Caracterização fenotípica e genotípica de isolados de Pseudomonas spp. do tomateiro tutorado no Brasil

Detalhes bibliográficos
Autor(a) principal: Mota, Lara Caroline Borges Moreira Mota
Data de Publicação: 2020
Tipo de documento: Tese
Idioma: por
Título da fonte: Repositório Institucional da UFU
Texto Completo: https://repositorio.ufu.br/handle/123456789/28836
http://doi.org/10.14393/ufu.te.2020.270
Resumo: The bacterial incidence caused by the bacteria genus Pseudomonas cause considerable losses in tomato yield. This genus has population the isolates with great genetic diversity, it can directly influence the pathogen aggressiveness level. The objective of this work was to characterize 43 isolates of Pseudomonas spp. obtained from samples of tomato stems or leaves with symptoms of pith necrosis or leaf lesions, respectively, regarding phenotypic and genotypic aspects. Bacterial isolates were morphological and biochemically characterized by Gram tests, anaerobic growth, King B medium and LOPAT tests. Molecular analysis was performed by rep-PCR (REP, ERIC and BOX primers), specific primers and genetic sequencing of the 16S region of from isolates part. To evaluate aggressiveness, two assays were conducted, one using the isolates from the stem and the another from the tomato leaf. In both experiments healthy tomato plants cv. Santa Clara VF5600 were inoculated with bacterial suspension at a concentration 1.0 x 108 CFU mL-1, and the severity of the disease was evaluated and the Area Under The Disease Progression Curve calculated. Morfologically, all colonies were circular and bright, most then of transparent and with smooth edges. The elevation varied between straight, slightly elevated and convex, and the coloration in shades of straw color. Biochemically, the isolates were classified into groups II, IVb, Va and Vb the LOPAT tests, one isolate as P. corrugata and two biochemically undetermined isolates. Molecularly the isolates shoued high genetic diversity by rep-PCR, and it was not possible to relate the molecular characteristics with the place of origin. With the specific primers only the UFU B39 isolate was identified as P. corrugata. Genetic sequencing confirmed the identified of several isolates such as P. putida and P. fluorescens, and also P. syringae pv. tomato that presented biochemical variation. In the aggressiveness tests in the stem, the isolates UFU B39 and UFU H24 were the most aggressive, and in the leaf test there was no difference in the isolates aggressiveness. It was concluded that the isolates showed low morfological variation, high biochemical and molecular variation and different levels of stem aggressiveness. This is the first report of P. putida and P. fluorescens causing disease in staked tomato plants in Brazil.
id UFU_3ccd711c807fd4f7655db67ae510c971
oai_identifier_str oai:repositorio.ufu.br:123456789/28836
network_acronym_str UFU
network_name_str Repositório Institucional da UFU
repository_id_str
spelling Caracterização fenotípica e genotípica de isolados de Pseudomonas spp. do tomateiro tutorado no BrasilPhenotypic and genotypic characterization of Pseudomonas spp. isolates from staked tomato in BrazilSolanum lycopersicum L.Diversidade genéticaNecrose da medulaPinta bacterianaQueima das folhasCNPQ::CIENCIAS AGRARIAS::AGRONOMIA::FITOSSANIDADE::FITOPATOLOGIAThe bacterial incidence caused by the bacteria genus Pseudomonas cause considerable losses in tomato yield. This genus has population the isolates with great genetic diversity, it can directly influence the pathogen aggressiveness level. The objective of this work was to characterize 43 isolates of Pseudomonas spp. obtained from samples of tomato stems or leaves with symptoms of pith necrosis or leaf lesions, respectively, regarding phenotypic and genotypic aspects. Bacterial isolates were morphological and biochemically characterized by Gram tests, anaerobic