Caracterização molecular da resistência à ferrugem asiática da soja mediada pelo gene Rpp4

Detalhes bibliográficos
Ano de defesa: 2011
Autor(a) principal: Morales, Aguida Maria Alves Pereira
Orientador(a): Oliveira, Aluízio Borém de lattes
Banca de defesa: Loureiro, Marcelo Ehlers lattes, Paula Júnior, Trazilbo José de lattes, Brommonschenkel, Sérgio Hermínio lattes
Tipo de documento: Tese
Tipo de acesso: Acesso aberto
Idioma: por
Instituição de defesa: Universidade Federal de Viçosa
Programa de Pós-Graduação: Doutorado em Genética e Melhoramento
Departamento: Genética animal; Genética molecular e de microrganismos; Genética quantitativa; Genética vegetal; Me
País: BR
Palavras-chave em Português:
Palavras-chave em Inglês:
Área do conhecimento CNPq:
Link de acesso: http://locus.ufv.br/handle/123456789/1405
Resumo: The main control method used is the application of fungicides, which substantially increase the cost of production and are harmful to the environment. Prevention is still the best way to avoid more significant losses in soybean production. Alternatives, such as planting resistant varieties to the fungus, are also important. The use of resistant or tolerant varieties is the most promising method for control of Asian soybean rust. Recently, five single dominant genes to specific soybean rust isolates were described: Rpp1, Rpp2, Rpp3, Rpp4 and Rpp5. However, little is known about the molecular interaction between soybean and soybean rust and on the molecular pathway triggered by pathogen recognition. Understanding the molecular mechanisms involved in defense responses is of primary importance in planning strategies for controlling stress and consequently increasing plant adaptation to limiting conditions. Previously, Rpp4 was mapped to soybean linkage group G (chromosome 18), and the sequencing of this region in the susceptible genotype Williams 82 (Wm82) and resistant (Pl459025B) identified a cluster of CC- NBS-LRR resistance genes. Meyers et al. (2009) developed Virus lnduced Gene Silencing constructs from the NBD and LRR regions of the Wm82 Rpp4 Candidate genes to test whether paralogous genes were responsible for resistance in the resistant parent (Pl459025B). ln this study, RNA extracted from the same Rpp4 LRR silenced and empty vector treated plants described by Meyer et al. (2009) was compared. Transcriptome analyses of three independent biological replicates was performed using the GeneChip® Soybean Genome Array (Affymetrix®). A total of 383 genes were found to be significantly differentially expressed between Rpp4 silenced and no silenced plants infected with P. pachyrhizi. Of the 383 differentially expressed genes, 22 were induced, and 361 were suppressed. Using Clover (cis-element over representation) software and the TRANSFAC (transcription factor database we identified 33 transcription factor-binding sites significantly over represented in our differentially expressed genes when compared to all genes in the soybean genome, several with known roles in defense. Finally, to elucidate which genes are exclusively related to Rpp4-mediated signaling, we have compared the results of our experiment with microarray results from Rpp2, Rpp3 and Rpp4 resistant and susceptible reactions. We have identified 101 genes unique to the Rpp4-signaling pathway. ln addition, in order to obtain more information about Rpp4 function, we used real time quantitative PCR (RT-qPCR) to analyze the expression of all Rpp4 genes in different plant tissues, in different stages of development and after inoculation with P. pachyrhizi. We have developed a single pair of primers from the NBD domain that allow us to monitor the expression of all ten genes. Direct sequencing of the RT-qPCR product differentiates between the ten genes. ln addition we examined the occurrence of alternative splice Rpp4 gene under inoculation effect.
