Levantamento de Diaphorina citri no estado do Pará.

Considerando a importância de D. citri na disseminação do HLB, este trabalho teve como objetivo o monitoramento do psilídeo em plantas de citros e principalmente de murta em municípios do estado do Pará

Métodos para detecção de Clavibacter michiganense subsp. michiganense e Xanthomonas campestris pv. vesicatoria em sementes de tomate.

Visando detectar Clavibacter michiganense subsp. michiganense (Cm) e Xanthomonas campestris pv. vesicatoria (Xcv) em sementes de tomate, duas técnicas foram comparadas: meio semi-seletivo e planta indicadora. Os seguintes parâmetros foram avaliados: soluções extratoras de Cm e Xcv de sementes inteiras e moídas, especificidade e sensibilidade. Os resultados mostraram que os meios semi-seletivos MB1M (MB1 + telurito de potássio, ácido borico e benomil) e TAM (peptona, brometo de potássio, cloreto de cálcio, agar + Tween 80, cefalexina e clorotalonil), foram mais eficientes para detecção de Cm e Xcv, a partir de sementes moídas em tampão fosfato do que os meios disponíveis e, apresentaram maior especificidade e sensibilidade, detectando 10(2) - 10(3) ufc/ml de Cme Xcv em comparacao a 10(3) - 10(4) ufc/ml da inoculação em plântulas de tomateiro (cvs. Angela Gigante e Santa Cruz).

Laterally transferred genomic islands in Xanthomonadales related to pathogenicity and primary metabolism

Lateral gene transfer (LGT) is considered as one of the drivers in bacterial genome evolution, usually associated with increased fitness and/or changes in behavior, especially if one considers pathogenic vs. non-pathogenic bacterial groups. The genomes of two phytopathogens, Xanthomonas campestris pv. campestris and Xanthomonas axonopodis pv. citri, were previously inspected for genome islands originating from LGT events, and, in this work, potentially early and late LGT events were identified according to their altered nucleotide composition. The biological role of the islands was also assessed, and pathogenicity, virulence and secondary metabolism pathways were functions highly represented, especially in islands that were found to be recently transferred. However, old islands are composed of a high proportion of genes related to cell primary metabolic functions. These old islands, normally undetected by traditional atypical composition analysis, but confirmed as product of LGT by atypical phylogenetic reconstruction, reveal the role of LGT events by replacing core metabolic genes normally inherited by vertical processes.

PILZ Protein Structure and Interactions with PILB and the FIMX EAL Domain: Implications for Control of Type IV Pilus Biogenesis

The PilZ protein was originally identified as necessary for type IV pilus (T4P) biogenesis. Since then, a large and diverse family of bacterial PilZ homology domains have been identified, some of which have been implicated in signaling pathways that control important processes, including motility, virulence and biofilm formation. Furthermore, many PilZ homology domains, though not PilZ itself, have been shown to bind the important bacterial second messenger bis(3`-> 5`)cyclic diGMP (c-diGMP). The crystal structures of the PilZ orthologs from Xanthomonas axonopodis pv Citri (PilZ(XAC1133), this work) and from Xanthomonas campestris pv campestris (XC1028) present significant structural differences to other PilZ homologs that explain its failure to bind c-diGMP. NMR analysis of PilZ(XAC1133) shows that these structural differences are maintained in solution. In spite of their emerging importance in bacterial signaling, the means by which NZ proteins regulate specific processes is not clear. In this study, we show that PilZ(XAC1133) binds to PilB, an ATPase required for TV polymerization, and to the EAL domain of FiMX(XAC2398), which regulates TV biogenesis and localization in other bacterial species. These interactions were confirmed in NMR, two-hybrid and far-Western blot assays and are the first interactions observed between any PilZ domain and a target protein. While we were unable to detect phosphodiesterase activity for FimXX(AC2398) in vitro, we show that it binds c-diGMP both in the presence and in the absence of PilZ(XAC1133). Site-directed mutagenesis studies for conserved and exposed residues suggest that PilZ(XAC1133) interactions with FimX(XAC2398) and PilB(XAC3239) are mediated through a hydrophobic surface and an unstructured C-terminal extension conserved only in PilZ orthologs. The FimX-PilZ-PilB interactions involve a full set of ""degenerate"" GGDEF, EAL and PilZ domains and provide the first evidence of the means by which PilZ orthologs and FimX interact directly with the TP4 machinery. (C) 2009 Elsevier Ltd. All rights reserved.

