Método de otimização do processo de produção da goma xantana

O qual se refere a um processo biológico de produção da goma xantana através da propagação celular das bactérias Xanthomonas axonopodis pv. manihotis e/ou Xanthomonas campestris pv campestris e/ou Xanthomonas spp, em cultivos biotecnológicos em condições específicas, bem como etapas de separação e purificação do molécula, sendo que, nessa invenção é utilizado para comprovar a otimização do processo de produção da xantana, parâmetros específicos quantitativo, tais como o rendimento do produto (goma xantana produzida pelo açúcar consumido), aqui simbolizado Yp/s.

Suscetibilidade à podridão negra e produtividade de brócolis em função de doses de nitrogênio e potássio

The purpose of this study was to investigate the influence of potassium and nitrogen on the severity of black rot and yield of head. The experiment was carried out in split plots with factorial subplots, in randomized blocks with four replications. In the plots we assessed the influence of inoculation (injured plants with distilled water and inoculated with Xanthomonas campestris pv. campestris) and in the subplots we evaluated the 4 x 4 factorial, potassium doses (0, 275, 550 and 825 kg ha-1) and nitrogen (0, 150, 300 and 450 kg ha-1). The hybrid Broccoli BRO68 was inoculated using the toothpick method at 64 days after sowing. We observed that potassium in the plant reduced the disease, but nitrogen increased disease. We estimated that the highest levels of potassum in the plant reduced the severity of the disease but lack or excess of nitrogen increased disease severity. The highest production of broccoli inflorescence was obtained at doses from 541 to 751 kg ha-1 of K2O and 286 to 328 kg ha-1 of N. However, when considering a balanced fertilization to obtain higher yield and size of inflorescences and greater resistance to black rot, the ideal doses are 283 kg ha-1 of N and 550 kg ha-1 of K2O.

The copper resistance operon copAB from Xanthomonas axonopodis pathovar citri: gene inactivation results in copper sensitivity

Xanthomonas axonopodis pv. citri (Xac) causes citrus canker and the completion of the Xac genome sequence has opened up the possibility of investigating basic cellular mechanisms at the genomic level. Copper compounds have been extensively used in agriculture to control plant diseases. The copA and copB genes, identified by annotation of the Xac genome, encode homologues of proteins involved in copper resistance. A gene expression assay by Northern blotting revealed that copA and copB are expressed as a unique transcript specifically induced by copper. Synthesis of the gene products was also induced by copper, reaching a maximum level at 4 h after addition of copper to the culture medium. CopA was a cytosolic protein and CopB was detected in the cytoplasmic membrane. The gene encoding CopA was disrupted by the insertion of a transposon, leading to mutant strains that were unable to grow in culture medium containing copper, even at the lowest CUSO4 concentration tested (0.25 mM), whereas the wild-type strain was able to grow in the presence of 1 mM copper. Cell suspensions of the wild-type and mutant strains in different copper concentrations were inoculated in lemon leaves to analyse their ability to induce citrus canker symptoms. Cells of mutant strains showed higher sensitivity than the wild-type strain in the presence of copper, i.e. they were not able to induce citrus canker symptoms at high copper concentrations and exhibited a more retarded growth in planta.

Ativação das vias relacionadas a resistência de Citrus sinensis em resposta a interação com a bactéria Xanthomonas axonopodis

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Síntese, caracterização e avaliação biológica de quelatos metálicos contendo ligantes de origem natural

O trabalho que será aqui apresentado teve como finalidade reunir os resultados obtidos dentro do Laboratório de Organometálicos II em relação aos estudos realizados acerca da interação de íons metálicos com ligantes orgânicos naturais. Os dados apresentados foram obtidos pela autora deste trabalho, e também pelos professores Francisco Otávio Cintra Ferrarini, José Roberto Stefanelli Junior e pela aluna de Iniciação Científica Angélica Romão Carvalho. Hidrolisado proteico de soro de leite bovino e torta de linhaça foram os materiais obtidos, após uma série de operações que envolveram filtração com caulim, diálise, hidrólise enzimática, liofilização, etc., após as quais foram utilizados neste trabalho como ligantes de origem natural. Os metais de transição cobre, ferro, cobalto, zinco, paládio, platina e prata foram utilizados nas reações de complexação com os hidrolisados para obtenção de complexos metálicos, através do uso de diferentes rotas sintéticas (convencional, micro-ondas, hidrotérmica, difusão) e parâmetros experimentais (pH, solvente, natureza do precursor, etc.). A escolha dos íons metálicos baseou-se no interesse em preparar compostos que apresentassem ao mesmo tempo alta biodisponibilidade, baixa citotoxicidade e diversas aplicações biológicas. O uso do cobalto deveu-se à reconhecida propriedade que os micro-organismos do sistema digestório dos ruminantes apresenta de sintetizar a vitamina B12 e, portanto, os compostos desse metal podem ser usados em rações visando um melhor desenvolvimento do animal. O ferro, conhecidamente, é um importante metal para os seres humanos, estando presente na hemoglobina, sendo assim, também consiste em uma substância nutracêutica de relevado interesse. À propósito, cabe salientar que os compostos de ferro (III) obtidos nesse trabalho serão objeto de avaliação do uso como suplemento alimentar, em colaboração com o Departamento de...

