28 results on '"Rincones, Johana"'
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2. In Vitro Production of Biotrophic-Like Cultures of Crinipellis perniciosa, the Causal Agent of Witches’ Broom Disease of Theobroma cacao
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Meinhardt, Lyndel W., Bellato, Cláudia de M., Rincones, Johana, Azevedo, Ricardo A., Cascardo, Julio C. M., and Pereira, Gonçalo A. G.
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- 2006
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3. Bacteria modulate the degree of amphimix of their symbiotic entomopathogenic nematodes (Heterohabditis spp) in response to nutritional stress
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Rincones, Johana, Mauléon, Hervé, and Jaffe, Klaus
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- 2001
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4. The genome sequence of Propionibacterium acidipropionici provides insights into its biotechnological and industrial potential
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Parizzi Lucas P, Grassi Maria Carolina B, Llerena Luige A, Carazzolle Marcelo F, Queiroz Verônica L, Lunardi Inês, Zeidler Ane F, Teixeira Paulo JPL, Mieczkowski Piotr, Rincones Johana, and Pereira Gonçalo AG
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Genome ,Propionibacterium acidipropionici ,Propionic acid ,Biotechnology ,TP248.13-248.65 ,Genetics ,QH426-470 - Abstract
Abstract Background Synthetic biology allows the development of new biochemical pathways for the production of chemicals from renewable sources. One major challenge is the identification of suitable microorganisms to hold these pathways with sufficient robustness and high yield. In this work we analyzed the genome of the propionic acid producer Actinobacteria Propionibacterium acidipropionici (ATCC 4875). Results The assembled P. acidipropionici genome has 3,656,170 base pairs (bp) with 68.8% G + C content and a low-copy plasmid of 6,868 bp. We identified 3,336 protein coding genes, approximately 1000 more than P. freudenreichii and P. acnes, with an increase in the number of genes putatively involved in maintenance of genome integrity, as well as the presence of an invertase and genes putatively involved in carbon catabolite repression. In addition, we made an experimental confirmation of the ability of P. acidipropionici to fix CO2, but no phosphoenolpyruvate carboxylase coding gene was found in the genome. Instead, we identified the pyruvate carboxylase gene and confirmed the presence of the corresponding enzyme in proteome analysis as a potential candidate for this activity. Similarly, the phosphate acetyltransferase and acetate kinase genes, which are considered responsible for acetate formation, were not present in the genome. In P. acidipropionici, a similar function seems to be performed by an ADP forming acetate-CoA ligase gene and its corresponding enzyme was confirmed in the proteome analysis. Conclusions Our data shows that P. acidipropionici has several of the desired features that are required to become a platform for the production of chemical commodities: multiple pathways for efficient feedstock utilization, ability to fix CO2, robustness, and efficient production of propionic acid, a potential precursor for valuable 3-carbon compounds.
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- 2012
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5. A genome survey of Moniliophthora perniciosa gives new insights into Witches' Broom Disease of cacao
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Bailey Bryan A, Guiltinan Mark J, Barbosa Luciana V, Neto Aristóteles, Neto José, Gonçalves Marilda S, Gramacho Karina P, Castro Luis AB, de Moraes Marcos H, Araújo Marcos, Rio Maria, Pires Acássia BL, de Oliveira Bruno V, Thomazella Daniela PT, García Odalys, Estrela Raíssa C, Vidal Ramon O, Carrer Helaine, Cunha Anderson F, Carraro Dirce M, Cotomacci Carolina, Rincones Johana, Parizzi Lucas P, Formighieri Eduardo F, Costa Gustavo GL, Carazzolle Marcelo F, Mondego Jorge MC, Meinhardt Lyndel W, Cascardo Julio CM, and Pereira Gonçalo AG
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Biotechnology ,TP248.13-248.65 ,Genetics ,QH426-470 - Abstract
Abstract Background The basidiomycete fungus Moniliophthora perniciosa is the causal agent of Witches' Broom Disease (WBD) in cacao (Theobroma cacao). It is a hemibiotrophic pathogen that colonizes the apoplast of cacao's meristematic tissues as a biotrophic pathogen, switching to a saprotrophic lifestyle during later stages of infection. M. perniciosa, together with the related species M. roreri, are pathogens of aerial parts of the plant, an uncommon characteristic in the order Agaricales. A genome survey (1.9× coverage) of M. perniciosa was analyzed to evaluate the overall gene content of this phytopathogen. Results Genes encoding proteins involved in retrotransposition, reactive oxygen species (ROS) resistance, drug efflux transport and cell wall degradation were identified. The great number of genes encoding cytochrome P450 monooxygenases (1.15% of gene models) indicates that M. perniciosa has a great potential for detoxification, production of toxins and hormones; which may confer a high adaptive ability to the fungus. We have also discovered new genes encoding putative secreted polypeptides rich in cysteine, as well as genes related to methylotrophy and plant hormone biosynthesis (gibberellin and auxin). Analysis of gene families indicated that M. perniciosa have similar amounts of carboxylesterases and repertoires of plant cell wall degrading enzymes as other hemibiotrophic fungi. In addition, an approach for normalization of gene family data using incomplete genome data was developed and applied in M. perniciosa genome survey. Conclusion This genome survey gives an overview of the M. perniciosa genome, and reveals that a significant portion is involved in stress adaptation and plant necrosis, two necessary characteristics for a hemibiotrophic fungus to fulfill its infection cycle. Our analysis provides new evidence revealing potential adaptive traits that may play major roles in the mechanisms of pathogenicity in the M. perniciosa/cacao pathosystem.