Cancro cítrico em viveiros submetidos a manejos convencional e orgânico

O objetivo deste trabalho foi o de avaliar o desenvolvimento de portaenxertos de citros Poncirus trifoliata, índices de ataque de cancro cítrico causado por Xanthomonas axonopodis pv. citri e controle dessa doença com pulverizações cúpricas em dois viveiros, um convencional e outro orgânico, artificialmente inoculados, no Centro de Formação da EMATER, situado no município de Montenegro/RS, no Estado do Rio Grande do Sul. Para controle do cancro cítrico foram testadas pulverizações cúpricas em diferentes concentrações e freqüências utilizando-se calda bordalesa no viveiro orgânico e oxicloreto de cobre no viveiro convencional. Foram avaliados: o crescimento do diâmetro do caule dos portaenxertos; a produção de matéria seca da parte aérea e a contagem do número de lesões de cancro cítrico presentes em folhas e ramos. Com os dados obtidos foi possível verificar que os tratamentos cúpricos não controlaram o cancro cítrico; ambos os viveiros, convencional e orgânico proporcionaram desenvolvimento semelhante aos porta-enxertos; e, com pequenas variações, o cancro cítrico se desenvolveu com igual intensidade nos dois viveiros.

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.

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.

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.

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.

Optimal clone identifier based on dynamic rules and process algebra

The rule creation to clone selection in different projects is a hard task to perform by using traditional implementations to control all the processes of the system. The use of an algebraic language is an alternative approach to manage all of system flow in a flexible way. In order to increase the power of versatility and consistency in defining the rules for optimal clone selection, this paper presents the software OCI 2 in which uses process algebra in the flow behavior of the system. OCI 2, controlled by an algebraic approach was applied in the rules elaboration for clone selection containing unique genes in the partial genome of the bacterium Bradyrhizobium elkanii Semia 587 and in the whole genome of the bacterium Xanthomonas axonopodis pv. citri. Copyright© (2009) by the International Society for Research in Science and Technology.

Daños causados por cancro cítrico en la producción de naranja dulce 'Valencia'

The citrus canker, caused by Xanthomonas axonopodis pv. citri, is one of the main citrus diseases and has threatened the world's citriculture. In this study, the damage caused by citrus canker was quantified in a sweet orange culture, cv. Valencia [Citrus sinensis (L.) Osb.], grafted over 15 year-old Rangpur lime (C. limonia Osb.), in the municipality of Guatambu, SC, Brazil, in the crop of 2004/05. Evaluations of the number of fruits per plant (NF), fruit yield per plant (P), citrus canker incidence on the fruit (IFR), and incidence (IL) and severity (SV) of citrus canker on 80 leaves per orange tree were performed in the harvest period, applying diagrammatical scales. Plants had large variation in SV (0.16%-1.09%), IL (12.50%-56.25%) and IFR (12.30%-56.09%). Linear regressions were significant between IL x SV, NF x SV, P x IFR, and NF x IFR. There was no significant difference (p>0.05) between the mean estimated values of IL in different quadrants of the crowns of the orange trees (N, S, L and O). Each 1% increase in IFR reduced 2.16 kg and 21.3 fruits per tree.

Mapas de zonas de risco de epidemias para doenças dos citros no Estado de São Paulo

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Development of PCR-based markers for detection of Leifsonia xyli subsp. xyli in fibrovascular fluid of infected sugarcane plants