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spelling Morales, Aguida Maria Alves Pereirahttp://lattes.cnpq.br/8213472808311219Abdelnoor, Ricardo Vilelahttp://lattes.cnpq.br/7381930992034213Sakiyama, Ney Sussumuhttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4781483H8Oliveira, Aluízio Borém dehttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4783555Z3Loureiro, Marcelo Ehlershttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4780851Y3Paula Júnior, Trazilbo José dehttp://lattes.cnpq.br/7899276097018876Brommonschenkel, Sérgio Hermíniohttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4780948Y42015-03-26T12:45:44Z2014-10-102015-03-26T12:45:44Z2011-07-18MORALES, Aguida Maria Alves Pereira. Molecular characterization of resistance to Asian soybean rust mediated by Rpp4. 2011. 84 f. Tese (Doutorado em Genética animal; Genética molecular e de microrganismos; Genética quantitativa; Genética vegetal; Me) - Universidade Federal de Viçosa, Viçosa, 2011.http://locus.ufv.br/handle/123456789/1405The main control method used is the application of fungicides, which substantially increase the cost of production and are harmful to the environment. Prevention is still the best way to avoid more significant losses in soybean production. Alternatives, such as planting resistant varieties to the fungus, are also important. The use of resistant or tolerant varieties is the most promising method for control of Asian soybean rust. Recently, five single dominant genes to specific soybean rust isolates were described: Rpp1, Rpp2, Rpp3, Rpp4 and Rpp5. However, little is known about the molecular interaction between soybean and soybean rust and on the molecular pathway triggered by pathogen recognition. Understanding the molecular mechanisms involved in defense responses is of primary importance in planning strategies for controlling stress and consequently increasing plant adaptation to limiting conditions. Previously, Rpp4 was mapped to soybean linkage group G (chromosome 18), and the sequencing of this region in the susceptible genotype Williams 82 (Wm82) and resistant (Pl459025B) identified a cluster of CC- NBS-LRR resistance genes. Meyers et al. (2009) developed Virus lnduced Gene Silencing constructs from the NBD and LRR regions of the Wm82 Rpp4 Candidate genes to test whether paralogous genes were responsible for resistance in the resistant parent (Pl459025B). ln this study, RNA extracted from the same Rpp4 LRR silenced and empty vector treated plants described by Meyer et al. (2009) was compared. Transcriptome analyses of three independent biological replicates was performed using the GeneChip® Soybean Genome Array (Affymetrix®). A total of 383 genes were found to be significantly differentially expressed between Rpp4 silenced and no silenced plants infected with P. pachyrhizi. Of the 383 differentially expressed genes, 22 were induced, and 361 were suppressed. Using Clover (cis-element over representation) software and the TRANSFAC (transcription factor database we identified 33 transcription factor-binding sites significantly over represented in our differentially expressed genes when compared to all genes in the soybean genome, several with known roles in defense. Finally, to elucidate which genes are exclusively related to Rpp4-mediated signaling, we have compared the results of our experiment with microarray results from Rpp2, Rpp3 and Rpp4 resistant and susceptible reactions. We have identified 101 genes unique to the Rpp4-signaling pathway. ln addition, in order to obtain more information about Rpp4 function, we used real time quantitative PCR (RT-qPCR) to analyze the expression of all Rpp4 genes in different plant tissues, in different stages of development and after inoculation with P. pachyrhizi. We have developed a single pair of primers from the NBD domain that allow us to monitor the expression of all ten genes. Direct sequencing of the RT-qPCR product differentiates between the ten genes. ln addition we examined the occurrence of alternative