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- 2008
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6. Apoplastic and intracellular plant sugars regulate developmental transitions in witches' broom disease of cacao
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Barau, Joan Grande, 1981, Carvalho, Vinicius Miessler de Andrade, 1989, Teixeira, Gleidson Silva, 1984, Zaparoli, Gustavo Henrique Alcalá, 1983, Rio, Maria Carolina Scatolin do, 1977, Rincones, Johana, 1977, Pereira, Gonçalo Amarante Guimarães, 1964, and UNIVERSIDADE ESTADUAL DE CAMPINAS
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Moniliophthora perniciosa ,Cacao ,Starvation ,Autophagy ,Autofagia ,Artigo original ,Cacaueiro ,Senescence ,Hemibiotrophic - Abstract
Agradecimentos: The authors are grateful for the assistance of Dr Lauro Tatsuo Kubota and Dr Jailson Cardoso Dias in the chromatographic detection and analysis of carbohydrates of cacao apoplastic fluid, and Marcelo Carazzolle, Osvaldo Reis Junior, and Gustavo Gilson Lacerda Costa for the bioinformatics support and management of databases consulted in the conduct of this work. This work was financially supported by CNPq (472710/2008-7) and FAPESP (2006/56942-0, 2009/17507-5, and 2009/50119-9). The authors declare no conflicts of interest Abstract: Witches’ broom disease (WBD) of cacao differs from other typical hemibiotrophic plant diseases by its unusually long biotrophic phase. Plant carbon sources have been proposed to regulate WBD developmental transitions; however, nothing is known about their availability at the plant–fungus interface, the apoplastic fluid of cacao. Data are provided supporting a role for the dynamics of soluble carbon in the apoplastic fluid in prompting the end of the biotrophic phase of infection. Carbon depletion and the consequent fungal sensing of starvation were identified as key signalling factors at the apoplast. MpNEP2, a fungal effector of host necrosis, was found to be up-regulated in an autophagic-like response to carbon starvation in vitro. In addition, the in vivo artificial manipulation of carbon availability in the apoplastic fluid considerably modulated both its expression and plant necrosis rate. Strikingly, infected cacao tissues accumulated intracellular hexoses, and showed stunted photosynthesis and the up-regulation of senescence markers immediately prior to the transition to the necrotrophic phase. These opposite findings of carbon depletion and accumulation in different host cell compartments are discussed within the frame of WBD development. A model is suggested to explain phase transition as a synergic outcome of fungal-related factors released upon sensing of extracellular carbon starvation, and an early senescence of infected tissues probably triggered by intracellular sugar accumulation CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - CNPQ FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESP Fechado
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- 2015
7. Apoplastic and intracellular plant sugars regulate developmental transitions in witches’ broom disease of cacao
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Barau, Joan, primary, Grandis, Adriana, additional, Carvalho, Vinicius Miessler de Andrade, additional, Teixeira, Gleidson Silva, additional, Zaparoli, Gustavo Henrique Alcalá, additional, do Rio, Maria Carolina Scatolin, additional, Rincones, Johana, additional, Buckeridge, Marcos Silveira, additional, and Pereira, Gonçalo Amarante Guimarães, additional
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- 2014
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8. A potential role for an extracellular methanol oxidase secreted by Moniliophthora perniciosa in Witches’ broom disease in cacao
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de Oliveira, Bruno V., primary, Teixeira, Gleidson S., additional, Reis, Osvaldo, additional, Barau, Joan G., additional, Teixeira, Paulo José P.L., additional, do Rio, Maria Carolina S., additional, Domingues, Romênia R., additional, Meinhardt, Lyndel W., additional, Paes Leme, Adriana F., additional, Rincones, Johana, additional, and Pereira, Gonçalo A.G., additional
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- 2012
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9. The hemibiotrophic cacao pathogen Moniliophthora perniciosa depends on a mitochondrial alternative oxidase for biotrophic development
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Thomazella, Daniela P. T., primary, Teixeira, Paulo José P. L., additional, Oliveira, Halley C., additional, Saviani, Elzira E., additional, Rincones, Johana, additional, Toni, Isabella M., additional, Reis, Osvaldo, additional, Garcia, Odalys, additional, Meinhardt, Lyndel W., additional, Salgado, Ione, additional, and Pereira, Gonçalo A. G., additional
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- 2012
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10. Spatial distribution of AT- and GC-rich DNA within interphase cell nuclei of Triatoma infestans Klug
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Alvarenga, Elenice M., primary, Mondin, Mateus, additional, Martins, James A., additional, Rodrigues, Vera L.C.C., additional, Vidal, Benedicto C., additional, Rincones, Johana, additional, Carazzolle, Marcelo F., additional, Andrade, Larissa Mara, additional, and Mello, Maria Luiza S., additional
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- 2011
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11. The Golden Bridge for Nature: The New Biology Applied to Bioplastics
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Rincones, Johana, primary, Zeidler, Ane F., additional, Grassi, Maria Carolina B., additional, Carazzolle, Marcelo F., additional, and Pereira, Gonçalo A. G., additional
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- 2009
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12. The glyceraldehyde-3-phosphate dehydrogenase gene of Moniliophthora perniciosa, the causal agent of witches' broom disease of Theobroma cacao
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Lima, Juliana O., primary, Pereira, Jorge F., additional, Rincones, Johana, additional, Barau, Joan G., additional, Araújo, Elza F., additional, Pereira, Gonçalo A.G., additional, and Queiroz, Marisa V., additional
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- 2009
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13. A genome survey of Moniliophthora perniciosa gives new insights into Witches' Broom Disease of cacao
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Mondego, Jorge MC, primary, Carazzolle, Marcelo F, additional, Costa, Gustavo GL, additional, Formighieri, Eduardo F, additional, Parizzi, Lucas P, additional, Rincones, Johana, additional, Cotomacci, Carolina, additional, Carraro, Dirce M, additional, Cunha, Anderson F, additional, Carrer, Helaine, additional, Vidal, Ramon O, additional, Estrela, Raíssa C, additional, García, Odalys, additional, Thomazella, Daniela PT, additional, de Oliveira, Bruno V, additional, Pires, Acássia BL, additional, Rio, Maria Carolina S, additional, Araújo, Marcos Renato R, additional, de Moraes, Marcos H, additional, Castro, Luis AB, additional, Gramacho, Karina P, additional, Gonçalves, Marilda S, additional, Neto, José P Moura, additional, Neto, Aristóteles Góes, additional, Barbosa, Luciana V, additional, Guiltinan, Mark J, additional, Bailey, Bryan A, additional, Meinhardt, Lyndel W, additional, Cascardo, Julio CM, additional, and Pereira, Gonçalo AG, additional
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- 2008
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14. The mitochondrial genome of the phytopathogenic basidiomycete Moniliophthora perniciosa is 109kb in size and contains a stable integrated plasmid
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Formighieri, Eduardo F., primary, Tiburcio, Ricardo A., additional, Armas, Eduardo D., additional, Medrano, Francisco J., additional, Shimo, Hugo, additional, Carels, Nicolas, additional, Góes-Neto, Aristóteles, additional, Cotomacci, Carolina, additional, Carazzolle, Marcelo F., additional, Sardinha-Pinto, Naiara, additional, Thomazella, Daniela P.T., additional, Rincones, Johana, additional, Digiampietri, Luciano, additional, Carraro, Dirce M., additional, Azeredo-Espin, Ana M., additional, Reis, Sérgio F., additional, Deckmann, Ana C., additional, Gramacho, Karina, additional, Gonçalves, Marilda S., additional, Moura Neto, José P., additional, Barbosa, Luciana V., additional, Meinhardt, Lyndel W., additional, Cascardo, Júlio C.M., additional, and Pereira, Gonçalo A.G., additional
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- 2008
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15. Moniliophthora perniciosa, the causal agent of witches’ broom disease of cacao: what's new from this old foe?