growth, King B medium and LOPAT tests. Molecular analysis was performed by rep-PCR (REP, ERIC and BOX primers), specific primers and genetic sequencing of the 16S region of from isolates part. To evaluate aggressiveness, two assays were conducted, one using the isolates from the stem and the another from the tomato leaf. In both experiments healthy tomato plants cv. Santa Clara VF5600 were inoculated with bacterial suspension at a concentration 1.0 x 108 CFU mL-1, and the severity of the disease was evaluated and the Area Under The Disease Progression Curve calculated. Morfologically, all colonies were circular and bright, most then of transparent and with smooth edges. The elevation varied between straight, slightly elevated and convex, and the coloration in shades of straw color. Biochemically, the isolates were classified into groups II, IVb, Va and Vb the LOPAT tests, one isolate as P. corrugata and two biochemically undetermined isolates. Molecularly the isolates shoued high genetic diversity by rep-PCR, and it was not possible to relate the molecular characteristics with the place of origin. With the specific primers only the UFU B39 isolate was identified as P. corrugata. Genetic sequencing confirmed the identified of several isolates such as P. putida and P. fluorescens, and also P. syringae pv. tomato that presented biochemical variation. In the aggressiveness tests in the stem, the isolates UFU B39 and UFU H24 were the most aggressive, and in the leaf test there was no difference in the isolates aggressiveness. It was concluded that the isolates showed low morfological variation, high biochemical and molecular variation and different levels of stem aggressiveness. This is the first report of P. putida and P. fluorescens causing disease in staked tomato plants in Brazil.FAU - Fundação de Apoio UniversitárioTese (Doutorado)A incidência de bacterioses ocasionadas pelas bactérias do gênero Pseudomonas causam perdas consideráveis na produtividade no tomateiro. Esse gênero possui populações de isolados com grande diversidade genética, o que pode influenciar diretamente no nível de agressividade do patógeno. O objetivo deste trabalho foi caracterizar 43 isolados de Pseudomonas spp. obtidos de amostras de hastes ou folhas de tomateiro com sintomas de necrose da medula ou lesões foliares, respectivamente, quanto à aspectos fenotípicps e genotípicos. Os isolados bacterianos foram caracterizados morfológica e bioquimicamente pelos testes de Gram, crescimento anaeróbico, meio King B e testes LOPAT. A análise molecular foi realizada por rep-PCR (iniciadores REP, ERIC e BOX), iniciadores específicos e sequenciamento genético da região 16S de parte dos isolados. Para avaliar a agressividade, foram conduzidos dois experimentos, um com os isolados provenientes da haste e outro da folha do tomateiro. Em ambos os experimentos as plantas sadias de tomate cv. Santa Clara VF5600 foram inoculadas com suspensão bacteriana na concentração 1,0 x 108 UFC mL-1, sendo avaliada a severidade da doença com escala diagramática e calculada a Área Abaixo da Curva de Progresso da Doença. Quanto à morfologia, todas as colônias foram circulares e brilhantes, a maioria transparente e com bordas lisas. A elevação variou entre reta, levemente elevada e convexa, e a coloração em nuances de cor palha. Bioquimicamente, os isolados foram classificados nos grupos II, IVb, Va e Vb dos testes LOPAT, um isolado como P. corrugata e dois isolados bioquimicamente não determinados. Molecularmente, apresentaram grande diversidade genética por rep-PCR, não sendo possível relacionar as características moleculares com o local de origem. Com os iniciadores específicos apenas o isolado UFU B39 foi identificado como P. corrugata. No sequenciamento genético foi possível confirmar a identidade de vários isolados como sendo P. putida e P. fluorescens, bem como P. syringae pv. tomato, os quais apresentaram