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.

Reação de híbridos de brócolis 'tipo cabeça única' à podridão negra

Conduziu-se simultaneamente dois experimentos, em casa de vegetação, com o objetivo de avaliar a resistência de híbridos de brócolis 'tipo cabeça única' (AF 649, Titleist, Centenário, Green Power, BR068, Magestic Crown, Marathon, Laguna, Legacy, Green Parasol, Packman e Mônaco) à podridão negra, causada por Xanthomonas campestris pv. campestris. Foram utilizados os métodos de inoculação no ápice das folhas por cortes com tesoura embebida na suspensão bacteriana e por ferimento provocado no caule com palito de dente umedecido na suspensão bacteriana. A inoculação foi realizada aos 25 dias após transplante (6 a 8 folhas definitivas). Avaliou-se no experimento de inoculação com tesoura, as áreas abaixo da curva de progresso da doença nas folhas inoculadas. No experimento de inoculação por palito de dente, avaliou-se a proporção de altura necrosada do caule, aos 26 dias após inoculação. Verificou-se que, em ambos os experimentos, o híbrido BRO68 apresentou-se mais suscetível à podridão negra e os híbridos Marathon, Legacy e Green Power foram os que apresentaram os maiores níveis de resistência à podridão negra.

Grapevine bacterial canker in the State of São Paulo, Brazil: detection and eradication

Sintomas do cancro bacteriano da videira na variedade Red Globe foram observados em agosto de 2009 em pomar de Tupi Paulista, Estado de São Paulo, Brasil, e o agente causal Xanthomonas campestris pv. viticola foi identificado por meio de testes patológicos e moleculares. O procedimento de erradicação foi adotado e aproximadamente 4.700 plantas foram destruídas. Um levantamento realizado nas regiões produtoras do Estado de São Paulo não encontrou nenhum outro pomar contaminado, e essa espécie bacteriana é considerada ausente neste estado.

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.

Desempenho de cultivares de couve-flor de verão em Ipameri

Com o propósito de avaliar o comportamento de seis cultivares de couve-flor de verão, quanto à produção e resistência à podridão negra (Xanthomonas campestris pv. campestris), instalou-se um experimento em campo, em Ipameri-GO, com o delineamento experimental em blocos completos casualizados com seis tratamentos e quatro repetições. Cada parcela experimental, de 10m², constou de 20 plantas distribuídas no espaçamento de 1,0m entre linhas e 0,5m entre plantas. Foram avaliadas as cultivares: híbridos 'Cindy', 'Sarah AF-1169', 'Sharon', 'Verona', 'Lisvera' e a variedade Piracicaba precoce. Os híbridos Verona e Sharon mostraram-se promissores para cultivo nas condições edafoclimáticas em que foram avaliados, com produtividades de 34,17 e 30,64t ha-1, respectivamente, e com moderados níveis de resistência à podridão negra. A variedade Piracicaba precoce apresentou produção inferior às dos melhores híbridos avaliados e moderada resistência à podridão negra. O híbrido Sarah não é recomendado para plantio na região de Ipameri-GO por apresentar baixa produção e baixa resistência à podridão negra.

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

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

Comparative genomic analysis of plant-associated bacteria

This review deals with a comparative analysis of seven genome sequences from plant-associated bacteria. These are the genomes of Agrobacterium tumefaciens, Mesorhizobium loti, Sinorhizobium meliloti, Xanthomonas campestris pv campestris, Xanthomonas axonopodis pv citri, Xylella fastidiosa, and Ralstonia solanacearum. Genome structure and the metabolism pathways available highlight the compromise between the genome size and lifestyle. Despite the recognized importance of the type III secretion system in controlling host compatibility, its presence is not universal in all necrogenic pathogens. Hemolysins, hemagglutinins, and some adhesins, previously reported only for mammalian pathogens, are present in most organisms discussed. Different numbers and combinations of cell wall degrading enzymes and genes to overcome the oxidative burst generally induced by the plant host are characterized in these genomes. A total of 19 genes not involved in housekeeping functions were found common to all these bacteria.

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.