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MEINHARDT, LYNDEL W., primary, RINCONES, JOHANA, additional, BAILEY, BRYAN A., additional, AIME, M. CATHERINE, additional, GRIFFITH, GARETH W., additional, ZHANG, DAPENG, additional, and PEREIRA, GONÇALO A. G., additional
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- 2008
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16. Differential Gene Expression Between the Biotrophic-Like and Saprotrophic Mycelia of the Witches' Broom Pathogen Moniliophthora perniciosa
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Rincones, Johana, primary, Scarpari, Leandra M., additional, Carazzolle, Marcelo F., additional, Mondego, Jorge M. C., additional, Formighieri, Eduardo F., additional, Barau, Joan G., additional, Costa, Gustavo G. L., additional, Carraro, Dirce M., additional, Brentani, Helena P., additional, Vilas-Boas, Laurival A., additional, de Oliveira, Bruno V., additional, Sabha, Maricene, additional, Dias, Robson, additional, Cascardo, Júlio M., additional, Azevedo, Ricardo A., additional, Meinhardt, Lyndel W., additional, and Pereira, Gonçalo A. G., additional
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- 2008
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17. Characterization of necrosis and ethylene-inducing proteins (NEP) in the basidiomycete Moniliophthora perniciosa, the causal agent of witches' broom in Theobroma cacao
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Garcia, Odalys, primary, Macedo, Joci A.N., additional, Tibúrcio, Ricardo, additional, Zaparoli, Gustavo, additional, Rincones, Johana, additional, Bittencourt, Livia M.C., additional, Ceita, Geruza O., additional, Micheli, Fabienne, additional, Gesteira, Abelmon, additional, Mariano, Andréa C., additional, Schiavinato, Marlene A., additional, Medrano, Francisco J., additional, Meinhardt, Lyndel W., additional, Pereira, Gonçalo A.G., additional, and Cascardo, Júlio C.M., additional
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- 2007
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18. Genetic variability and chromosome-length polymorphisms of the witches' broom pathogen Crinipellis perniciosa from various plant hosts in South America
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RINCONES, Johana, primary, MAZOTTI, Gabriel D., additional, GRIFFITH, Gareth W., additional, POMELA, Alan, additional, FIGUEIRA, Antonio, additional, Leal, Gildemberg A., additional, QUEIROZ, Marisa V., additional, PEREIRA, Jorge F., additional, AZEVEDO, Ricardo A., additional, PEREIRA, Gonçalo A.G., additional, and MEINHARDT, Lyndel W., additional
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- 2006
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19. Electrophoretic karyotype analysis of Crinipellis perniciosa, the causal agent of witches' broom disease of Theobroma cacao
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Rincones, Johana, primary, Meinhardt, Lyndel W., additional, Vidal, Benedicto C., additional, and Pereira, Gonçalo A.G., additional
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- 2003
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20. Untitled.
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Rincones, Johana, Mauléon, Hervé, and Jaffe, Klaus
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Facultatively sexual entomopathogenic nematodes are a promising model for the experimental study of the adaptive values of sex. Our experiments in the laboratory showed that entomopathogenic nematodes display at least two different strategies in regulating the degree of amphimix as a response to nutritional stress. One strategy promotes the production of males, amphimix and the genetic variability of the offspring, improving the chances for a successful new adaptation. Another strategy increases the production of hermaphrodites at the expense of males, increasing the total number of reproductive individuals and thus the total number of offspring produced. Surprisingly, the strategy used depends upon the strain of symbiotic bacteria the nematodes are growing. The relevance of the results, in helping to discriminate between rival theories for the evolutionary maintenance of sex, is discussed. [ABSTRACT FROM AUTHOR]
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- 2001
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21. Rumo a uma plataforma iSUCCELL de Saccharomyces cerevisiae para a produção de etanol combustível
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Bermejo, Pamela Magalí, 1983, Gombert, Andreas Karoly, 1970, Stambuk, Boris Juan Carlos Ugarte, Rincones, Johana, Comelli, Raúl Nicolás, Basso, Thiago Olitta, Universidade Estadual de Campinas. Faculdade de Engenharia de Alimentos, Programa de Pós-Graduação em Bioenergia, and UNIVERSIDADE ESTADUAL DE CAMPINAS
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Sucrose ,Cellobiose ,Ethanol ,Etanol ,Celobiose ,Saccharomyces cerevisiae ,Biorrefinaria ,Sacarose ,Biorefinery - Abstract
Orientador: Andreas Karoly Gombert Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Engenharia de Alimentos Resumo: As atuais tecnologias de produção de etanol combustível de segunda geração (2G), com base em matérias-primas lignocelulósicas, enfrentam sérias dificuldades para se tornarem economicamente viáveis, tendo até agora alcançado volumes de produção bem abaixo das previsões feitas há uma ou duas décadas. Como iniciativa para enfrentar alguns dos desafios inerentes a esses processos industriais, aqui propomos um conceito alternativo, baseado na integração de tecnologias de primeira geração (1G) e 2G, no contexto de biorrefinarias de cana-de-açúcar. O conceito envolve uma mistura da corrente rica em sacarose do processo 1G com uma corrente 2G parcialmente hidrolisada, rica em celobiose, na etapa de fermentação. A estratégia também envolve o que chamamos de plataforma microbiana "iSUCCELL", responsável por realizar a fermentação da sacarose e da celobiose em etanol, via transporte ativo dos açúcares e suas subsequentes hidrólises intracelulares. Primeiramente, abordamos separadamente o metabolismo de cada dissacarídeo na levedura Saccharomyces cerevisiae. O componente "iCELL" foi estudado através da engenharia de linhagens de S. cerevisiae com o transportador de celobiose CDT-1 de Neurospora crassa e seis ?