Sirosporium diffusum associado à nogueira-pecã: patogenicidade, morfofisiologia, filogenia e escala diagramática

Detalhes bibliográficos
Autor(a) principal: Poletto, Tales
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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spelling 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
dc.identifier.dark.fl_str_mv 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
rights_invalid_str_mv Attribution-NonCommercial-NoDerivatives 4.0 International
http://creativecommons.org/licenses/by-nc-nd/4.0/
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv 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
dc.source.none.fl_str_mv reponame:Manancial - Repositório Digital da UFSM
instname:Universidade Federal de Santa Maria (UFSM)
instacron:UFSM
instname_str Universidade Federal de Santa Maria (UFSM)
instacron_str UFSM
institution UFSM
reponame_str Manancial - Repositório Digital da UFSM
collection Manancial - Repositório Digital da UFSM
repository.name.fl_str_mv 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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