Renal and antibacterial effects induced by myotoxin I and II isolated from Bothrops jararacussu venom

Bothrops jararacussu myotoxin I (BthTx-I; Lys 49) and II (BthTX-II; Asp 49) were purified by ion-exchange chromatography and reverse phase HPLC. In this work we used the isolated perfused rat kidney method to evaluate the renal effects of B. jararacussu myotoxins I (Lys49 PLA(2)) and II (Asp49 PLA(2)) and their possible blockage by indomethacin. BthTX-1 (5 mu g/ml) and BthTX-II (5 mu g/ml) increased perfusion pressure (PP; ct(120) = 110.28+/-3.70 mmHg; BthTX I = 171.28+/-6.30* mmHg; BthTX II = 175.50+/-7.20* mmHg), renal vascular resistance (RVR; ct(120) = 5.49+/-0.54 mmHg/ml.g(-1) min(-1); BthTX I = 8.62+/-0.37* mmHg/ml g(-1) min(-1); BthTX II=8.9+/-0.36* mmHg/ml g(-1) min(-1)), urinary flow (UF; ct(120)= 0.14+/-0.01 ml g(-1) min(-1); BthTX I=0.32+/-0.05* ml g(-1) min(-1); BthTX II=0.37+/-0.01* ml g(-1) min(-1)) and glomerular filtration rate (GFR; ct(120)=0.72+/-0.10 ml g(-1) min(-1); BthTX I=0.85+/-0.13* ml g(-1) min(-1); BthTX II=1.22+/-0.28* ml g(-1) min(-1)). In contrast decreased the percent of sodium tubular transport (%TNa+; ct(120)=79,76+/-0.56; BthTX I=62.23+/-4.12*; BthTX II=70.96+/-2.93*) and percent of potassium tubular transport (%TK+;ct(120)=66.80+/-3.69; BthTX I=55.76+/-5.57*; BthTX II=50.86+/-6.16*). Indomethacin antagonized the vascular, glomerular and tubular effects promoted by BthTX I and it's partially blocked the effects of BthTX II. In this work also evaluated the antibacterial effects of BthTx-I and BthTx-II against Xanthomonas axonopodis. pv. passiflorae (Gram-negative bacteria) and we observed that both PLA2 showed antibacterial activity. Also we observed that proteins Also we observed that proteins chemically modified with 4-bromophenacyl bromide (rho-BPB) decrease significantly the antibacterial effect of both PLA(2). In conclusion, BthTx I and BthTX II caused renal alteration and presented activity antimicrobial. The indomethacin was able to antagonize totally the renal effects induced by BthTx I and partially the effects promoted by BthTx II, suggesting involvement of inflammatory mediators in the renal effects caused by myotoxins. In the other hand, other effects could be independently of the enzymatic activity of the BthTX II and the C-terminal domain could be involved in both effects promoted for PLA(2). (C) 2005 Elsevier Ltd. All rights reserved.

