Caracterização molecular e inoculação de Ralstonia solanacearum em Eucalyptus spp
Autor(a) principal: | |
---|---|
Data de Publicação: | 2012 |
Tipo de documento: | Dissertação |
Idioma: | por |
Título da fonte: | LOCUS Repositório Institucional da UFV |
Texto Completo: | http://locus.ufv.br/handle/123456789/4412 |
Resumo: | The Ralstonia wilt of eucalyptus, caused by Ralstonia solanacearum, is potentially one of the major disease of culture. Because of its high genetic and phenotypic variability, currently R. solanacearum is considered a "species complex", subdivided into four taxonomic levels: species, filotipo, sequevar and clone. In order to understand the variability among isolates of R. solanacearum from eucalyptus and develop a protocol for inoculation and screening of resistant clones of eucalyptus to Ralstonia wilt this work was performed. The work was divided into two articles: (i) Molecular characterization of isolates of Ralstonia solanacearum in Eucalyptus spp. and (ii) Method of inoculation and evaluation of resistant clones of Eucalyptus spp. to Ralstonia wilt caused by Ralstonia solanacearum. In Article 1, 19 isolates were analyzed from different regions of eucalyptus in Brazil based on new classification (filotipo and sequevar) and its genetic variability. A product of 372 bp generated by multiplex-PCR amplification using primers Nmult permitted the identification of all isolates analyzed as pertaining to filotipo II. Eighteen isolates were grouped in subclade IIA and just one isolate in IIB. The phylogenetic tree generated from the endoglucanase (egl) gene sequences confirmed the classification of the isolates in filotipo II and separated them into sequevares. The strains AMC22, IBSBF2568 and IBSBF2576 were grouped into a single clade, as well as isolates UFV18 and UFV20, with 89% and 78% posterior probability, respectively, forming two new possible sequevares not defined yet. We also identified isolates belonging to sequevar 41 (100% probability) and 37 (88% probability). However, most of the isolates did not fit in any of previously described sequevar, or formed defined clades. The analysis of results of obtained fragments amplified by ERIC-PCR technique suggested a great genetic diversity among the isolates analyzed in this work. In addition, a high correlation between the geographical origin of isolates and the similarity showed by them was observed. In Article 2, three methods of inoculation and resistant of Eucalyptus spp. clones to Ralstonia wilt was evaluated: (i) soil infestation with R. solanacearum, (ii) dipping of sectioned roots in bacterial suspension, and (iii) injection of a bacterial suspension in the base of the stem. The injection method of a bacterial suspension at the stem base was the most efficient inoculation method of R. solanacearum on eucalyptus. Four, out of 21 clones tested for Ralstonia wilt resistance by this method, were classified as resistant by not showing symptoms of wilt and bacterial exudation up to 30 days after inoculation. |
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Fonseca, Natália Rissohttp://lattes.cnpq.br/2709153501578306Lopes, Carlos Albertohttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4783372D6Cunha, Luis Claudio Vieira dahttp://lattes.cnpq.br/9367947173322278Alfenas, Acelino Coutohttp://lattes.cnpq.br/2514320654462590Guimarães, Lúcio Mauro da Silvahttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4766939H12015-03-26T13:37:49Z2013-04-232015-03-26T13:37:49Z2012-07-18FONSECA, Natália Risso. Molecular characterization and inoculation of Ralstonia solanacearum in Eucalyptus spp. 2012. 54 f. Dissertação (Mestrado em Etiologia; Epidemiologia; Controle) - Universidade Federal de Viçosa, Viçosa, 2012.http://locus.ufv.br/handle/123456789/4412The Ralstonia wilt of eucalyptus, caused by Ralstonia solanacearum, is potentially one of the major disease of culture. Because of its high genetic and phenotypic variability, currently R. solanacearum is considered a "species complex", subdivided into four taxonomic levels: species, filotipo, sequevar and clone. In order to understand the variability among isolates of R. solanacearum from eucalyptus and develop a protocol for inoculation and screening of resistant clones of eucalyptus to Ralstonia wilt this work was performed. The work was divided into two articles: (i) Molecular characterization of isolates of Ralstonia