Caracterização molecular da resistência à ferrugem asiática da soja mediada pelo gene Rpp4
Ano de defesa: | 2011 |
---|---|
Autor(a) principal: | |
Orientador(a): | |
Banca de defesa: | , , |
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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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 |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/doctoralThesis |
format |
doctoralThesis |
status_str |
publishedVersion |
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. |
url |
http://locus.ufv.br/handle/123456789/1405 |
dc.language.iso.fl_str_mv |
por |
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por |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
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openAccess |
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Universidade Federal de Viçosa |
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Doutorado em Genética e Melhoramento |
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UFV |
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 |
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Universidade Federal de Viçosa |
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