splice Rpp4 gene under inoculation effect.Práticas de gerenciamento são essenciais para o controle da ferrugem. O principal método de controle utilizado é a aplicação de fungicida, o qual aumenta substancialmente o custo de produção e são prejudiciais ao ambiente. A prevenção ainda é a melhor maneira de evitar perdas na produção de soja. Alternativas como plantar cultivares resistentes ao fungo também são importantes. O uso de variedades resistentes ou tolerantes é o método mais promissor para o controle da ferrugem asiática, cinco locus dominantes de resistência tem sido descrito na literatura: Rpp 1, Rpp2, Rpp3, Rpp4 e Rpp5. Entretanto pouco se sabe sobre a interação molecular desencadeada pela reconhecimento do patógeno pela planta em relação a ferrugem asiática. Entender os mecanismos moleculares envolvidos na resposta de defesa é de primária importância no planejamento de estratégias de controle do estresse e para consequente aumento da adaptação da planta. O Rpp4 foi mapeado no grupo de ligação G da soja (cromossomo 18) e o sequenciamento desta região no genótipo suscetível Williams82 (Wm82) e resistente (Pl459025B) identificou um cluster de genes de resistência CC-NBS- LRR. Meyers et al., (2009) desenvolveu construções para silenciamento induzido por vírus a partir das regiões NBD e LRR dos genes candidatos Rpp4 no genótipo Wm82 para testar se os genes parãlogos são responsáveis pela resistência no genótipo resistente (Pl459025B). Neste estudo, o RNA foi extraído de plantas silenciadas Rpp4LRR e de plantas controle (Vetor vazio). Análises transcricional de 3 replicatas biológicas foi feita utilizando GeneChip® Soybean Genome Array (Affymetrix®). Um total de 383 genes foram encontrados ser diferencialmente expressos entre plantas Rpp4 silenciadas e plantas não silenciadas (controle) quando infectadas com P. pachyrhizi. Dos 383 genes diferencialmente expressos, 22 foram induzidos e 361 foram reprimidos. Além disso, utilizando a ferramenta Clover (cis-element over representation) e TRANSFAC (transcription factor database) identificamos 33 sitios para fatores de transcrição presentes nos promotores dos genes diferencialmente expressos. Finalmente, para elucidar quais os genes são exclusivamentes mediados pela sinalização do Rpp4, nós comparamos os resultados de nosso experimento com os resultados de microarranjos oriundos de Rpp2, Rpp3 e Rpp4 resistente e suscetível. Nós identificamos 101 genes exclusivos. Além disso, com o objetivo de se obter maior informação sobre a função do Rpp4 nós utilizamos PCR quantitativo em tempo real (RT-qPCR) para analisar a expressão de todos os genes Rpp4 em diferentes tecidos da planta, diferentes estágios de desenvolvimento e depois da inoculação com P. pachyrhizi. Nós desenvolvemos um par de primers no domínio NBD que nós permitiu monitorar a expressão de todos os genes. O sequenciamento direto dos produtos originados no RT-qPCR nos permitiu diferenciar entre os 10 genes. Além disso nós examinamos a ocorrência de splice alternativo do gene Rpp4 na soja sob efeito da inoculação.Conselho Nacional de Desenvolvimento Científico e Tecnológicoapplication/pdfporUniversidade Federal de ViçosaDoutorado em Genética e MelhoramentoUFVBRGenética animal; Genética molecular e de microrganismos; Genética quantitativa; Genética vegetal; MeSojaFerrugem-da-sojaResistência a doenças e pragasBiologia molecularSoybeansOf soybean rustResistance to diseases and pestsMolecular biologyCNPQ::CIENCIAS BIOLOGICAS::GENETICA::GENETICA VEGETALCaracterização molecular da resistência à ferrugem asiática da soja mediada pelo gene Rpp4Molecular characterization of resistance to Asian soybean rust mediated by Rpp4info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesisinfo:eu-repo/semantics/openAccessreponame:LOCUS Repositório Institucional da UFVinstname:Universidade Federal de Viçosa (UFV)instacron:UFVORIGINALtexto completo.pdfapplication/pdf1512138https://locus.ufv.br//bitstream/123456789/1405/1/texto%20completo.pdfd84e1b8e0731f063bf7efac1b010836cMD51TEXTtexto