solanacearum in Eucalyptus spp. and (ii) Method of inoculation and evaluation of resistant clones of Eucalyptus spp. to Ralstonia wilt caused by Ralstonia solanacearum. In Article 1, 19 isolates were analyzed from different regions of eucalyptus in Brazil based on new classification (filotipo and sequevar) and its genetic variability. A product of 372 bp generated by multiplex-PCR amplification using primers Nmult permitted the identification of all isolates analyzed as pertaining to filotipo II. Eighteen isolates were grouped in subclade IIA and just one isolate in IIB. The phylogenetic tree generated from the endoglucanase (egl) gene sequences confirmed the classification of the isolates in filotipo II and separated them into sequevares. The strains AMC22, IBSBF2568 and IBSBF2576 were grouped into a single clade, as well as isolates UFV18 and UFV20, with 89% and 78% posterior probability, respectively, forming two new possible sequevares not defined yet. We also identified isolates belonging to sequevar 41 (100% probability) and 37 (88% probability). However, most of the isolates did not fit in any of previously described sequevar, or formed defined clades. The analysis of results of obtained fragments amplified by ERIC-PCR technique suggested a great genetic diversity among the isolates analyzed in this work. In addition, a high correlation between the geographical origin of isolates and the similarity showed by them was observed. In Article 2, three methods of inoculation and resistant of Eucalyptus spp. clones to Ralstonia wilt was evaluated: (i) soil infestation with R. solanacearum, (ii) dipping of sectioned roots in bacterial suspension, and (iii) injection of a bacterial suspension in the base of the stem. The injection method of a bacterial suspension at the stem base was the most efficient inoculation method of R. solanacearum on eucalyptus. Four, out of 21 clones tested for Ralstonia wilt resistance by this method, were classified as resistant by not showing symptoms of wilt and bacterial exudation up to 30 days after inoculation.A murcha-de-Ralstonia do eucalipto, causada por Ralstonia solanacearum, constitui potencialmente, uma das principais doenças da cultura. Devido à sua ampla variabilidade, atualmente R. solanacearum é considerada um complexo de espécies , subdividida em quatro níveis taxonômicos: espécie, filotipo, sequevar e clone. Com o intuito de entender a variabilidade entre isolados de R. solanacearum provenientes de eucalipto e desenvolver um protocolo de inoculação e avaliação da resistência de clones de eucalipto à murcha-de-Ralstonia realizou-se este trabalho. Ele foi dividido em dois artigos: (i) Caracterização molecular de isolados de Ralstonia solanacearum de Eucalyptus spp. e (ii) Método de inoculação e avaliação de resistência de clones de Eucalyptus spp. à murcha-de-Ralstonia causada por Ralstonia solanacearum. No artigo 1, foram analisados 19 isolados de R. solanacearum obtidos de eucalipto de diferentes regiões do Brasil quanto à nova classificação (filotipo e sequevar) e quanto à variabilidade genética. Um produto de 372 pb gerado pela amplificação por PCR-multiplex, utilizando primers Nmult, permitiu identificar todos os isolados analisados no filotipo II. Dezoito isolados foram agrupados no subclado IIA e apenas um no IIB. A árvore filogenética gerada a partir das sequências do gene da endoglucanase (egl) confirmou a classificação dos isolados no filotipo II e os separou em sequevares. Os isolados AMC22, IBSBF2568 e IBSBF2576 agruparam-se em um único clado, assim como os isolados UFV18 e UFV20, com 89% e 78% de probabilidade posteriori, respectivamente, compondo dois novos possíveis sequevares, ainda não definidos. Foram identificados também isolados pertencentes ao sequevar 41 (100% de probabilidade) e 37 (88% de probabilidade). Entretanto, a maioria dos isolados não se enquadrou em nenhum sequevar descrito, nem formaram clados definidos. O resultado da análise de fragmentos amplificados pela técnica de ERIC-PCR indicou grande diversidade genética entre os isolados avaliados neste trabalho, havendo, em geral, uma alta correlação entre a origem geográfica dos isolados e a similaridade entre eles. No artigo 2, avaliaram-se três métodos de inoculação e a resistência de clones de Eucalyptus spp. à murcha-de-Ralstonia: (i) infestação do solo com R. solanacearum; (ii) imersão de raízes seccionadas em