Purification and biological effects of a C-type lectin isolated from Bothrops moojeni

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

Mapas de zonas de risco de epidemias e zoneamento agroclimático para o Cancro Cítrico no Estado de São Paulo

O cancro cítrico, causado pela bactéria Xanthomonas axonopodis pv. citri Valterin et alii 1995, é uma doença conhecida mundialmente e sempre constituiu séria ameaça para a citricultura brasileira. O objetivo do presente trabalho foi analisar as condições climáticas do Estado de São Paulo e desenvolver mapas de zonas de maior risco de epidemias de cancro cítrico. Foram utilizados dados meteorológicos referentes aos anos de 2002 a 2005, os quais foram baseados no modelo de previsão desenvolvido por Campbell & Madden (4) e Hau & Kranz (10). A freqüência dos dados foi horária e quando alguma estação apresentava falha, esses eram extrapolados da estação mais próxima. Foram contabilizados os índices de favorabilidade e posteriormente calculadas as porcentagens de dias favoráveis à ocorrência da doença no período de um ano. A partir destas informações, foram gerados os mapas temáticos do Estado de São Paulo, com a distribuição espacial da porcentagem de dias favoráveis à ocorrência de cancro cítrico. A região Noroeste do Estado foi a que apresentou a maior porcentagem de dias favoráveis à ocorrência de cancro cítrico.

Ação de tinturas e óleos essenciais de plantas medicinais sobre o crestamento bacteriano comum do feijoeiro e na produção de proteínas de indução de resistência

A exploração da atividade biológica de compostos secundários presentes nas tinturas ou em óleos essenciais de plantas podem representar, ao lado da indução de resistência, mais uma forma potencial de controle de doenças em plantas cultivadas. O presente trabalho objetivou avaliar o potencial de tinturas de Lippia alba, Lippia sidoides, Mikania glomerata, Equisetum sp. e Hedera helix e óleos essenciais de Rosmarinus officinalis e Cinnamomum zeylanicum nas atividades in vitro, in vivo e na produção de proteínas na indução de resistência, em plantas de feijão vagem cultivar Bragança. Os resultados obtidos demonstraram que as tinturas de L. alba e L. sidoides e os óleos essenciais (R. officinalis e C. zeylanicum) apresentaram atividade in vitro aos isolados de Xanthomonas axonopodis pv. phaseoli. Todas as tinturas ensaiadas apresentaram menores valores do progresso da doença (AACPD), em relação à testemunha, merecendo destaque a tintura de L. alba, que estavam correlacionadas com os maiores teores de polifenoloxidase, peroxidase e proteínas solúveis totais, evidenciando uma possível indução de resistência. Os óleos essenciais não apresentaram diferença na AACPD e nem na indução de proteínas.

Strain variability in the DNA immigration control region (ICR) of Xylella fastidiosa

The genome of the bacterium Xylella fastidiosa contains four ORFs (XF2721, XF2725, XF2739 and XF0295) related to the restriction modification type I system, ordinarily named R-M. This system belongs to the DNA immigration control region (ICR). Each CIRF is related to different operon structures, which are homologues among themselves and with subunit Hsd R from the endonuclease coding genes. In addition, these ORFs are highly homologous to genes in Pseudomonas aeruginosa, Methylococcus capsulatus str. Bath, Legionella pneumophila, Helicobacter pylori, Xanthomonas oryzae pv. Oryzae and Silicibacter pomeroyi, as well as to genes from X. fastidiosa strains that infect grapevine, almond and oleander plants. This study was carried out on R-M ORFs from forty-three X. fastidiosa strains isolated from citrus, coffee, grapevine, periwinkle, almond and plum trees, in order to assess the genetic diversity of these loci through PCR-RFLP. PCR-RFLP analysis of the four ORFs related to the R-M system from these strains enabled the detection of haplotypes for these loci. When the haplotypes were defined, wide genetic diversity and a large range of similar strains originating from different hosts were observed. This analysis also provided information indicating differences in population genetic structures, which led to detection of different levels of gene transfer among the groups of strains. (c) 2005 Elsevier SAS. All rights reserved.

Comparative genomics analyses of citrus-associated bacteria

Xylella fastidiosa 9a5c (XF-9a5c) and Xanthomonas axonopodis pv. citri (XAC) are bacteria that infect citrus plants. Sequencing of the genomes of these strains is complete and comparative analyses are now under way with the genomes of other bacteria of the same genera. In this review, we present an overview of this comparative genomic work. We also present a detailed genomic comparison between XF-9a5a and XAC. Based on this analysis, genes and operons were identified that might be relevant for adaptation to citrus. XAC has two copies of a type II secretion system, a large number of cell wall-degrading enzymes and sugar transporters, a complete energy metabolism, a whole set of avirulence genes associated with a type III secretion system, and a complete flagellar and chemotatic system. By contrast, XF-9a5c possesses more genes involved with type IV pili biosynthesis than does XAC, contains genes encoding for production of colicins, and has 4 copies of Type I restriction/modification system while XAC has only one.