completo.pdf.txttexto completo.pdf.txtExtracted texttext/plain153154https://locus.ufv.br//bitstream/123456789/1405/2/texto%20completo.pdf.txtce550101505bb07f491a7759a5a3a444MD52THUMBNAILtexto completo.pdf.jpgtexto completo.pdf.jpgIM Thumbnailimage/jpeg3783https://locus.ufv.br//bitstream/123456789/1405/3/texto%20completo.pdf.jpg6cb18f7f60996d28b394bde880010e60MD53123456789/14052016-04-07 23:07:33.124oai:locus.ufv.br:123456789/1405Repositório InstitucionalPUBhttps://www.locus.ufv.br/oai/requestfabiojreis@ufv.bropendoar:21452016-04-08T02:07:33LOCUS Repositório Institucional da UFV - Universidade Federal de Viçosa (UFV)false
dc.title.por.fl_str_mv Caracterização molecular da resistência à ferrugem asiática da soja mediada pelo gene Rpp4
dc.title.alternative.eng.fl_str_mv Molecular characterization of resistance to Asian soybean rust mediated by Rpp4
title Caracterização molecular da resistência à ferrugem asiática da soja mediada pelo gene Rpp4
spellingShingle Caracterização molecular da resistência à ferrugem asiática da soja mediada pelo gene Rpp4
Morales, Aguida Maria Alves Pereira
Soja
Ferrugem-da-soja
Resistência a doenças e pragas
Biologia molecular
Soybeans
Of soybean rust
Resistance to diseases and pests
Molecular biology
CNPQ::CIENCIAS BIOLOGICAS::GENETICA::GENETICA VEGETAL
title_short Caracterização molecular da resistência à ferrugem asiática da soja mediada pelo gene Rpp4
title_full Caracterização molecular da resistência à ferrugem asiática da soja mediada pelo gene Rpp4
title_fullStr Caracterização molecular da resistência à ferrugem asiática da soja mediada pelo gene Rpp4
title_full_unstemmed Caracterização molecular da resistência à ferrugem asiática da soja mediada pelo gene Rpp4
title_sort Caracterização molecular da resistência à ferrugem asiática da soja mediada pelo gene Rpp4
author Morales, Aguida Maria Alves Pereira
author_facet Morales, Aguida Maria Alves Pereira
author_role author
dc.contributor.authorLattes.por.fl_str_mv http://lattes.cnpq.br/8213472808311219
dc.contributor.author.fl_str_mv Morales, Aguida Maria Alves Pereira
dc.contributor.advisor-co1.fl_str_mv Abdelnoor, Ricardo Vilela
dc.contributor.advisor-co1Lattes.fl_str_mv http://lattes.cnpq.br/7381930992034213
dc.contributor.advisor-co2.fl_str_mv Sakiyama, Ney Sussumu
dc.contributor.advisor-co2Lattes.fl_str_mv http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4781483H8
dc.contributor.advisor1.fl_str_mv Oliveira, Aluízio Borém de
dc.contributor.advisor1Lattes.fl_str_mv http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4783555Z3
dc.contributor.referee1.fl_str_mv Loureiro, Marcelo Ehlers
dc.contributor.referee1Lattes.fl_str_mv http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4780851Y3
dc.contributor.referee2.fl_str_mv Paula Júnior, Trazilbo José de
dc.contributor.referee2Lattes.fl_str_mv http://lattes.cnpq.br/7899276097018876
dc.contributor.referee3.fl_str_mv Brommonschenkel, Sérgio Hermínio
dc.contributor.referee3Lattes.fl_str_mv http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4780948Y4
contributor_str_mv Abdelnoor, Ricardo Vilela
Sakiyama, Ney Sussumu
Oliveira, Aluízio Borém de
Loureiro, Marcelo Ehlers
Paula Júnior, Trazilbo José de
Brommonschenkel, Sérgio Hermínio
dc.subject.por.fl_str_mv Soja
Ferrugem-da-soja
Resistência a doenças e pragas
Biologia molecular
topic Soja
Ferrugem-da-soja
Resistência a doenças e pragas
Biologia molecular
Soybeans
Of soybean rust
Resistance to diseases and pests
Molecular biology
CNPQ::CIENCIAS BIOLOGICAS::GENETICA::GENETICA VEGETAL
dc.subject.eng.fl_str_mv Soybeans
Of soybean rust
Resistance to diseases and pests
Molecular biology
dc.subject.cnpq.fl_str_mv CNPQ::CIENCIAS BIOLOGICAS::GENETICA::GENETICA VEGETAL