DNA of Leifsonia xyli subsp. xyli (Lxx), the causal agent of ratoon stunting disease of sugarcane, was detected in the fibrovascular fluid of sugarcane plants using random amplified polymorphic DNA PCR-based amplification using two 10-mer oligonucleotide primers. The primers OPC-02 and OPC-11 produced Lxx-specific markers of approximately 800 bp and 1000 bp, respectively. A cloned DNA fragment from the 800 bp PCR product (pSKC2-800) hybridised to a single genomic DNA fragment from Lxx when used as a probe in Southern hybridisation. This cloned fragment did not hybridise to L. xyli subsp. cynodontis (Lxc), or L. xyli-like bacteria isolated from grasses in Australia, indicating the usefulness of this DNA fragment as a specific probe for Lxx. A cloned fragment from the 1000 bp PCR product ( pSKC11-1000) hybridised to three genomic fragments in Lxx isolates, one genomic fragment in two of the four isolates of L. xyli-like bacteria, and in two of the four isolates of Lxc isolated from the USA. These results indicate that L. xyli-like bacteria are more likely to be related to Lxc than Lxx. These probes did not hybridise to the DNA from strains of the species of Clavibacter, Rathayibacter, Acidovorax, Ralstonia, Pseudomonas and Xanthomonas tested. Two oligonucleotide primers (21-mer) designed from the pSKC2-800 sequences specifically amplified template DNA from Lxx and detected as few as 5 x 10(4) cells/mL in fibrovascular fluid from sugarcane plants infected with Lxx.

Genetic uniformity of international isolates of Leifsonia xyli subsp xyli, causal agent of ratoon stunting disease of sugarcane

An international collection of the sugarcane ratoon stunting disease pathogen, Leifsonia xyli subsp. xyli, was analysed to assess genetic diversity. DNA fingerprinting using BOX primers was performed on 105 isolates, comprising 65 Australian isolates and an additional 40 isolates from Indonesia (n = 8), Japan (n = 1), USA (n = 3), Brazil (n = 2), Mali (n = 2), Zimbabwe (n = 13), South Africa (n = 9) and Reunion (n = 2). Sixty-two of these isolates were also screened using ERIC primers. No variation was found among any of the isolates. The intergenic spacer (IGS) region of the ribosomal RNA genes from 54 isolates was screened for sequence variation using single-stranded conformational polymorphism (SSCP), but none was observed. Direct sequencing of the IGS from a subset of nine isolates, representing all of the countries sampled in this study, confirmed the results of the SSCP analysis. Likewise, no sequence variation was found in the 16S ribosomal RNA genes of the same subset. Four Colombian isolates from sugarcane, morphologically similar to L. xyli subsp. xyli, were putatively shown to be an undescribed Agrococcus species of unknown pathogenicity. The lack of genetic variation among L. xyli subsp. xyli isolates, independent of time of sampling, cultivar of isolation, or country of origin, suggests the worldwide spread of a single pathogenic clone, and further suggests that sugarcane cultivars resistant to ratoon stunting disease in one area should retain this property in other regions.

DrsG from Streptococcus dysgalactiae subsp equisimilis inhibits the antimicrobial peptide LL- 37

SIC and DRS are related proteins present in only four of the more than 200 Streptococcus pyogenes emm-types. These proteins inhibit complement mediated lysis and/or the activity of certain antimicrobial peptides. A gene encoding a homologue of these proteins, herein called DrsG, has been identified in the related bacterium Streptococcus dysgalactiae subsp equisimilis (SDSE). Here we show that geographically dispersed isolates representing 14 of 50 emm-types examined possess variants of drsG. However not all isolates within the drsG-positive emm-types possess the gene. Sequence comparisons also reveal a high degree of conservation in different SDSE emm-types. To examine the biological activity of DrsG, recombinant versions of two major DrsG variants, DrsGS and DrsGL, were expressed and purified. Western blot analysis using antisera raised to these proteins demonstrated both variants to be expressed and secreted into culture supernatant. Unlike SIC, but similar to DRS, DrsG does not inhibit complement mediated lysis. However, like both SIC and DRS, DrsG is a ligand of the cathelcidin LL-37 and is inhibitory to its bactericidal activity in in vitro assays. The greatest similarity between DrsG and DRS/SIC is found in the signal sequence at the amino terminus and proline rich domains in the C-terminal half of the protein. Conservation of prolines in this latter region also suggests these residues are important in the biology of this family of proteins. This is the first report demonstrating the activity of an AMP inhibitory protein in SDSE. These results also suggest that inhibition of AMP activity is the primary function of this family of proteins. The acquisition of complement inhibitory activity of SIC may reflect its continuing evolution.