Sirosporium diffusum associado à nogueira-pecã: patogenicidade, morfofisiologia, filogenia e escala diagramática
Autor(a) principal: | |
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Data de Publicação: | 2018 |
Tipo de documento: | Dissertação |
Idioma: | por |
Título da fonte: | Manancial - Repositório Digital da UFSM |
dARK ID: | ark:/26339/001300000157x |
Texto Completo: | http://repositorio.ufsm.br/handle/1/19175 |
Resumo: | The culture of the pecan [Carya illinoinensis (Wangenh.) K. Koch], family Juglandaceae, have economic importance in southern Brazil and cultivation grows rapidly. Consequently, the increase of planted area has triggered frequent epidemics, mainly of fungal diseases, which are favored by the climate and the accumulation of susceptible hosts. Recently, a new disease was diagnosed and reported in Brazilian plantations [(POLETTO et al., 2017), https://doi.org/10.1094/PDIS-06-16-0820-PDN], which is caused by fungi Sirosporium diffusum. The disease causes defoliation of the plants, reducing the production of fruits. In view of this observation, the objective of this work was to evaluate the pathogenicity of isolates of Sirosporium sp. associated with brown leaf spot of the pecan, characterize them morphological, physiological, and molecular, as well as to develop a disease diagrammatic scale for epidemiological studies. For that, samples of symptomatic leaves were collected in 10 pecan orchards, in the state of Rio Grande do Sul. After obtaining the 10 isolates of the pathogen, a pathogenicity test was conducted for the evaluation of the incidence and virulence of the pathogen, characterized morphologically and the variables used in the UPGMA (Method of grouping the mean distance) analysis in which the isolates were grouped in a dendrogram according to the morphological dissimilarity and the characters that contributed the most to the divergence were established. Molecular analysis was performed by amplifying and sequencing the ITS and tef1- regions of the DNA. For physiological characterization, eight culture media and five light regimes were combined in the first assay, while in the second, the three best treatments from the first assay which conferred higher sporulation were combined with three temperatures. For the elaboration of the diagrammatic scale, 240 symptomatic leaflets were collected, from which the real severity of the disease was obtained in percentage terms. For the construction of the scale, the observed limits of minimum and maximum disease severity plus six intermediate levels were used. The validation was performed by eight evaluators, first without and, subsequently, with the use of the scale. The results showed through the pathogenicity test that all isolates were pathogenic and the incidence was 100% for most of them, with differences in the virulence capacity. The phylogenetic analysis confirmed the isolates as Sirosporium diffusum, the causative agent of the brown leaf spot in C. illinoinensis in Brazil. Sequencing of the ITS region and the tef1- gene were effective for identification of the fungus as S. diffusum. The best artificial conditions of culture in which S. diffusum shows good mycelial growth and sporulation together was obtained with photoperiod of 24 hour, temperature of 20 ±1°C and culture media V8CaCO3 or CA. The diagrammatic scale proposed for evaluation of brown leaf spot in C. illinoinensis provides good accuracy, precision and reproducibility of the estimates to the evaluators, besides being easy to use, and epidemiological studies of the disease are adequate. |
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Sirosporium diffusum associado à nogueira-pecã: patogenicidade, morfofisiologia, filogenia e escala diagramáticaSirosporium diffusum associated to pecan: pathogenicity, morphophysiology, phylogeny and diagramatic scaleCarya illinoinensisMancha foliar marromSequenciamentoBrown leaf spotSequencingCNPQ::CIENCIAS AGRARIAS::RECURSOS FLORESTAIS E ENGENHARIA FLORESTALThe culture of the pecan [Carya illinoinensis (Wangenh.) K. Koch], family Juglandaceae, have economic importance in southern Brazil and cultivation grows rapidly. Consequently, the increase of planted area has triggered frequent epidemics, mainly of fungal diseases, which are favored by the climate and the accumulation of susceptible hosts. Recently, a new disease was diagnosed and reported in Brazilian plantations [(POLETTO et al., 2017), https://doi.org/10.1094/PDIS-06-16-0820-PDN], which is caused by fungi Sirosporium diffusum. The disease causes defoliation of the plants, reducing the production of fruits. In view of this observation, the objective of this work was to evaluate the pathogenicity of isolates of Sirosporium