-glicosidases individuais de diferentes fungos filamentosos: uma de N. crassa (GH1-1), uma de Aspergillus niger (AN 8517) e quatro de Podospora anserina (PA 951, PA 3784, PA 6071 e PA 10293), sendo as cinco últimas enzimas ainda até então não descritas na literatura. Uma vez que nenhuma das linhagens engenheiradas inicialmente foi capaz de crescer em celobiose, uma posterior abordagem de evolução laboratorial foi conduzida para melhorar o desempenho cinético de todas as linhagens. Após a conclusão dos experimentos de evolução, embora a cinética de crescimento de todas as linhagens em celobiose como única fonte de carbono tenha melhorado, a análise dos dados de sequenciamento genômico revelou que pelo menos dois dos experimentos de evolução foram contaminados com Meyerozyma guilliermondii, uma espécie de levedura que conseguiu se estabelecer no sistema de cultivo, provavelmente devido à sua capacidade natural de utilizar a celobiose como fonte de carbono. Para o componente "iSUC" de nossa estratégia, lançamos mão das propriedades de uma linhagem iSUC de S. cerevisiae obtida em um trabalho anterior (Basso et al. 2011) e que apresenta maior rendimento em etanol, frente a uma linhagem referência. Avaliamos seu desempenho em situações mais condizentes com o cenário industrial, ou seja, em sistema de cultivo em batelada, com excesso de açúcar, e em um sistema miniaturizado que mimetiza o processo brasileiro de produção de etanol combustível 1G. Os resultados desses estudos mostraram que a linhagem iSUC não foi capaz de apresentar incremento no rendimento em etanol, quando comparada à linhagem referência que hidrolisa a sacarose predominantemente no meio extracelular. Analisados em conjunto, os resultados apresentados nesta tese, embora não tenham conduzido ao nosso objetivo inicial, abrem as portas para estudos futuros que visem demonstrar o potencial da abordagem iSUCCELL em uma linhagem industrial robusta e em um ambiente industrial real Abstract: Current second-generation (2G) fuel ethanol technologies are facing severe difficulties to become economically viable, having so far reached production volumes well below predictions made one or two decades ago. As an initiative to tackle some of the challenges inherent to these processes, here we propose an alternative concept that relies on the integration of first-generation (1G) and 2G sugarcane-based processes by mixing the sucrose-rich stream from the 1G process with a partially hydrolysed cellobiose-rich 2G stream in the fermentation step. The strategy also involves what we named a microbial "iSUCCELL" platform, responsible for carrying out fermentation of sucrose and cellobiose into ethanol, via active transport of the sugars and their subsequent intracellular hydrolyses. We first addressed the metabolism of each disaccharide in the yeast Saccharomyces cerevisiae, separately. The "iCELL" component was studied by engineering S. cerevisiae strains with the Neurospora crassa CDT-1 cellobiose transporter and six individual ?-glucosidases from different filamentous fungi: one from N. crassa (GH1-1), one from Aspergillus niger (AN 8517), and four from Podospora anserina (PA 951, PA 3784, PA 6071, and PA 10293), the latter five being hitherto undescribed enzymes. Since none of the engineered strains initially sustained growth on cellobiose, a further laboratory evolution approach was applied to improve the kinetic performances of all strains. After completion of the evolution experiments, although the growth kinetics of all strains on cellobiose as a sole carbon source indeed improved, analysis of whole-genome sequencing data revealed that at least two of the evolution experiments had been contaminated with Meyerozyma guilliermondii, a yeast species that managed to settle in the cell population, probably because of its natural capacity to utilise cellobiose as carbon source. For the "iSUC" component of our strategy, we leveraged the properties of a S. cerevisiae iSUC strain obtained in a previous work (Basso et al. 2011), in terms of increased ethanol yields vs. a reference strain, and evaluated its performance in situations which are closer to the industrial scenario, i.e., in batch operation with excess sugar and in a miniaturised system mimicking the Brazilian 1G fuel ethanol process. The results from these studies showed that the iSUC strain was not able to display any increment in the ethanol yield on substrate, when compared to the reference strain that mainly hydrolysed sucrose extracellularly. Analysed together, the results presented in this thesis, although not leading to reach our initial goal, open the door for future studies aiming at demonstrating the potential of the iSUCCELL approach in a robust industrial strain and in a real industrial environment Doutorado Bioenergia Doutora em Ciências CAPES 001 FAPESP
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- 2022
22. Genome and transcriptome analyzes of the fungal Ceratocystis cacaofunesta, aiming at the identification of possible targets for the control of Ceratocystis wilt disease
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Lopez Molano, Eddy Patricia, 1985, Pereira, Gonçalo Amarante Guimarães, 1964, Cabrera, Odalys Garcia, Mondego, Jorge Mauricio Costa, Rincones, Johana, Figueira, Antonio Vargas de Oliveira, Oliveira, Marcos Tulio de, Universidade Estadual de Campinas. Instituto de Biologia, Programa de Pós-Graduação em Genética e Biologia Molecular, and UNIVERSIDADE ESTADUAL DE CAMPINAS
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Cacao ,Ceratocystis cacaofunesta ,Fungos - Patogenicidade ,Fungi - Pathogenicity ,Genomes ,Cacaueiro ,Genomas - Abstract
Orientadores: Gonçalo Amarante Guimarães Pereira, Odalys Garcia Cabrera Tese (doutorado) - Universidade Estadual de Campinas, Instituto de Biologia Resumo: O cacaueiro (Theobroma cacao) é uma planta frutífera de importância econômica mundial, pois fornece matéria prima para diversos produtos, incluindo o chocolate. Porém, doenças fúngicas causam grandes perdas à cacauicultura. No Brasil e na América do sul, um dos principais patógenos do cacaueiro é o fungo ascomiceto Ceratocystis cacaofunesta, agente etiológico do Mal do Facão. Apesar do impacto econômico que esta doença causa nas plantações de cacau, ainda não existe uma estratégia efetiva e economicamente viável para o seu controle. Nesse contexto, o presente trabalho se propôs estudar a biologia do fungo focando principalmente o analises do genoma e transcriptoma de C. cacaofunesta com o objetivo de entender os mecanismos moleculares envolvidos na patogenicidade deste fungo e achar possíveis alvos para o controle desta importante doença. Para atingir este objetivo foram utilizadas diversas abordagens: (i) devido ao escasso conhecimento sobre a biologia do fungo, na primeira fase realizamos uma exaustiva revisão bibliográfica da historia e biologia do fungo e das outras espécies patogênicas que compreendem o gênero Ceratocystis, (capitulo publicado no livro Cacao Deseases). (ii) a segunda parte do trabalho consistiu no sequenciamento do genoma de C. cacaofunesta. Analisamos o conteúdo gênico do fungo e de outras espécies do gênero. Estas análises permitiram identificar grande variedade de proteínas codificadas no genoma e secretadas pelo fungo potencialmente relacionadas com o processo de patogenicidade. Dentre elas, destaca-se a expansão da família das fosfolipases C especificas para fosfatidil inositol (PI-PLC). A caracterização bioquímica e estrutural desta família de proteínas assim como estudos de genômica comparativa e discussão sobre o papel biológico destas proteínas no contexto do mal do facão foram objeto de uma publicação (BMC Genomics, Molano et al., 2018); (iii) o ultimo capitulo aborda estudos de resistência/susceptibilidade do fungo C. cacaofunesta a fungicidas cujos alvos são proteínas mitocondriais: Avaliação da respiração mitocondrial e ensaios de crescimento do fungo em presença de inibidores respiratórios mostraram que C. cacaofunesta tem resistência a altas concentrações do fungicida Azoxystrobina, inibidor de tipo estrobilurina que bloqueia a respiração via cadeia respiratória central (CRC), especificamente ocitocromo b da cadeia. Entretanto, o fungo é sensível a inibidores da via