µM] and of a Tris-HCl pH 7.5 [10 µM] buffer were realized with the isolated flavonoids to determine if there would be or not participation of reactive oxygen species (ROS). The transformation of plasmidial DNA in different bacterial strains proficient and deficient in DNA repair enzymes in the presence or not of a Tris-HCl buffer, suggests that the enzymes that repair oxidative lesions are necessary to repair the lesions generated by the flavonoids and that ROS are generated and are necessary to promote the lesions. Bacterial tests with Escherichia coli strains of the CC collection (deficient or not for DNA repair enzymes), showed that the flavonoids are able to increase the frequency of mutations, mainly in strains mutated in repair enzymes (MutM, MutY-glicosylases and double mutant), suggesting that these agents are responsible for the enhancement in the mutation rate. In order to determine the mutation spectrum caused by the flavonoids of the Brazilian pepper tree stem bark, plasmidial DNA previously treated with the flavonoids were transformed in bacterial strains deficient and proficient in the DNA repair enzymes, followed by a blue-white selection with X-gal, DNA amplification by PCR and sequencing the positive mutant clones. Analysis of the mutants obtained from strains CC104, CC104mutM, CC104mutY, CC104mutMmutY, BW9101, BW9109 indicated a predominance of some mutations like G:C to C:G that can be correlated with the origin of 8-oxoG, due to oxidative lesions caused by the flavonoids. So it can concluded that the flavonoid isolated or in fractions enriched on them are genotoxic and mutagenic, and their mutations are predominantly oxidative, mediated by ROS, and the lesions are recognized by the BER system. In this way it is proposed that the flavonoids can act in two different ways to generate the DNA lesion: 1. in a Fenton-like reaction, when the flavonoid are in the presence of metal ions and that together with the water generate ROS that promotes the DNA lesions; 2. in another way the lesions can be generated by the formation of ROS due to the internal chemical structure of the flavonoid molecule due to the quantity and location of hydroxyl groups, and so producing the DNA lesions, those lesions can be directly (suggested by the in vitro experiments) or indirectly done (supported by the experiments using the CC bacterial strains)

COMBUSTION and PYROLYSIS of A SLUDGE FROM WASTEWATER TREATMENT PLANT

The possibility of thermal treatment plants of municipal wastewater is an alternative solution for the final disposition of the sludge produced on small cities as Barueri, a small town of São Paulo State, Brazil. Combustion and pyrolysis of that municipal waste, occurring respectively in air and nitrogen, have been studied by thermogravimetry (TG) and differential thermal analysis (DTA). The main steps of each case were analyzed and Kissinger plots were used to estimate respective activation energies. DTG peaks are more indicated to represent the condition of maximum reaction rates than DTA peaks.

Plant or fungal sequences? An alternative optimized PCR protocol to avoid ITS (nrDNA) misamplification

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

Xylella fastidiosa: An in vivo system to study possible survival strategies within citrus xylem vessels based on global gene expression analysis

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Effects of plant density and proportion on the interaction between wheat with alexandergrass plants

A determinação das relações competitivas entre espécies de plantas requer delineamentos experimentais e métodos de análise apropriados. A hipótese da pesquisa foi que duas espécies crescendo em convivência têm comportamento de crescimento e desenvolvimento distintos em função da sua competitividade relativa. O objetivo desta pesquisa foi quantificar a competitividade relativa da cultura do trigo com o capim-marmelada através da medida dos efeitos da densidade e da proporção de plantas, usando experimentos em série substitutiva. Monoculturas foram cultivadas em densidades de 1, 3, 5, 10 e 15 plantas por caixa e analisadas por regressão dos dados de massa seca, em 2006. Experimento em mistura foi cultivado nas proporções trigo:capim-marmelada de 0:6, 1:5, 2:4, 3:3, 4:2, 5:1 e 6:0 plantas por caixa e analisado através de interpretação gráfica de características de crescimento e produção, em 2007. Ambos os experimentos foram realizados em delineamento completamente casualizado com quatro repetições. Capim-marmelada foi mais sensível que trigo à competição intraespecífica. Capim-marmelada foi levemente mais competitivo que trigo. Número e massa de espigas e número de afilhos foram as características do trigo mais afetadas pela interferência do capim-marmelada.