sp. associated with brown leaf spot of the pecan, characterize them morphological, physiological, and molecular, as well as to develop a disease diagrammatic scale for epidemiological studies. For that, samples of symptomatic leaves were collected in 10 pecan orchards, in the state of Rio Grande do Sul. After obtaining the 10 isolates of the pathogen, a pathogenicity test was conducted for the evaluation of the incidence and virulence of the pathogen, characterized morphologically and the variables used in the UPGMA (Method of grouping the mean distance) analysis in which the isolates were grouped in a dendrogram according to the morphological dissimilarity and the characters that contributed the most to the divergence were established. Molecular analysis was performed by amplifying and sequencing the ITS and tef1- regions of the DNA. For physiological characterization, eight culture media and five light regimes were combined in the first assay, while in the second, the three best treatments from the first assay which conferred higher sporulation were combined with three temperatures. For the elaboration of the diagrammatic scale, 240 symptomatic leaflets were collected, from which the real severity of the disease was obtained in percentage terms. For the construction of the scale, the observed limits of minimum and maximum disease severity plus six intermediate levels were used. The validation was performed by eight evaluators, first without and, subsequently, with the use of the scale. The results showed through the pathogenicity test that all isolates were pathogenic and the incidence was 100% for most of them, with differences in the virulence capacity. The phylogenetic analysis confirmed the isolates as Sirosporium diffusum, the causative agent of the brown leaf spot in C. illinoinensis in Brazil. Sequencing of the ITS region and the tef1- gene were effective for identification of the fungus as S. diffusum. The best artificial conditions of culture in which S. diffusum shows good mycelial growth and sporulation together was obtained with photoperiod of 24 hour, temperature of 20 ±1°C and culture media V8CaCO3 or CA. The diagrammatic scale proposed for evaluation of brown leaf spot in C. illinoinensis provides good accuracy, precision and reproducibility of the estimates to the evaluators, besides being easy to use, and epidemiological studies of the disease are adequate.Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPESA cultura da nogueira-pecã [Carya illinoinensis (Wangenh.) K. Koch], família Juglandaceae, tem importância econômica no sul do Brasil e o cultivo cresce expressivamente a cada ano. Por conseguinte, o aumento da área plantada tem desencadeado frequentes epidemias, principalmente de doenças fúngicas, as quais são favorecidas pelo clima úmido e acúmulo de hospedeiros suscetíveis. Recentemente, uma nova doença foi diagnosticada e relatada nos plantios brasileiros [(POLETTO et al., 2017), https://doi.org/10.1094/PDIS-06-16-0820-PDN], a qual é causada pelo fungo Sirosporium diffusum. A doença causa desfolha das plantas, prejudicando a produção de frutos. Diante dessa observação, o objetivo deste trabalho foi avaliar a patogenicidade de isolados de Sirosporium sp. associados a mancha foliar marrom da nogueira-pecã, caracterizá-los morfológica, fisiológica e molecularmente, bem como, desenvolver uma escala diagramática para avaliar esta doença em estudos epidemiológicos. Para tanto, foram realizadas coletas de folhas sintomáticas em 10 pomares de nogueira-pecã, no estado do Rio Grande do Sul. Após a obtenção de 10 isolados do patógeno, um teste de patogenicidade foi conduzido para a avaliação da incidência e virulência do patógeno. Os isolados também foram caracterizados morfologicamente, sendo as variáveis utilizadas na análise UPGMA (Método de Agrupamento da Distância Média) em que os isolados foram agrupados em um dendrograma de acordo com a dissimilaridade morfológica e estabelecidos os caracteres que mais contribuíram para a divergência. A análise molecular foi realizada através da amplificação e sequenciamento das regiões ITS e tef1- do DNA. Para a caracterização fisiológica, num primeiro ensaio, combinaram-se oito meios de cultura e cinco regimes de luz, enquanto que no segundo, os três melhores tratamentos do ensaio anterior, que conferiram maior esporulação, foram combinados com três temperaturas. Para a elaboração da escala diagramática foram coletados 240 folíolos sintomáticos, a partir dos quais obteve-se a severidade real da doença em termos percentu-ais. Para a construção da escala, utilizaram-se os limites observados de severidade mínima e máxima da doença, mais seis níveis intermediários. A validação foi realizada por oito avaliadores, primeiramente