suspensão bacteriana; e (iii) injeção de suspensão bacteriana na base do caule. O método de inoculação de injeção de suspensão bacteriana na base do caule destacou-se como o mais eficiente para inoculações de R. solanacearum em eucalipto. De 21 clones avaliados quanto à resistência à murcha-de-Ralstonia por esse método, apenas quatro foram classificados como resistentes por não apresentarem sintomas de murcha e exsudação bacteriana até 30 dias após a inoculação.Conselho Nacional de Desenvolvimento Científico e Tecnológicoapplication/pdfporUniversidade Federal de ViçosaMestrado em FitopatologiaUFVBREtiologia; Epidemiologia; ControleMurcha bacterianaRalstonia solanacearumEucaliptoResistênciaBacterial wiltRalstonia solanacearumEucalyptusResistanceCNPQ::CIENCIAS AGRARIAS::AGRONOMIA::FITOSSANIDADE::FITOPATOLOGIACaracterização molecular e inoculação de Ralstonia solanacearum em Eucalyptus sppMolecular characterization and inoculation of Ralstonia solanacearum in Eucalyptus sppinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisinfo:eu-repo/semantics/openAccessreponame:LOCUS Repositório Institucional da UFVinstname:Universidade Federal de Viçosa (UFV)instacron:UFVORIGINALtexto completo.pdfapplication/pdf1080649https://locus.ufv.br//bitstream/123456789/4412/1/texto%20completo.pdf0b734c22ca6fdd3994db4802005b6004MD51TEXTtexto completo.pdf.txttexto completo.pdf.txtExtracted texttext/plain96175https://locus.ufv.br//bitstream/123456789/4412/2/texto%20completo.pdf.txtd30295a61fbbb2edc10a9238cb7b2da5MD52THUMBNAILtexto completo.pdf.jpgtexto completo.pdf.jpgIM Thumbnailimage/jpeg3598https://locus.ufv.br//bitstream/123456789/4412/3/texto%20completo.pdf.jpg26a873669250614de063049fbf0f6817MD53123456789/44122016-04-10 23:09:46.396oai:locus.ufv.br:123456789/4412Repositório InstitucionalPUBhttps://www.locus.ufv.br/oai/requestfabiojreis@ufv.bropendoar:21452016-04-11T02:09:46LOCUS Repositório Institucional da UFV - Universidade Federal de Viçosa (UFV)false |
dc.title.por.fl_str_mv |
Caracterização molecular e inoculação de Ralstonia solanacearum em Eucalyptus spp |
dc.title.alternative.eng.fl_str_mv |
Molecular characterization and inoculation of Ralstonia solanacearum in Eucalyptus spp |
title |
Caracterização molecular e inoculação de Ralstonia solanacearum em Eucalyptus spp |
spellingShingle |
Caracterização molecular e inoculação de Ralstonia solanacearum em Eucalyptus spp Fonseca, Natália Risso Murcha bacteriana Ralstonia solanacearum Eucalipto Resistência Bacterial wilt Ralstonia solanacearum Eucalyptus Resistance CNPQ::CIENCIAS AGRARIAS::AGRONOMIA::FITOSSANIDADE::FITOPATOLOGIA |
title_short |
Caracterização molecular e inoculação de Ralstonia solanacearum em Eucalyptus spp |
title_full |
Caracterização molecular e inoculação de Ralstonia solanacearum em Eucalyptus spp |
title_fullStr |
Caracterização molecular e inoculação de Ralstonia solanacearum em Eucalyptus spp |
title_full_unstemmed |
Caracterização molecular e inoculação de Ralstonia solanacearum em Eucalyptus spp |
title_sort |
Caracterização molecular e inoculação de Ralstonia solanacearum em Eucalyptus spp |
author |
Fonseca, Natália Risso |
author_facet |
Fonseca, Natália Risso |
author_role |
author |
dc.contributor.authorLattes.por.fl_str_mv |
http://lattes.cnpq.br/2709153501578306 |
dc.contributor.author.fl_str_mv |
Fonseca, Natália Risso |
dc.contributor.advisor-co1.fl_str_mv |
Lopes, Carlos Alberto |
dc.contributor.advisor-co1Lattes.fl_str_mv |
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4783372D6 |
dc.contributor.advisor-co2.fl_str_mv |
Cunha, Luis Claudio Vieira da |
dc.contributor.advisor-co2Lattes.fl_str_mv |
http://lattes.cnpq.br/9367947173322278 |
dc.contributor.advisor1.fl_str_mv |
Alfenas, Acelino Couto |
dc.contributor.advisor1Lattes.fl_str_mv |
http://lattes.cnpq.br/2514320654462590 |
dc.contributor.referee1.fl_str_mv |
Guimarães, Lúcio Mauro da Silva |
dc.contributor.referee1Lattes.fl_str_mv |
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4766939H1 |
contributor_str_mv |
Lopes, Carlos Alberto Cunha, Luis Claudio Vieira da Alfenas, Acelino Couto Guimarães, Lúcio Mauro da Silva |
dc.subject.por.fl_str_mv |
Murcha bacteriana Ralstonia solanacearum Eucalipto Resistência |
topic |
Murcha bacteriana Ralstonia solanacearum Eucalipto Resistência Bacterial wilt Ralstonia solanacearum Eucalyptus Resistance CNPQ::CIENCIAS AGRARIAS::AGRONOMIA::FITOSSANIDADE::FITOPATOLOGIA |
