In planta proteomics and proteogenomics of the biotrophic barley fungal pathogen blumeria graminis f. sp hordei

To further our understanding of powdery mildew biology during infection, we undertook a systematic shotgun proteomics analysis of the obligate biotroph Blumeria graminis f. sp. hordei at different stages of development in the host. Moreover we used a proteogenomics approach to feed information into the annotation of the newly sequenced genome. We analyzed and compared the proteomes from three stages of development representing different functions during the plant-dependent vegetative life cycle of this fungus. We identified 441 proteins in ungerminated spores, 775 proteins in epiphytic sporulating hyphae, and 47 proteins from haustoria inside barley leaf epidermal cells and used the data to aid annotation of the B. graminis f. sp. hordei genome. We also compared the differences in the protein complement of these key stages. Although confirming some of the previously reported findings and models derived from the analysis of transcriptome dynamics, our results also suggest that the intracellular haustoria are subject to stress possibly as a result of the plant defense strategy, including the production of reactive oxygen species. In addition, a number of small haustorial proteins with a predicted N-terminal signal peptide for secretion were identified in infected tissues: these represent candidate effector proteins that may play a role in controlling host metabolism and immunity. Molecular & Cellular Proteomics 8: 2368-2381, 2009.

In planta proteomics and proteogenomics of the biotrophic barley fungal pathogen blumeria graminis f. sp hordei

To further our understanding of powdery mildew biology during infection, we undertook a systematic shotgun proteomics analysis of the obligate biotroph Blumeria graminis f. sp. hordei at different stages of development in the host. Moreover we used a proteogenomics approach to feed information into the annotation of the newly sequenced genome. We analyzed and compared the proteomes from three stages of development representing different functions during the plant-dependent vegetative life cycle of this fungus. We identified 441 proteins in ungerminated spores, 775 proteins in epiphytic sporulating hyphae, and 47 proteins from haustoria inside barley leaf epidermal cells and used the data to aid annotation of the B. graminis f. sp. hordei genome. We also compared the differences in the protein complement of these key stages. Although confirming some of the previously reported findings and models derived from the analysis of transcriptome dynamics, our results also suggest that the intracellular haustoria are subject to stress possibly as a result of the plant defense strategy, including the production of reactive oxygen species. In addition, a number of small haustorial proteins with a predicted N-terminal signal peptide for secretion were identified in infected tissues: these represent candidate effector proteins that may play a role in controlling host metabolism and immunity. Molecular & Cellular Proteomics 8: 2368-2381, 2009.

Proteogenomics and in silico structural and functional annotation of the barley powdery mildew Blumeria graminis f. sp. hordei

Blumeria graminis is an economically important obligate plant-pathogenic fungus, whose entire genome was recently sequenced and manually annotated using ab initio in silico predictions [7]. Employing large scale proteogenomic analysis we are now able to verify independently the existence of proteins predicted by 24% of open reading frame models. We compared the haustoria and sporulating hyphae proteomes and identified 71 proteins exclusively in haustoria, the feeding and effector-delivery organs of the pathogen. These proteins are ‘significantly smaller than the rest of the protein pool and predicted to be secreted. Most do not share any similarities with Swiss–Prot or Trembl entries nor possess any identifiable Pfam domains. We used a novel automated prediction pipeline to model the 3D structures of the proteins, identify putative ligand binding sites and predict regions of intrinsic disorder. This revealed that the protein set found exclusively in haustoria is significantly less disordered than the rest of the identified Blumeria proteins or random (and representative) protein sets generated from the yeast proteome. For most of the haustorial proteins with unknown functions no good templates could be found, from which to generate high quality models. Thus, these unknown proteins present potentially new protein folds that can be specific to the interaction of the pathogen with its host.

Biologia de Psyllobora confluens alimentada com o fungo Erysiphe cichoracearum

O objetivo deste trabalho foi avaliar os aspectos biológicos de Psyllobora confluens alimentada com o fungo Erysiphe cichoracearum em folhas de quiabeiro. Larvas recém-eclodidas de P. confluens foram individualizadas e alimentadas diariamente com hifas e conídios do fungo, em folhas de quiabeiro, a 23,6±3 e 27,4±2ºC e fotófase de 12 horas. A duração dos estágios imaturos, os aspectos reprodutivos e a longevidade de P. confluens foram avaliados. O período de desenvolvimento de larva a adulto foi 20,5 dias a 23,6ºC e 16,6 dias a 27,4ºC. em ambas as temperaturas, 80% das larvas de P. confluens atingiram a fase adulta. Fêmeas de P. confluens apresentaram período de oviposição de 32,7 dias, com capacidade diária e total de oviposição de 16,8 e 439,9 ovos, respectivamente. A longevidade das fêmeas foi de 46,1 dias e a dos machos 58,7 dias. P. confluens, ao se alimentar do fungo E. cichoracearum em folhas de quiabeiro, completa o ciclo de vida e apresenta elevados padrões de sobrevivência e reprodução.

Caracterização histoquimica de folhas de mentas infectadas por Puccinia menthae e Erysiphe biocellata

Pós-graduação em Agronomia (Horticultura) - FCA

Reação de linhagens de ervilha de vagens comestíveis (Pisum sativum L.) ao oídio (Erysiphe pisi DC.)

Pós-graduação em Agronomia (Proteção de Plantas) - FCA

Avaliação de germoplasma de quiabeiro (Abelmoschus esculentus) quanto à resistência ao oídio (Erysiphe cichoracearum)

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Ocorrência de Meloidogyne graminis em grama no estado de São Paulo

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Identifying races of Puccinia graminis f. sp. avenae; a modified international system

Bibliography: p. 19-23.