sem o uso da escala e, na sequência, utilizando-a. Os resultados mostraram através do teste de patogenicidade, que todos os isolados foram patogênicos e a incidência foi de 100% para maioria deles. No entanto, houve diferenças na capacidade de virulência. A análise filogenética confirmou Sirosporium diffusum como o agente causador da mancha foliar marrom em C. illinoinensis no Brasil. O sequenciamento da região ITS e do gene tef1- foram eficazes para a identificação do fungo como S. diffusum. As melhores condições artificiais de cultivo em que S. diffusum apresenta bom crescimento micelial e esporulação de forma conjunta foi obtida com fotoperíodo de 24 horas, temperatura de 20 ±1°C e com os meios de cultura V8CaCO3 ou CA. A escala diagramática proposta para avaliação de mancha foliar marrom em C. illinoinensis proporciona boa acurácia, precisão e reprodutibilidade das estimativas aos avaliadores, além de ser de fácil utilização, sendo adequada para estudos epidemiológicos da doença.Universidade Federal de Santa MariaBrasilRecursos Florestais e Engenharia FlorestalUFSMPrograma de Pós-Graduação em Engenharia FlorestalCentro de Ciências RuraisMuniz, Marlove Fatima Briãohttp://lattes.cnpq.br/3148312031889388Heldwein, Arno Bernardohttp://lattes.cnpq.br/6671155842231311Brioso, Paulo Sergio Torreshttp://lattes.cnpq.br/8099996221105627Poletto, Tales2019-12-18T13:15:07Z2019-12-18T13:15:07Z2018-02-27info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisapplication/pdfhttp://repositorio.ufsm.br/handle/1/19175ark:/26339/001300000157xporAttribution-NonCommercial-NoDerivatives 4.0 Internationalhttp://creativecommons.org/licenses/by-nc-nd/4.0/info:eu-repo/semantics/openAccessreponame:Manancial - Repositório Digital da UFSMinstname:Universidade Federal de Santa Maria (UFSM)instacron:UFSM2022-05-12T14:07:18Zoai:repositorio.ufsm.br:1/19175Biblioteca Digital de Teses e Dissertaçõeshttps://repositorio.ufsm.br/ONGhttps://repositorio.ufsm.br/oai/requestatendimento.sib@ufsm.br||tedebc@gmail.comopendoar:2022-05-12T14:07:18Manancial - Repositório Digital da UFSM - Universidade Federal de Santa Maria (UFSM)false |
dc.title.none.fl_str_mv |
Sirosporium diffusum associado à nogueira-pecã: patogenicidade, morfofisiologia, filogenia e escala diagramática Sirosporium diffusum associated to pecan: pathogenicity, morphophysiology, phylogeny and diagramatic scale |
title |
Sirosporium diffusum associado à nogueira-pecã: patogenicidade, morfofisiologia, filogenia e escala diagramática |
spellingShingle |
Sirosporium diffusum associado à nogueira-pecã: patogenicidade, morfofisiologia, filogenia e escala diagramática Poletto, Tales Carya illinoinensis Mancha foliar marrom Sequenciamento Brown leaf spot Sequencing CNPQ::CIENCIAS AGRARIAS::RECURSOS FLORESTAIS E ENGENHARIA FLORESTAL |
title_short |
Sirosporium diffusum associado à nogueira-pecã: patogenicidade, morfofisiologia, filogenia e escala diagramática |
title_full |
Sirosporium diffusum associado à nogueira-pecã: patogenicidade, morfofisiologia, filogenia e escala diagramática |
title_fullStr |
Sirosporium diffusum associado à nogueira-pecã: patogenicidade, morfofisiologia, filogenia e escala diagramática |
title_full_unstemmed |
Sirosporium diffusum associado à nogueira-pecã: patogenicidade, morfofisiologia, filogenia e escala diagramática |
title_sort |
Sirosporium diffusum associado à nogueira-pecã: patogenicidade, morfofisiologia, filogenia e escala diagramática |
author |
Poletto, Tales |
author_facet |
Poletto, Tales |
author_role |
author |
dc.contributor.none.fl_str_mv |
Muniz, Marlove Fatima Brião http://lattes.cnpq.br/3148312031889388 Heldwein, Arno Bernardo http://lattes.cnpq.br/6671155842231311 Brioso, Paulo Sergio Torres http://lattes.cnpq.br/8099996221105627 |
dc.contributor.author.fl_str_mv |
Poletto, Tales |
dc.subject.por.fl_str_mv |
Carya illinoinensis Mancha foliar marrom Sequenciamento Brown leaf spot Sequencing CNPQ::CIENCIAS AGRARIAS::RECURSOS FLORESTAIS E ENGENHARIA FLORESTAL |
topic |
Carya illinoinensis Mancha foliar marrom Sequenciamento Brown leaf spot Sequencing CNPQ::CIENCIAS AGRARIAS::RECURSOS FLORESTAIS E ENGENHARIA FLORESTAL |
description |