dc.subject.eng.fl_str_mv |
Bacterial wilt Ralstonia solanacearum Eucalyptus Resistance |
dc.subject.cnpq.fl_str_mv |
CNPQ::CIENCIAS AGRARIAS::AGRONOMIA::FITOSSANIDADE::FITOPATOLOGIA |
description |
The Ralstonia wilt of eucalyptus, caused by Ralstonia solanacearum, is potentially one of the major disease of culture. Because of its high genetic and phenotypic variability, currently R. solanacearum is considered a "species complex", subdivided into four taxonomic levels: species, filotipo, sequevar and clone. In order to understand the variability among isolates of R. solanacearum from eucalyptus and develop a protocol for inoculation and screening of resistant clones of eucalyptus to Ralstonia wilt this work was performed. The work was divided into two articles: (i) Molecular characterization of isolates of Ralstonia solanacearum in Eucalyptus spp. and (ii) Method of inoculation and evaluation of resistant clones of Eucalyptus spp. to Ralstonia wilt caused by Ralstonia solanacearum. In Article 1, 19 isolates were analyzed from different regions of eucalyptus in Brazil based on new classification (filotipo and sequevar) and its genetic variability. A product of 372 bp generated by multiplex-PCR amplification using primers Nmult permitted the identification of all isolates analyzed as pertaining to filotipo II. Eighteen isolates were grouped in subclade IIA and just one isolate in IIB. The phylogenetic tree generated from the endoglucanase (egl) gene sequences confirmed the classification of the isolates in filotipo II and separated them into sequevares. The strains AMC22, IBSBF2568 and IBSBF2576 were grouped into a single clade, as well as isolates UFV18 and UFV20, with 89% and 78% posterior probability, respectively, forming two new possible sequevares not defined yet. We also identified isolates belonging to sequevar 41 (100% probability) and 37 (88% probability). However, most of the isolates did not fit in any of previously described sequevar, or formed defined clades. The analysis of results of obtained fragments amplified by ERIC-PCR technique suggested a great genetic diversity among the isolates analyzed in this work. In addition, a high correlation between the geographical origin of isolates and the similarity showed by them was observed. In Article 2, three methods of inoculation and resistant of Eucalyptus spp. clones to Ralstonia wilt was evaluated: (i) soil infestation with R. solanacearum, (ii) dipping of sectioned roots in bacterial suspension, and (iii) injection of a bacterial suspension in the base of the stem. The injection method of a bacterial suspension at the stem base was the most efficient inoculation method of R. solanacearum on eucalyptus. Four, out of 21 clones tested for Ralstonia wilt resistance by this method, were classified as resistant by not showing symptoms of wilt and bacterial exudation up to 30 days after inoculation. |
publishDate |
2012 |
dc.date.issued.fl_str_mv |
2012-07-18 |
dc.date.available.fl_str_mv |
2013-04-23 2015-03-26T13:37:49Z |
dc.date.accessioned.fl_str_mv |
2015-03-26T13:37:49Z |
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info:eu-repo/semantics/publishedVersion |
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info:eu-repo/semantics/masterThesis |
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masterThesis |
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publishedVersion |
dc.identifier.citation.fl_str_mv |
FONSECA, Natália Risso. Molecular characterization and inoculation of Ralstonia solanacearum in Eucalyptus spp. 2012. 54 f. Dissertação (Mestrado em Etiologia; Epidemiologia; Controle) - Universidade Federal de Viçosa, Viçosa, 2012. |
dc.identifier.uri.fl_str_mv |
http://locus.ufv.br/handle/123456789/4412 |
identifier_str_mv |
FONSECA, Natália Risso. Molecular characterization and inoculation of Ralstonia solanacearum in Eucalyptus spp. 2012. 54 f. Dissertação (Mestrado em Etiologia; Epidemiologia; Controle) - Universidade Federal de Viçosa, Viçosa, 2012. |
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http://locus.ufv.br/handle/123456789/4412 |
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Universidade Federal de Viçosa |
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Etiologia; Epidemiologia; Controle |
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Universidade Federal de Viçosa |
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