Identification of a major locus conferring resistance to powdery mildew (Erysiphe polygoni DC) in mungbean (Vigna radiata L. Wilczek) by QTL analysis

A major locus conferring resistance to the causal organism of powdery mildew, Erysiphe polygoni DC,, in mungbean (Vigna radiata L. Wilczek) was identified using QTL analysis with a population of 147 recombinant inbred individuals. The population was derived from a cross between 'Berken', a highly susceptible variety, and ATF 3640, a highly resistant line. To test for response to powdery mildew, F-7 and F-8 lines were inoculated by dispersing decaying mungbean leaves with residual conidia of E. polygoni amongst the young plants to create an artificial epidemic and assayed in a glasshouse facility. To generate a linkage map, 322 RFLP clones were tested against the two parents and 51 of these were selected to screen the mapping population. The 51 probes generated 52 mapped loci, which were used to construct a linkage map spanning 350 cM of the mungbean genome over 10 linkage groups. Using these markers, a single locus was identified that explained up to a maximum of 86% of the total variation in the resistance response to the pathogen.

Influence of light and leaf epicuticular wax layer on Phakopsora pachyrhizi infection in soybean

Influence of light and leaf epicuticular wax layer on Phakopsora pachyrhizi infection in soybean Asian rust, caused by the fungus Phakopsora pachyrhizi, is one of the most serious phytosanitary problems of soybean in Brazil, especially because no cultivars with satisfactory resistance levels as yet exist. The objective of this study was to evaluate the influence of luminosity and of leaf epicuticular wax on the infection of soybean by P. pachyrhizi. The adaxial and abaxial leaflet surfaces of the first trifoliate leaf from cultivar BRS 154, phenological stage V2, were inoculated with a suspension of 105 uredospores/mL. The plants were kept for 24 hours in a humid chamber at temperature of 23 degrees C, in light or dark conditions, using a factorial design. Subsequently, the plants were maintained for 14 days under a 12-hour photoperiod. The disease severity and density were evaluated. For in vitro experiments, in light or dark conditions, the evaluation was done in terms of uredospore germination and appressorium formation. The wax content of adaxial and abaxial leaflets was analyzed quantitatively using chloroform extraction and ultrastructurally using scanning electron microscope. Higher density and severity were observed when the adaxial surface was inoculated, with later incubation of the plants in the dark, with no significant interaction between these factors. Spore germination in the dark (40.7%) was statistically different from spore germination in the light (28.5%). The same effect was observed with appressorium formation, in the dark (24.7%) and in the light (12.8%). The quantity and the ultrastructural aspects of epicuticular wax content did not show differences between the adaxial and abaxial surfaces; nor did they show any effect on infection by Phakopsora pachyrhizi in the soybean cultivar studied.

Desempenho de genótipos de feijão-mungo-verde semeados no inverno na Zona da Mata de Minas Gerais

Avaliou-se o desempenho do feijão-mungo-verde, semeado no inverno, na Zona da Mata de Minas Gerais, a 400 (Oratórios) e 720 m (Coimbra) de altitude. Dez genótipos foram semeados no final de julho, ou no início de agosto, no delineamento em blocos ao acaso, com quatro repetições. Irrigações por aspersão complementaram as chuvas. A primeira vagem amadureceu aos 73,3 (Oratórios) e aos 79,9 dias após a emergência (DAE) (Coimbra). A altura das plantas foi maior em Oratórios (62,4 cm) do que em Coimbra (37,1 cm). Em Coimbra, doenças causadas por Erysiphe polygoni e Ascochita sp. foram moderadas. Em Oratórios, foram feitas três colheitas entre 81 e 94 DAE; em Coimbra, duas: 88 e 100 DAE. As produtividades médias foram de 1093 (Oratórios) e 801 kg/ha (Coimbra). As sementes colhidas em Oratórios apresentaram aspecto ótimo ou bom; em Coimbra, bom ou regular. Concluímos que o plantio do feijão-mungo-verde no inverno, em altitudes entre 400 e 720 m, proporciona produtividades relativamente modestas, especialmente a 720 m, onde as condições climáticas favorecem algumas doenças.

The grapevine defences

Grapevine is one of the most cultivated fruit crop worldwide with Vitis vinifera being the species with the highest economical importance due to the high quality standards of its berries. Nonetheless, it is also the most susceptible Vitis species to fungal pathogens. Among others, relevant fungal diseases currently threatening grapevine cultures are powdery mildew, caused by Erysiphe necator, and esca, a disease complex comprised of several fungi in which Phaeomoniella chlamydospora and Phaeoacremonium aleophilum participate.(...)

Analysis of epidermis- and mesophyll-specific transcript accumulation in powdery mildew-inoculated wheat leaves.

Powdery mildew is an important disease of wheat caused by the obligate biotrophic fungus Blumeria graminis f. sp. tritici. This pathogen invades exclusively epidermal cells after penetrating directly through the cell wall. Because powdery mildew colonizes exclusively epidermal cells, it is of importance not only to identify genes which are activated, but also to monitor tissue specificity of gene activation. Acquired resistance of wheat to powdery mildew can be induced by a previous inoculation with the non-host pathogen B. graminis f. sp. hordei, the causal agent of barley powdery mildew. The establishment of the resistant state is accompanied by the activation of genes. Here we report the tissue-specific cDNA-AFLP analysis and cloning of transcripts accumulating 6 and 24 h after the resistance-inducing inoculation with B. graminis f. sp. hordei. A total of 25,000 fragments estimated to represent about 17,000 transcripts were displayed. Out of these, 141 transcripts, were found to accumulate after Bgh inoculation using microarray hybridization analysis. Forty-four accumulated predominantly in the epidermis whereas 76 transcripts accumulated mostly in mesophyll tissue.