respiratória alternativa, especificamente inibidores da enzima oxidase alternativa (AOX). Foram analisados os efeitos destes inibidores no metabolismo geral de C. cacaofunesta mediante estudos de transcriptoma. Os resultados revelaram características até então inexploradas relacionadas à resposta do fungo a este tipo de drogas. As análises da resposta de C. cacaofunesta a inibidores da AOX sugerem que esta enzima é chave para a biologia e desenvolvimento do fungo tornando-a um alvo potencial para o controle deste fitopatógeno. Os resultados apresentados em cada uma das etapas deste trabalho auxiliaram no entendimento sobre a biologia do fungo C. cacaofunesta assim como a identificação de possíveis alvos para o controle da doença mal do facão. Adicionalmente, os resultados decorrentes deste projeto poderiam auxiliar no manejo de outras doenças de importância econômica causadas por espécies do gênero Ceratocystis Abstract: Cacao (Theobroma cacao) is a fruit tree of global economic importance, as it provides raw material for various products, including chocolate. However, fungal diseases cause great losses to cacao farming. In Brazil and South America, one of the main pathogens of cacao is the ascomycete fungus Ceratocystis cacaofunesta, an etiological agent of Mal do Facão. Despite the economic impact of this disease on cocoa plantations, there is still no effective and economically feasible strategy for its control. In this context, the present work aimed to study fungal biology focusing mainly on genome and transcriptome analysis of C. cacaofunesta in order to understand the molecular mechanisms involved in the pathogenicity of this fungus and to find possible targets for the control of this important disease. To achieve this objective, several approaches were used in this work: (i) due to the scarce knowledge about the biology of the fungus, in the first phase we performed an exhaustive bibliographic review of the history and biology of the fungus and the other pathogenic species that comprise the genus Ceratocystis, (chapter published in the book Cacao Deseases) the second part of the work consisted in sequencing the genome of C. cacaofunesta. We analyzed the gene content of the fungus and other species of the genus. These analyzes allowed to identify a large variety of proteins encoded in the genome and secreted by the fungus potentially related to the pathogenicity process. Among them, we highlight the expansion of the family of phospholipases C specific for phosphatidyl inositol (PI-PLC). The biochemical and structural characterization of this family of proteins as well as studies of comparative genomics and discussion about the biological role of these proteins in the context of macaw disease were published (BMC Genomics, Molano et al., 2018); (iii) The last chapter deals with studies of resistance / susceptibility of the fungus C. cacaofunesta to fungicides whose targets are mitochondrial proteins: Evaluation of mitochondrial respiration and growth tests of the fungus in the presence of respiratory inhibitors showed that C. cacaofunesta has resistance to high concentrations of the fungicide Azoxystrobin , a strobilurin-type inhibitor that blocks respiration via the central respiratory chain (CRC), specifically the ocytochrome b chain. However, the fungus is sensitive to alternative respiratory tract inhibitors, specifically the alternative oxidase enzyme (AOX) inhibitors. The effects of these inhibitors on the general metabolism of C. cacaofunesta were analyzed by transcriptome studies. The results revealed previously unexplored characteristics related to the fungus response to this type of drug. Analyzes of the response of C. cacaofunesta to AOX inhibitors suggest that this enzyme is key to the biology and development of the fungus making it a potential target for the control of this phytopathogen. The results presented in each of the stages of this work helped to understand the biology of the fungus C. cacaofunesta as well as the identification of possible targets for the control of the macaque disease. In addition, the results of this project could help in the management of other diseases of economic importance caused by species of the genus Ceratocystis Doutorado Genética de Microorganismos Doutora em Genética e Biologia Molecular CNPQ 142357/2014-6 CAPES 001
- Published
- 2019
23. Witche¿s broom disease : characterization of an extracellular methanol oxidase and of the secretome of Moniliophthora perniciosa
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Oliveira, Bruno Vaz de, 1985, Pereira, Gonçalo Amarante Guimarães, 1964, Rincones, Johana, Papes, Fabio, Cunha, Anderson Ferreira da, Rossi, Nilce Maria Martinez, Monteiro, Gisele, Universidade Estadual de Campinas. Instituto de Biologia, Programa de Pós-Graduação em Genética e Biologia Molecular, and UNIVERSIDADE ESTADUAL DE CAMPINAS
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Moniliophthora perniciosa ,Metanol como combustível ,Witche¿s broom disease ,Secretoma ,Vassoura-de-bruxa (Fitopatologia) ,Methanol oxidase ,Secretome - Abstract
Orientadores: Gonçalo Amarante Guimarães Pereira, Johana Rincones Pérez Tese (doutorado) - Universidade Estadual de Campinas, Instituto de Biologia Resumo: O fungo basidiomiceto Moniliophthora perniciosa é o agente causador da vassoura-de-bruxa do cacaueiro, doença que se tornou um dos principais problemas fitopatológicos do Brasil. O fungo possui duas fases de vida, biotrófica e necrotrófica, as quais estão associadas aos sintomas de vassoura-verde e vassoura-seca, respectivamente, durante a progressão da doença. Em vista dos grandes prejuízos econômicos causados pela vassoura-de-bruxa, foi iniciado o Projeto Genoma Vassoura-de-Bruxa, cujo objetivo é a decodificação genética do fungo com o intuito de identificar genes candidatos que possam estar relacionados ao estabelecimento e à progressão da doença para serem caracterizados. M. perniciosa é capaz de utilizar metanol como fonte única de carbono. Na maioria dos organismos estudados, a enzima metanol oxidase (MOX) é a responsável pelo metabolismo de metanol através do metabolismo peroxissomal, à qual também está associada uma catalase (CAT). Em plantas, uma das principais fontes de metanol é a pectina presente na estrutura da parede celular vegetal. Algumas espécies de fungos fitopatogênicos tem a capacidade de secretar oxalato como produto da enzima oxaloacetato acetilhidrolase (OAH), o qual é capaz de quelar íons cálcio da estrutura das pectinas, deixando-as vulneráveis ao ataque de enzimas que degradam a parede celular vegetal. Dentre essas enzimas, encontra-se a pectina metilesterase (PME), a qual é capaz de desmetilar as pectinas, liberando metanol. Como M. perniciosa possui em seu genoma sequências com similaridade a mox, cat, pme e oah, formulamos uma hipótese, segundo a qual M. perniciosa seria capaz de degradar o metanol proveniente das pectinas, através de um metabolismo peroxissomal, durante sua interação com o cacau na vassoura-de-bruxa. De fato, M. perniciosa é capaz de secretar oxalato, formando cristais de oxalato de cálcio. Nossos resultados sugerem fortemente que M. perniciosa utiliza o metanol proveniente das pectinas durante a vassoura-de-bruxa; no entanto, esse processo se daria através de um metabolismo extracelular, visto