description The main control method used is the application of fungicides, which substantially increase the cost of production and are harmful to the environment. Prevention is still the best way to avoid more significant losses in soybean production. Alternatives, such as planting resistant varieties to the fungus, are also important. The use of resistant or tolerant varieties is the most promising method for control of Asian soybean rust. Recently, five single dominant genes to specific soybean rust isolates were described: Rpp1, Rpp2, Rpp3, Rpp4 and Rpp5. However, little is known about the molecular interaction between soybean and soybean rust and on the molecular pathway triggered by pathogen recognition. Understanding the molecular mechanisms involved in defense responses is of primary importance in planning strategies for controlling stress and consequently increasing plant adaptation to limiting conditions. Previously, Rpp4 was mapped to soybean linkage group G (chromosome 18), and the sequencing of this region in the susceptible genotype Williams 82 (Wm82) and resistant (Pl459025B) identified a cluster of CC- NBS-LRR resistance genes. Meyers et al. (2009) developed Virus lnduced Gene Silencing constructs from the NBD and LRR regions of the Wm82 Rpp4 Candidate genes to test whether paralogous genes were responsible for resistance in the resistant parent (Pl459025B). ln this study, RNA extracted from the same Rpp4 LRR silenced and empty vector treated plants described by Meyer et al. (2009) was compared. Transcriptome analyses of three independent biological replicates was performed using the GeneChip® Soybean Genome Array (Affymetrix®). A total of 383 genes were found to be significantly differentially expressed between Rpp4 silenced and no silenced plants infected with P. pachyrhizi. Of the 383 differentially expressed genes, 22 were induced, and 361 were suppressed. Using Clover (cis-element over representation) software and the TRANSFAC (transcription factor database we identified 33 transcription factor-binding sites significantly over represented in our differentially expressed genes when compared to all genes in the soybean genome, several with known roles in defense. Finally, to elucidate which genes are exclusively related to Rpp4-mediated signaling, we have compared the results of our experiment with microarray results from Rpp2, Rpp3 and Rpp4 resistant and susceptible reactions. We have identified 101 genes unique to the Rpp4-signaling pathway. ln addition, in order to obtain more information about Rpp4 function, we used real time quantitative PCR (RT-qPCR) to analyze the expression of all Rpp4 genes in different plant tissues, in different stages of development and after inoculation with P. pachyrhizi. We have developed a single pair of primers from the NBD domain that allow us to monitor the expression of all ten genes. Direct sequencing of the RT-qPCR product differentiates between the ten genes. ln addition we examined the occurrence of alternative splice Rpp4 gene under inoculation effect.
publishDate 2011
dc.date.issued.fl_str_mv 2011-07-18
dc.date.available.fl_str_mv 2014-10-10
2015-03-26T12:45:44Z
dc.date.accessioned.fl_str_mv 2015-03-26T12:45:44Z
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dc.identifier.citation.fl_str_mv MORALES, Aguida Maria Alves Pereira. Molecular characterization of resistance to Asian soybean rust mediated by Rpp4. 2011. 84 f. Tese (Doutorado em Genética animal; Genética molecular e de microrganismos; Genética quantitativa; Genética vegetal; Me) - Universidade Federal de Viçosa, Viçosa, 2011.
dc.identifier.uri.fl_str_mv http://locus.ufv.br/handle/123456789/1405
identifier_str_mv MORALES, Aguida Maria Alves Pereira. Molecular characterization of resistance to Asian soybean rust mediated by Rpp4. 2011. 84 f. Tese (Doutorado em Genética animal; Genética molecular e de microrganismos; Genética quantitativa; Genética vegetal; Me) - Universidade Federal de Viçosa, Viçosa, 2011.
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dc.publisher.country.fl_str_mv BR
dc.publisher.department.fl_str_mv Genética animal; Genética molecular e de microrganismos; Genética quantitativa; Genética vegetal; Me
publisher.none.fl_str_mv Universidade Federal de Viçosa
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