variação bioquímica. Nos testes de agressividade na haste os isolados UFU B39 e UFU H24 foram os mais agressivos, enquanto nos testes de folha não foi observada diferença na agressividade dos isolados. Dessa forma, conclui-se que os isolados apresentaram pouca variação morfológica, muita variação bioquímica e molecular e diferentes níveis de agressividade na haste. Este é o primeiro relato de P. putida e P. fluorescens causando doença no tomateiro tutorado no Brasil.2022-02-21Universidade Federal de UberlândiaBrasilPrograma de Pós-graduação em AgronomiaTebaldi, Nilvanira Donizetehttp://lattes.cnpq.br/0695770301548519Luz, José Magno Queirozhttp://lattes.cnpq.br/7022519480996266Lima, Alison Talis Martinshttp://lattes.cnpq.br/5848428341840005Pereira, Igor Souzahttp://lattes.cnpq.br/0610814748801550Duval, Alice Maria Quezadohttp://lattes.cnpq.br/3456750603463387Mota, Lara Caroline Borges Moreira Mota2020-02-27T13:03:43Z2020-02-27T13:03:43Z2020-02-21info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesisapplication/pdfMOTA, Lara Caroline Borges Moreira. Caracterização fenotípica e genotípica de isolados de Pseudomonas spp. do tomateiro tutorado no Brasil. 2020. 148 f. Tese. (Doutorado em Agronomia /Fitotecnia) - Universidade Federal de Uberlândia, Uberlândia, 2020. DOI http://doi.org/10.14393/ufu.te.2020.270.https://repositorio.ufu.br/handle/123456789/28836http://doi.org/10.14393/ufu.te.2020.270porAttribution-NonCommercial-NoDerivs 3.0 United Stateshttp://creativecommons.org/licenses/by-nc-nd/3.0/us/info:eu-repo/semantics/openAccessreponame:Repositório Institucional da UFUinstname:Universidade Federal de Uberlândia (UFU)instacron:UFU2022-10-17T19:07:11Zoai:repositorio.ufu.br:123456789/28836Repositório InstitucionalONGhttp://repositorio.ufu.br/oai/requestdiinf@dirbi.ufu.bropendoar:2022-10-17T19:07:11Repositório Institucional da UFU - Universidade Federal de Uberlândia (UFU)false
dc.title.none.fl_str_mv Caracterização fenotípica e genotípica de isolados de Pseudomonas spp. do tomateiro tutorado no Brasil
Phenotypic and genotypic characterization of Pseudomonas spp. isolates from staked tomato in Brazil
title Caracterização fenotípica e genotípica de isolados de Pseudomonas spp. do tomateiro tutorado no Brasil
spellingShingle Caracterização fenotípica e genotípica de isolados de Pseudomonas spp. do tomateiro tutorado no Brasil
Mota, Lara Caroline Borges Moreira Mota
Solanum lycopersicum L.
Diversidade genética
Necrose da medula
Pinta bacteriana
Queima das folhas
CNPQ::CIENCIAS AGRARIAS::AGRONOMIA::FITOSSANIDADE::FITOPATOLOGIA
title_short Caracterização fenotípica e genotípica de isolados de Pseudomonas spp. do tomateiro tutorado no Brasil
title_full Caracterização fenotípica e genotípica de isolados de Pseudomonas spp. do tomateiro tutorado no Brasil
title_fullStr Caracterização fenotípica e genotípica de isolados de Pseudomonas spp. do tomateiro tutorado no Brasil
title_full_unstemmed Caracterização fenotípica e genotípica de isolados de Pseudomonas spp. do tomateiro tutorado no Brasil
title_sort Caracterização fenotípica e genotípica de isolados de Pseudomonas spp. do tomateiro tutorado no Brasil
author Mota, Lara Caroline Borges Moreira Mota
author_facet Mota, Lara Caroline Borges Moreira Mota
author_role author
dc.contributor.none.fl_str_mv Tebaldi, Nilvanira Donizete
http://lattes.cnpq.br/0695770301548519
Luz, José Magno Queiroz
http://lattes.cnpq.br/7022519480996266
Lima, Alison Talis Martins
http://lattes.cnpq.br/5848428341840005
Pereira, Igor Souza
http://lattes.cnpq.br/0610814748801550
Duval, Alice Maria Quezado
http://lattes.cnpq.br/3456750603463387
dc.contributor.author.fl_str_mv Mota, Lara Caroline Borges Moreira Mota
dc.subject.por.fl_str_mv Solanum lycopersicum L.
Diversidade genética
Necrose da medula
Pinta bacteriana
Queima das folhas
CNPQ::CIENCIAS AGRARIAS::AGRONOMIA::FITOSSANIDADE::FITOPATOLOGIA
topic Solanum lycopersicum L.