que M. perniciosa produz uma MOX e CAT extracelulares, e não através do metabolismo peroxissomal como inicialmente proposto. Além disso, M. perniciosa possui outras catalases, as quais se apresentam diferencialmente expressas nas diferentes fases do ciclo de vida do fungo e em diferentes estágios da progressão da vassoura-de-bruxa. Por fim, apresentamos uma análise preliminar do secretoma de M. perniciosa Abstract: The hemibiotrophic basidiomycete fungus Moniliophthora perniciosa is the causal agent of Witches' broom disease (WBD) in cacao. M. perniciosa is classified as a hemibiotrophic pathogen and presents two morphologically distinct life phases, biotrophic and necrotrophic, that are correlated with the green broom and dry broom symptoms, respectively, during the progression of WBD. The introduction of WBD in Bahia, the main Brazilian cacao-producing state, reduced the cacao production drastically and Brazil has become a net importer of cacao in order to supply the national chocolate industry. Due to the extreme losses in cacao production, the Witches' Broom Genome Project was initiated to decode the M. perniciosa genome and, based on the data acquired, select genes that could be relevant during the progression of WBD for characterization. M. perniciosa is able to grow in methanol as the sole carbon source. In methylotrophic yeasts and other methanol-degrading organisms, a methanol oxidase (MOX) is the key enzyme in methanol metabolism and it is also related to a catalase (CAT); both MOX and CAT characterize a peroxisomal metabolism. In plants, one of the main sources of methanol is the pectin present in the plant cell walls. Many phytopathogenic fungal species produce oxalate, a product of the enzyme oxaloacetate acetyl hydrolase (OAH), which removes calcium ions bound to pectin to produce calcium oxalate crystals, thus exposing the host cell walls to plant cell wall-degrading enzymes (PCWD) of fungal origin. One of the main PCWDs is pectin methylesterase (PME), an enzyme that removes the methyl ester radicals from esterified pectin, releasing methanol. As M. perniciosa possesses mox, cat, oah and pme on its genome, we hypothesized that M. perniciosa would utilize a peroxisomal metabolism to metabolize the methanol released from the pectin demethylation in WBD. Indeed, M. perniciosa secretes oxalate, thus producing calcium oxalate crystals. Our data strongly suggest that M. perniciosa able to utilize the methanol released from pectin demethylation; however, this metabolism is probably extracellular due to the presence of secreted MOX and CAT. Moreover, M. perniciosa possesses other cats, which are differentially expressed in vitro and in planta. Finally, we present a preliminary analysis of M. perniciosa secretome Doutorado Genética de Microorganismos Doutor em Genética e Biologia Molecular
- Published
- 2012
24. Genetic and metabolic study of Propionibacterium acidipropionici
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Maria Carolina de Barros Grassi, Pereira, Gonçalo Amarante Guimarães, 1964, Rincones, Johana, Padilha, Gabriel, Brocchi, Marcelo, Teixeira, Marcelo Menossi, Araújo, Welington Luiz de, Universidade Estadual de Campinas. Instituto de Biologia, Programa de Pós-Graduação em Genética e Biologia Molecular, and UNIVERSIDADE ESTADUAL DE CAMPINAS
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Genetic transformation ,Transformação genética ,Genome ,Propionibacterium ,1-Propanol ,Ácido propiônico ,Propionic acid ,Genomas - Abstract
Orientadores: Gonçalo Amarante Guimarães Pereira, Johana Rincones Perez Tese (doutorado) - Universidade Estadual de Campinas, Instituto de Biologia Resumo: O resumo poderá ser visualizado no texto completo da tese digital Abstract: The abstract is available with the full electronic document Doutorado Genética de Microorganismos Doutor em Genética e Biologia Molecular
- Published
- 2012
25. Carbon source metabolic interactions during the development of witche's broom disease of cocoa
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Joan Grande Barau, Pereira, Gonçalo Amarante Guimarães, 1964, Rincones, Johana, Queiroz, Marisa Vieira de, Rossi Filho, Antonio, Papes, Fabio, Teixeira, Marcelo Menossi, Universidade Estadual de Campinas. Instituto de Biologia, Programa de Pós-Graduação em Genética e Biologia Molecular, and UNIVERSIDADE ESTADUAL DE CAMPINAS
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Moniliophthora perniciosa ,Cocoa ,Xylem ,Glicerina ,Glycerin ,Xilema ,Cacau ,Vassoura-de-bruxa (Fitopatologia) ,Witche's broom disease (Phytopathology) - Abstract
Orientadores: Gonçalo Amarante Guimarães Pereira, Johana Rincones Peres Tese (doutorado) - Universidade Estadual de Campinas, Instituto de Biologia Resumo: A introdução da doença vassoura de bruxa do cacaueiro no Sul da Bahia representa um dos maiores exemplos da magnitude de impacto sócio econômico que uma fitopatologia pode aflingir a uma região ou país com um predominante perfil agroexportador. Com a execução do projeto genoma do fungo causador da vassoura de bruxa, o Basidiomiceto Moniliopthora perniciosa, uma série de trabalhos de caracterização morfológica, fisiológica e genética da doença foram inciados. Em um primeiro momento, em paralelo com a caracterização das alterações bioquímicas da doença, buscamos entender as diferenças de expressão gênica através de microarranjos de DNA e bibliotecas ESTs comparando as duas fases morfo-genéticas do fungo reproduzidas in vitro. Apesar de esse esforço pioneiro ter permitido a formação de um modelo panorâmico das alterações genéticas do fungo e fornecido importantes diretrizes para a pesquisa futura, ficou claro a necessidade de se explorar em maior detalhe a interação in vivo. Dentre os aspectos importantes investigados, o estudo do metabolismo do glicerol durante a fase biotrófica da interação através de marcadores de expressão mostrou que essa fonte de carbono em potencial pode ter um papel importante no início da infecção e participar da sinalização de stress através da modulação do shuttle mitocondrial de glicerol 3-fosfato. No entanto, a necessidade de se entender melhor o panorama das fontes de carbono disponíveis no apoplasto, o microambiente ocupado pelo fungo nas fases iniciais da interação, nos motivou a caracterizar as alterações nutricionais desse espaço ao longo da doença. Foi possível demonstrar que a vassoura de bruxa quebra o ciclo de carbono normal do apoplasto e que induz o corte da comunicação vascular com as partes sadias da planta ao final da fase biotrófica. Uma correlação entre a sinalização por carbono e a expressão de genes de patogenicidade secundária foi explorada. Com a expansão dos dados de genoma e transcriptoma utilizando sequenciamento de DNA de terceira geração (Illumina e 454 Life Sciences), dezenas de genes-alvo com potencial participação em vias de patogenicidade e desenvolvimento da doença foram identificados. Nesse contexto, dentre as ferramentas para análises de genômica funcional nesse modelo, a inativação ou nocaute de genes por gene targeting e o estudo da atividade de proteínas através de fusões com genes repórteres são essenciais ao teste das hipóteses geradas como fruto da pesquisa genômica da Vassoura-de-bruxa. Transformantes de M. perniciosa carregando uma cópia do gene marcador