The culture of the pecan [Carya illinoinensis (Wangenh.) K. Koch], family Juglandaceae, have economic importance in southern Brazil and cultivation grows rapidly. Consequently, the increase of planted area has triggered frequent epidemics, mainly of fungal diseases, which are favored by the climate and the accumulation of susceptible hosts. Recently, a new disease was diagnosed and reported in Brazilian plantations [(POLETTO et al., 2017), https://doi.org/10.1094/PDIS-06-16-0820-PDN], which is caused by fungi Sirosporium diffusum. The disease causes defoliation of the plants, reducing the production of fruits. In view of this observation, the objective of this work was to evaluate the pathogenicity of isolates of Sirosporium sp. associated with brown leaf spot of the pecan, characterize them morphological, physiological, and molecular, as well as to develop a disease diagrammatic scale for epidemiological studies. For that, samples of symptomatic leaves were collected in 10 pecan orchards, in the state of Rio Grande do Sul. After obtaining the 10 isolates of the pathogen, a pathogenicity test was conducted for the evaluation of the incidence and virulence of the pathogen, characterized morphologically and the variables used in the UPGMA (Method of grouping the mean distance) analysis in which the isolates were grouped in a dendrogram according to the morphological dissimilarity and the characters that contributed the most to the divergence were established. Molecular analysis was performed by amplifying and sequencing the ITS and tef1- regions of the DNA. For physiological characterization, eight culture media and five light regimes were combined in the first assay, while in the second, the three best treatments from the first assay which conferred higher sporulation were combined with three temperatures. For the elaboration of the diagrammatic scale, 240 symptomatic leaflets were collected, from which the real severity of the disease was obtained in percentage terms. For the construction of the scale, the observed limits of minimum and maximum disease severity plus six intermediate levels were used. The validation was performed by eight evaluators, first without and, subsequently, with the use of the scale. The results showed through the pathogenicity test that all isolates were pathogenic and the incidence was 100% for most of them, with differences in the virulence capacity. The phylogenetic analysis confirmed the isolates as Sirosporium diffusum, the causative agent of the brown leaf spot in C. illinoinensis in Brazil. Sequencing of the ITS region and the tef1- gene were effective for identification of the fungus as S. diffusum. The best artificial conditions of culture in which S. diffusum shows good mycelial growth and sporulation together was obtained with photoperiod of 24 hour, temperature of 20 ±1°C and culture media V8CaCO3 or CA. The diagrammatic scale proposed for evaluation of brown leaf spot in C. illinoinensis provides good accuracy, precision and reproducibility of the estimates to the evaluators, besides being easy to use, and epidemiological studies of the disease are adequate. |
publishDate |
2018 |
dc.date.none.fl_str_mv |
2018-02-27 2019-12-18T13:15:07Z 2019-12-18T13:15:07Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/masterThesis |
format |
masterThesis |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://repositorio.ufsm.br/handle/1/19175 |
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ark:/26339/001300000157x |
url |
http://repositorio.ufsm.br/handle/1/19175 |
identifier_str_mv |
ark:/26339/001300000157x |
dc.language.iso.fl_str_mv |
por |
language |
por |
dc.rights.driver.fl_str_mv |
Attribution-NonCommercial-NoDerivatives 4.0 International http://creativecommons.org/licenses/by-nc-nd/4.0/ info:eu-repo/semantics/openAccess |
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Attribution-NonCommercial-NoDerivatives 4.0 International http://creativecommons.org/licenses/by-nc-nd/4.0/ |
eu_rights_str_mv |
openAccess |
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application/pdf |
dc.publisher.none.fl_str_mv |
Universidade Federal de Santa Maria Brasil Recursos Florestais e Engenharia Florestal UFSM Programa de Pós-Graduação em Engenharia Florestal Centro de Ciências Rurais |
publisher.none.fl_str_mv |
Universidade Federal de Santa Maria Brasil Recursos Florestais e Engenharia Florestal UFSM Programa de Pós-Graduação em Engenharia Florestal Centro de Ciências Rurais |
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reponame:Manancial - Repositório Digital da UFSM instname:Universidade Federal de Santa Maria (UFSM) instacron:UFSM |
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Universidade Federal de Santa Maria (UFSM) |
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UFSM |
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UFSM |
reponame_str |
Manancial - Repositório Digital da UFSM |
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Manancial - Repositório Digital da UFSM |
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Manancial - Repositório Digital da UFSM - Universidade Federal de Santa Maria (UFSM) |
repository.mail.fl_str_mv |
atendimento.sib@ufsm.br||tedebc@gmail.com |
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1815172261744738304 |