Growth analysis of sweet pepper cultivated in coconut fiber in a greenhouse

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

Economic analysis of cucumber and lettuce intercropping under greenhouse in the winter-spring

A análise econômica complementa a avaliação da eficiência dos cultivos consorciados, considerando além da produção física das culturas, o preço dos produtos segundo sua classificação qualitativa e época do ano. Avaliou-se economicamente consórcios de alface crespa e pepino, em duas populações de plantas, no inverno-primavera, em casa de vegetação, em Jaboticabal, SP, Brasil. Foram calculados o custo operacional total (COT), a receita bruta (RB) e o lucro operacional (LO) das culturas do pepino e alface em monocultura e em consórcio. A mão-de-obra foi a componente com maior participação no COT das culturas em consórcio e monocultura. Maiores RB e LO foram observadas nos cultivos consorciados estabelecidos com o transplante da alface e do pepino no mesmo dia, independente da densidade populacional. Considerando-se o lucro operacional e o índice de uso eficiente da área, recomenda-se o cultivo consorciado com transplante da alface até 10 dias após o transplante (DAT) do pepino, com duas linhas, e ao 0 DAT do pepino cultivado com uma linha no canteiro

CABBAGE GROWTH and PRODUCTION IN RELATION TO PLANT DENSITY and NITROGEN

At present, the Brazilian market prefers cabbage (Brassica oleracea var. capitata) of smaller size, which can be achieved by increasing population density; yet this management can alter the optimum rate of nitrogen (N), its second most required nutrient. This study was conducted in the Universidade Estadual Paulista, Jaboticabal, São Paulo, Brazil, from February to June 2004. The objective was to evaluate the effect of population density: 31 250 (DI) and 46 875 plants ha(-1) (D2) with 0, 100, 200 and 300 kg N ha-1 on the growth and production of cabbage Astrus. The experimental design consisted of randomized blocks with a 2 x 4 factorial arrangement and three replicates. Statistical analysis were a variance analysis (F test), the Tukey test for population density averages and polynomial regression for the N rates. In D1, an increase was recorded in the number of inner and outer leaves, dry matter of inner and outer leaves, the stem diameter at the insertion of the head, and the stem dry and fresh matter. The maximum size of the plant in D2 was 1.57 kg and was obtained with 300 kg N ha(-1), while in D1 it was 2.1 kg and was obtained with 244 kg N ha(-1). The optimal economic rate in 131 was 227.1 kg N ha(-1). The highest yield (72.7 t ha(-1)) was obtained with the highest N rate in D2. Smaller cabbage heads, commercially preferred, were obtained without the application of N, regardless of plant population.

Genomics-based design of defined growth media for the plant pathogen Xylella fastidiosa

Based on the genetic analysis of the phytopathogen Xylella fastidiosa genome, five media with defined composition were developed and the growth abilities of this fastidious prokaryote were evaluated in liquid media and on solid plates. All media had a common salt composition and included the same amounts of glucose and vitamins but differed in their amino acid content. XDM1 medium contained amino acids threonine, serine, glycine, alanine, aspartic acid and glutamic acid, for which complete degradation pathways occur in X fastidiosa; XDM2 included serine and methionine, amino acids for which biosynthetic enzymes are absent, plus asparagine and glutamine, which are abundant in the xylem sap; XDM3 had the same composition as XDM2 but with asparagine replaced by aspartic acid due to the presence of complete degradation pathway for aspartic acid; XDM4 was a minimal medium with glutamine as a sole nitrogen source; XDM5 had the same composition as XDM4, plus methionine. The liquid and solidified XDM2 and XDM3 media were the most effective for the growth of X. fastidiosa. This work opens the opportunity for the in silico design of bacterial defined media once their genome is sequenced. (C) 2002 Federation of European Microbiological Societies. Published by Elsevier B.V. B.V. All rights reserved.

Analysis of gene expression profiles under water stress in tolerant and sensitive sugarcane plants

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