de autofagia MpATG8 fusionado ao marcador GFP apresentaram um padrão de fluorescência característico da biologia celular de ATG8, permitindo acompanhar o processo de autofagia sob condições de deprivação de nutrientes, demonstrando a possibilidade da aplicação de fusões com repórteres fluorescentes para experimentos de localização subcelular de proteínas desse fitopatógeno. Por fim, uma estratégia baseada na seleção de recombinantes através da técnica split marker em conjunto com um silenciamento transiente do sistema de reparo NHEJ por RNAi foi traçada e executada de modo a obter o nocaute do gene MpKu70, essencial a essa via de reparo, cuja inativação está associada a um aumento da eficiência de recombinação homóloga e gene targeting Abstract: Cocoa's Witches Broom Disease (WBD) is caused by the hemibiotrophic interaction between the Basidiomycete fungus Moniliopthora perniciosa and plants of Theobroma cacao. One remarkable feature of WBD is its unusual long biotrophic phase in which the fungal growth is restricted to the apoplastic space of plants. We compared differential gene expression between biotrophic and saprotrophic-like hyphae in vitro and proposed based on microarray and EST results that the maintenance of biotrophic-like cultures of M. perniciosa in vitro is subject to Carbon source regulation, especially regarding the use of glycerol as a preferential carbon source for biotrophy. To further gain insight into the carbon source regulation of WBD biotrophic phase in vivo, we characterized glycerol utilization using fungal expression markers during the initial phases of infection. We also characterized the carbon source fluctuations in the apoplast of healthy and infected cocoa seedlings. We showed that healthy cocoa plants possess a rhythmic carbon cycle in the apoplast that is in fine tune with periodical leaf flushing, expansion and photosynthetic maturation. WBD completely disrupts the carbon cycle in the apoplast of infected plants and also arrests leaf photosynthesis at pre sink-to-source transition levels. It was also possible to distinguish two different phases of carbon availability in the apoplast. The two phases correlated with fungal growth rates, the expression of a characterized necrosis inducing gene and the development of a tumor-like callus at the base of infected stems that completely disrupted normal vascular communication between healthy and diseased parts of the plant. A possible correlation between carbon starvation and phase switch from biotrophic to necrotrophic was explored and the evolution of the vascular-disrupting callus was discussed in the view of the tradeoff between the physiological behavior of infected tissues as sinks and the avoidance of nutrient remobilization during infected tissue senescence. Finally, we focused on the development of functional genomic tools for the study of gene function in this pathogen. As a proof-of-concept, we successfully obtained fungal strains expressing the autophagy related gene MpATG8 fused with GFP reporter, in which was possible to visualize the autophagy process in vivo by fluorescence imaging. In addition we also obtained the first successful gene targeting knockout of MpKu70, an M. perniciosa homologous of a gene essential to NHEJ in Eukaryotes. MpKu70 knockouts strains have reported phenotypes of increased homologous recombination, and the M. perniciosa strain may be an important platform for future functional studies of genes important in witches broom disease pathogenesis Doutorado Genética de Microorganismos Doutor em Genética e Biologia Molecular
- Published
- 2011
26. The mitochondrial alternative oxidase of Moniliophthora perniciosa and Moniliophthora roreri : its possible function in fungal development and pathogenesis
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Daniela Paula de Toledo Thomazella, Pereira, Gonçalo Amarante Guimarães, 1964, Rincones, Johana, Monteiro, Gisele, Soares Netto, Luis Eduardo, Teixeira, Marcelo Menossi, Arruda, Paulo, Universidade Estadual de Campinas. Instituto de Biologia, Programa de Pós-Graduação em Genética e Biologia Molecular, and UNIVERSIDADE ESTADUAL DE CAMPINAS
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Moniliophthora perniciosa ,Mitocôndria ,Moniliophthora roreri ,Mitochondria - Abstract
Orientadores: Gonçalo Amarante Guimarães Pereira, Johana Rincones Perez Tese (doutorado) - Universidade Estadual de Campinas, Instituto de Biologia Resumo: Moniliophihora perniciosa e Monüiophthora roreri são basidiomicetos fitopatogênicos. causadores de duas das mais devastadoras doenças fúngicas do cacaueiro: a Vassoura de bruxa e a Frosty Pod Rot (Momlíase), respectivamente. Apesar do grande impacto socioeconómico dessas doenças, aspectos importantes relacionados ao metabolismo destes patógenos ainda não são suficientemente compreendidos O estudo do metabolismo mitocondrial é particularmente relevante, pois compreende processos celulares vitais para a sobrevivência do organismo, como a regulação da produção de energia (ATP) e a manutenção do balanço redox da célula. Este trabalho teve como objetivo entender a importância da enzima mitocondrial oxidase alternativa (AOX) no metabolismo e desenvolvimento dos fungos M. perniciosa e M roreri- A AOX é uma oxidase terminal capaz de realizar o transporte de elétrons mitocondrial, catalisando a redução do oxigênio a água, de maneira desacoplada da síntese de ATP. Em M. perniciosa, a expressão do gene aox (Mp-aox) mostrou-se bastante relacionada ao ciclo de vida hemibiotrófico deste fungo. Níveis de expressão relativamente elevados deste gene foram observados no micélio biotrófico cultivado in vitro. De acordo, grande sensibilidade à inibição da via alternativa também foi verificada nesta fase micelial, sugerindo que a AOX tem participação no desenvolvimento deste primeiro estágio de M. perniciosa. Curiosamente, a inibição in vitro da cadeia respiratória principal retardou a transição da fase biotrófica para a necrotrófíca. Além disso, o uso combinado de inibidores da via principal e da AOX resultou em total inibição do desenvolvimento do fungo in vitro como também preveniu o estabelecimento da doença em plântulas de cacau. Com base nesses dados, o presente trabalho sugere um modelo no qual a participação diferencial das vias de transporte de elétrons (principal e alternativa) regula o desenvolvimento do micélio biotrófico e a transição de fase de M. perniciosa. Este é o primeiro trabalho em que se apresentam fortes evidências para uma função da AOX no desenvolvimento in planta e ciclo de vida de um fungo fítopatogênico. Como em M. perniciosa, o gene aox de M. roreri (Mr-aox) foi induzido em resposta ao fungicida azoxistrobina (inibidor do complexo III), sugerindo que a AOX tem participação na proteção do patògeno contra o estresse oxidativo gerado pela inibição da via principal- Apesar da indução do gene Mr-aox, M. roreri mostrou-se consideravelmente sensível ao fungicida. Deste modo, é possível que a ativação da AOX em M. roreri não seja suficiente para contornar os efeitos tóxicos da azoxistrobina e, consequentemente, que este fungo seja mais suscetível ao estresse oxidativo gerado pelo inibidor. Finalmente, a adição conjunta da azoxistrobina com um inibidor da AOX inibiu completamente o desenvolvimento in vitro de