Evaluation of pyraclostrobin and acibenzolar-S-methyl on common bacterial blight of snap bean

Assays were done under greenhouse conditions in order to evaluate the effect of pyraclostrobin (0.0375, 0.0750 and 0.150 mL.L-1) and acibenzolar-S-methyl (ASM) (0.025 g.L-1) in common bacterial blight on leaves of snap beans cultivar Braganca. These chemicals were sprayed at three different times: five days before; five days before + five days after; and five days after leaf inoculation with an isolate of Xanthomonas axonopodis pv. phaseoli. They were determinate the levels of polyphenoloxidase, peroxidase and total soluble proteins on inoculated and non-inoculated leaves of snap beans sprayed with pyraclostrobin (0.075 g.L-1) and ASM (0.025 g.L-1). All concentration of pyraclostrobin and ASM reduced the area under the disease progress curve (AUDPC) on leaves of snap beans, and the least AUDPC value was observed when this products were sprayed five days before + five days after inoculation. Higher levels of polyphenoloxidase, peroxidase and the total soluble proteins were observed on leaves sprayed with pyraclostrobin or ASM.

Combining ability of Phaseolus vulgaris L. for resistance to common bacterial blight

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

Isolation of a new L-amino acid oxidase from Crotalus durissus cascavella venom

A novel L-amino acid oxidase (LAO) (Casca LAO) from Crotalus durissus cascavella venom was purified to a high degree of molecular homogeneity using a combination of molecular exclusion and ion-exchange chromatography system. The purified monomer of LAO presented a molecular mass of 68 kDa and pI estimated in 5.43, which were determined by two-dimensional electrophoresis. The 71st N-terminal amino acid sequence of the LAO from Crotalus durissus cascavella presented a high amino acid sequence similarities with other LAOs from Colloselasma rhosostoma, Crotalus adamanteus, Agkistrodon h. blomhoffi, Agkistrodon h. halys and Trimeresurus stejnegeri. LAO displayed a Michaelis-Menten behavior with a kilometer of 46.7 mu M and an optimum pH for enzymatic activity of 6.5. Casca LAO induced a dose-dependent platelet aggregation, which was abolished by catalase and inhibited by indomethacin and aspirin. These results suggest that the production of H2O2 is involved in subsequent activation of inflammatory enzymes, such as thromboxane. Casca LAO also inhibited the bacterial Growth of Gram-negative (Xanthomonas axonopodis pv passiflorae) and Gram-positive (S. mutans) strains. Electron microscopy assessments of both bacterial strains suggest that the hydrogen peroxide produced by LAO induce bacterial membrane rupture and consequently loss of cytoplasmatic content. This LAO exhibited a high antileishmanic activity against the promastigote of Leishmania amazonensis in vitro, its activity was dependent on the production of hydrogen peroxide, and the 50% inhibitory concentration was estimated in 2.39 mu g/ml. (C) 2005 Elsevier Ltd. All rights reserved.

Effect of temperature and leaf wetness duration on infection of sweet oranges by Asiatic citrus canker

Asiatic citrus canker, caused by Xanthomonas smithii ssp. citri, formerly X. axonopodis pv. citri, is one of the most serious phytosanitary problems in Brazilian citrus crops. Experiments were conducted under controlled conditions to assess the influence of temperature and leaf wetness duration on infection and subsequent symptom development of citrus canker in sweet orange cvs Hamlin, Natal, Pera and Valencia. The quantified variables were incubation period, disease incidence, disease severity, mean lesion density and mean lesion size at temperatures of 12, 15, 20, 25, 30, 35, 40 and 42 degrees C, and leaf wetness durations of 0, 4, 8, 12, 16, 20 and 24 h. Symptoms did not develop at 42 degrees C. A generalized beta function showed a good fit to the temperature data, severity being highest in the range 30-35 degrees C. The relationship between citrus canker severity and leaf wetness duration was explained by a monomolecular model, with the greatest severity occurring at 24 h of leaf wetness, with 4 h of wetness being the minimum duration sufficient to cause 100% incidence at optimal temperatures of 25-35 degrees C. Mean lesion density behaved similarly to disease severity in relation to temperature variation and leaf wetness duration. A combined monomolecular-beta generalized model fitted disease severity, mean lesion density or lesion size as a function of both temperature and duration of leaf wetness. The estimated minimum and maximum temperatures for the occurrence of disease were 12 degrees C and 40 degrees C, respectively.