Diversidade genética
Necrose da medula
Pinta bacteriana
Queima das folhas
CNPQ::CIENCIAS AGRARIAS::AGRONOMIA::FITOSSANIDADE::FITOPATOLOGIA
description The bacterial incidence caused by the bacteria genus Pseudomonas cause considerable losses in tomato yield. This genus has population the isolates with great genetic diversity, it can directly influence the pathogen aggressiveness level. The objective of this work was to characterize 43 isolates of Pseudomonas spp. obtained from samples of tomato stems or leaves with symptoms of pith necrosis or leaf lesions, respectively, regarding phenotypic and genotypic aspects. Bacterial isolates were morphological and biochemically characterized by Gram tests, anaerobic growth, King B medium and LOPAT tests. Molecular analysis was performed by rep-PCR (REP, ERIC and BOX primers), specific primers and genetic sequencing of the 16S region of from isolates part. To evaluate aggressiveness, two assays were conducted, one using the isolates from the stem and the another from the tomato leaf. In both experiments healthy tomato plants cv. Santa Clara VF5600 were inoculated with bacterial suspension at a concentration 1.0 x 108 CFU mL-1, and the severity of the disease was evaluated and the Area Under The Disease Progression Curve calculated. Morfologically, all colonies were circular and bright, most then of transparent and with smooth edges. The elevation varied between straight, slightly elevated and convex, and the coloration in shades of straw color. Biochemically, the isolates were classified into groups II, IVb, Va and Vb the LOPAT tests, one isolate as P. corrugata and two biochemically undetermined isolates. Molecularly the isolates shoued high genetic diversity by rep-PCR, and it was not possible to relate the molecular characteristics with the place of origin. With the specific primers only the UFU B39 isolate was identified as P. corrugata. Genetic sequencing confirmed the identified of several isolates such as P. putida and P. fluorescens, and also P. syringae pv. tomato that presented biochemical variation. In the aggressiveness tests in the stem, the isolates UFU B39 and UFU H24 were the most aggressive, and in the leaf test there was no difference in the isolates aggressiveness. It was concluded that the isolates showed low morfological variation, high biochemical and molecular variation and different levels of stem aggressiveness. This is the first report of P. putida and P. fluorescens causing disease in staked tomato plants in Brazil.
publishDate 2020
dc.date.none.fl_str_mv 2020-02-27T13:03:43Z
2020-02-27T13:03:43Z
2020-02-21
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.uri.fl_str_mv MOTA, Lara Caroline Borges Moreira. Caracterização fenotípica e genotípica de isolados de Pseudomonas spp. do tomateiro tutorado no Brasil. 2020. 148 f. Tese. (Doutorado em Agronomia /Fitotecnia) - Universidade Federal de Uberlândia, Uberlândia, 2020. DOI http://doi.org/10.14393/ufu.te.2020.270.
https://repositorio.ufu.br/handle/123456789/28836
http://doi.org/10.14393/ufu.te.2020.270
identifier_str_mv MOTA, Lara Caroline Borges Moreira. Caracterização fenotípica e genotípica de isolados de Pseudomonas spp. do tomateiro tutorado no Brasil. 2020. 148 f. Tese. (Doutorado em Agronomia /Fitotecnia) - Universidade Federal de Uberlândia, Uberlândia, 2020. DOI http://doi.org/10.14393/ufu.te.2020.270.
url https://repositorio.ufu.br/handle/123456789/28836
http://doi.org/10.14393/ufu.te.2020.270
dc.language.iso.fl_str_mv por
language por
dc.rights.driver.fl_str_mv Attribution-NonCommercial-NoDerivs 3.0 United States
http://creativecommons.org/licenses/by-nc-nd/3.0/us/
info:eu-repo/semantics/openAccess
rights_invalid_str_mv Attribution-NonCommercial-NoDerivs 3.0 United States
http://creativecommons.org/licenses/by-nc-nd/3.0/us/
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Universidade Federal de Uberlândia
Brasil
Programa de Pós-graduação em Agronomia
publisher.none.fl_str_mv Universidade Federal de Uberlândia
Brasil
Programa de Pós-graduação em Agronomia
dc.source.none.fl_str_mv reponame:Repositório Institucional da UFU
instname:Universidade Federal de Uberlândia (UFU)
instacron:UFU
instname_str Universidade Federal de Uberlândia (UFU)
instacron_str UFU
institution UFU
reponame_str Repositório Institucional da UFU
collection Repositório Institucional da UFU
repository.name.fl_str_mv Repositório Institucional da UFU - Universidade Federal de Uberlândia (UFU)
repository.mail.fl_str_mv diinf@dirbi.ufu.br
_version_ 1813711591240105984