M. roreri. Com base nos resultados apresentados, sugere-se que a AOX tenha uma função relevante no metabolismo destes patógenos e que a inibição conjunta das vias principal e alternativa seja uma estratégia em potencial para o controle das doenças Vassoura de bruxa e Frosty PodRot. Abstract: Moniliophthora perniciosa and Moniliophthora roreri are phytopathogenic basidiomycetes, which cause two of the most devastating fungal diseases of cacao: the Witches' Broom and Frosty Pod Rot diseases, respectively. Despite the great socioeconomic impact of the diseases, important aspects of the metabolism of these pathogens are not sufficiently understood. The mitochondrial metabolism is particularly important since it is associated with many cellular processes of crucial importance for the organism survival, such as the regulation of energy production (ATP) and the maintenance of the cellular redox balance. This study aimed to understand the importance of the mitochondrial enzyme alternative oxidase (AOX) in the metabolism and development of the cacao pathogens M perniciosa and M. roreri. AOX is a non-phosporylating ubiquinol oxidase which catalyzes the reduction of molecular oxygen to water. In M. perniciosa, the expression of aox gene {Mp-aox) was closely related to the hemibiotrophic lifestyle of this fungus. High levels of Mp-aox transcripts were observed in the biotrophic mycelium and, accordingly, it showed an elevated sensitivity to AOX inhibitors, suggesting that AOX has a role in the development of the biotrophic phase. Interestingly, the in vitro inhibition of the cytochrome-dependent pathway prevented the transition from biotrophy to necrotrophy. Furthermore, the combined use of an inhibitor of the cytochrome pathway with an AOX inhibitor completely impaired the in vitro fungal growth as well as prevented the establishment of the disease in cacao seedlings. Based on these data, this study suggests a model in which the involvement of the different pathways of electron transfer (cytochrome and alternative routes) regulates the development of the biotrophic mycelium and the phase transition of M. perniciosa in planta. This is the first report presenting strong evidences for a role of AOX during the in pfanta development and life cycle of a phytopathogenic fijngus. Like M. perniciosa, the aox gene of M. roreri {Mr-aox) is up-regulated after exposure of the fungal mycelium to the fungicide azoxystrobin (a potent inhibitor of complex III}, suggesting that AOX plays a protective role against the deleterious oxidative stress produced by the compound. However, even with the increased expression of Mr-aox, M. roreri growth was highly sensitive to the fungicide. Therefore, it is possible that AOX activation was not enough to overcome the toxic effects of the cytochrome pathway inhibition and, consequently, M. roreri presented a high susceptibility to oxidative stress. Finally, the combined application of azoxystrobin and an AOX inhibitor completely impaired the in vitro development of M. roreri Thus., based on these results, we suggest that AOX has a role in the metabolism of these cacao pathogens and the concomitant inhibition of both cytochrome and alternative pathways may be an efficient strategy for the control of Witches' Broom and Frosty Pod Rot diseases. Doutorado Genética de Microorganismos Doutor em Genética e Biologia Molecular
- Published
- 2010
27. The glyceraldehyde-3-phosphate dehydrogenase gene of Moniliophthoraperniciosa, the causal agent of witches' broom disease of Theobroma cacao.
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Lima JO, Pereira JF, Rincones J, Barau JG, Araújo EF, Pereira GA, and Queiroz MV
- Abstract
This report describes the cloning, sequence and expression analysis of the glyceraldehyde-3-phosphate dehydrogenase (GAPDH) gene of Moniliophthora perniciosa, the most important pathogen of cocoa in Brazil. Southern blot analysis revealed the presence of a single copy of the GAPDH gene in the M. perniciosa genome (MpGAPDH). The complete MpGAPDH coding sequence contained 1,461 bp with eight introns that were conserved in the GAPDH genes of other basidiomycete species. The cis-elements in the promoter region of the MpGAPDH gene were similar to those of other basidiomycetes. Likewise, the MpGAPDH gene encoded a putative 339 amino acid protein that shared significant sequence similarity with other GAPDH proteins in fungi, plants, and metazoans. Phylogenetic analyses clustered the MPGAPDH protein with other homobasidiomycete fungi of the family Tricholomataceae. Expression analysis of the MpGAPDH gene by real-time PCR showed that this gene was more expressed (~1.3X) in the saprotrophic stage of this hemibiotrophic plant pathogen than in the biotrophic stage when grown in cacao extracts.
- Published
- 2009
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- View/download PDF
28. The mitochondrial genome of the phytopathogenic basidiomycete Moniliophthora perniciosa is 109 kb in size and contains a stable integrated plasmid.
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Formighieri EF, Tiburcio RA, Armas ED, Medrano FJ, Shimo H, Carels N, Góes-Neto A, Cotomacci C, Carazzolle MF, Sardinha-Pinto N, Thomazella DP, Rincones J, Digiampietri L, Carraro DM, Azeredo-Espin AM, Reis SF, Deckmann AC, Gramacho K, Gonçalves MS, Moura Neto JP, Barbosa LV, Meinhardt LW, Cascardo JC, and Pereira GA
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- Agaricales classification, Amino Acid Sequence, Base Composition, Base Sequence, Chromosome Mapping, Codon, Introns, Mitochondrial Proteins chemistry, Mitochondrial Proteins genetics, Molecular Sequence Data, Open Reading Frames, Phylogeny, Agaricales chemistry, Agaricales genetics, Cacao microbiology, Genome, Mitochondrial, Plant Diseases microbiology, Plasmids genetics
- Abstract
We present here the sequence of the mitochondrial genome of the basidiomycete phytopathogenic hemibiotrophic fungus Moniliophthora perniciosa, causal agent of the Witches' Broom Disease in Theobroma cacao. The DNA is a circular molecule of 109,103 base pairs, with 31.9% GC, and is the largest sequenced so far. This size is due essentially to the presence of numerous non-conserved hypothetical ORFs. It contains the 14 genes coding for proteins involved in the oxidative phosphorylation, the two rRNA genes, one ORF coding for a ribosomal protein (rps3), and a set of 26 tRNA genes that recognize codons for all amino acids. Seven homing endonucleases are located inside introns. Except atp8, all conserved known genes are in the same orientation. Phylogenetic analysis based on the cox genes agrees with the commonly accepted fungal taxonomy. An uncommon feature of this mitochondrial genome is the presence of a region that contains a set of four, relatively small, nested, inverted repeats enclosing two genes coding for polymerases with an invertron-type structure and three conserved hypothetical genes interpreted as the stable integration of a mitochondrial linear plasmid. The integration of this plasmid seems to be a recent evolutionary event that could have implications in fungal biology. This sequence is available under GenBank accession number AY376688.
- Published
- 2008
- Full Text
- View/download PDF
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