Effects of a lectin-like protein isolated from Acacia farnesiana seeds on phytopathogenic bacterial strains and root-knot nematode

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Virulência de isolados Dexanthomonas campestris pv. Phaseoli (Smith) dye em feijoeiro

The objective of this research was to evaluate de reaction of leaves and pods of five cultivars of bean (Phaseolus vulgaris L.) plants to twenty strains of Xanthomonas campestris pv. phaseoli. The strains were inoculated onto leaves in a greenhouse, and onto pods in a growth chamber. The results obtained were analyzed and the strains classified into three groups: low, medium, and high virulence. Most of the strains showed high virulence on leaves of Carioca and Rio Negro cultivars, as opposed to only low to medium virulence on leaves of IAPAR 14, IAPAR 16, and G. N. Nebraska # 1 sel. 27 cultivars. There were, however, individual strains powerful enough to overcome the leaf resistance of IAPAR 14, IAPAR 16, and G. N. Nebraska # 1 sel. 27. With regard to pods, most strains showed high virulence on all bean cultivars, with exception of IAPAR 14 where virulence was at medium level. A correlation between leaf and pod symptoms was found to exist in Carioca, Rio Negro, and IAPAR 14 cultivars. No such correlation was observed in IAPAR 16 and G. N. Nebraska # 1 sel. 27. Comparing strains producing melanine in vitro with those not producing this pigment, no difference was observed with regard to virulence.

Evaluation of antimicrobial activity of extracts of medicinal plants on three tomato phytopathogens

Antimicrobial activity of 45 extracts of medicinal plants were tested on Xanthomonas campestris pv. vesicatoria, Ralstonia solanacearum and Clavibacter michiganense subsp. michiganense. Some assays were done to verify the capability of these plants extrats to show an antibiosis. Five extracts (EAFQ, SM1, SM12, SM16, SA1) shown positive activity. The extract EAFQ expressed bactericide activity on C. michiganense. It suggests the possibility of using these actives substances in natura or as a model to synthesize industrialized products, intendind field utilization.

Modulation of the pharmacological effects of enzymatically-active PLA 2 by BTL-2, an isolectin isolated from the Bryothamnion triquetrum red alga

Background. An interaction between lectins from marine algae and PLA 2 from rattlesnake was suggested some years ago. We, herein, studied the effects elicited by a small isolectin (BTL-2), isolated from Bryothamnion triquetrum, on the pharmacological and biological activities of a PLA 2 isolated from rattlesnake venom (Crotalus durissus cascavella), to better understand the enzymatic and pharmacological mechanisms of the PLA 2 and its complex. Results. This PLA2 consisted of 122 amino acids (approximate molecular mass of 14 kDa), its pI was estimated to be 8.3, and its amino acid sequence shared a high degree of similarity with that of other neurotoxic and enzymatically-active PLA2s. BTL-2 had a molecular mass estimated in approximately 9 kDa and was characterized as a basic protein. In addition, BTL-2 did not exhibit any enzymatic activity. The PLA2 and BTL-2 formed a stable heterodimer with a molecular mass of approximately 24-26 kDa, estimated by molecular exclusion HPLC. In the presence of BTL-2, we observed a significant increase in PLA2 activity, 23% higher than that of PLA2 alone. BTL-2 demonstrated an inhibition of 98% in the growth of the Gram-positive bacterial strain, Clavibacter michiganensis michiganensis (Cmm), but only 9.8% inhibition of the Gram-negative bacterial strain, Xanthomonas axonopodis pv passiflorae (Xap). PLA2 decreased bacterial growth by 27.3% and 98.5% for Xap and Cmm, respectively, while incubating these two proteins with PLA2-BTL-2 inhibited their growths by 36.2% for Xap and 98.5% for Cmm. PLA2 significantly induced platelet aggregation in washed platelets, whereas BTL-2 did not induce significant platelet aggregation in any assay. However, BTL-2 significantly inhibited platelet aggregation induced by PLA2. In addition, PLA 2 exhibited strong oedematogenic activity, which was decreased in the presence of BTL-2. BTL-2 alone did not induce oedema and did not decrease or abolish the oedema induced by the 48/80 compound. Conclusion. The unexpected results observed for the PLA2-BTL-2 complex strongly suggest that the pharmacological activity of this PLA2 is not solely dependent on the presence of enzymatic activity, and that other pharmacological regions may also be involved. In addition, we describe for the first time an interaction between two different molecules, which form a stable complex with significant changes in their original biological action. This opens new possibilities for understanding the function and action of crude venom, an extremely complex mixture of different molecules. © 2008 Oliveira et al; licensee BioMed Central Ltd.