Identificação, caracterização e controle in vitro de Phoma pedeiae (Aveskamp, Gruyter & Verkley) associado à Cedrela fissilis Vell.

Detalhes bibliográficos
Autor(a) principal: Dutra, Adriana Falcão
Data de Publicação: 2018
Tipo de documento: Tese
Idioma: por
Título da fonte: Manancial - Repositório Digital da UFSM
Texto Completo: http://repositorio.ufsm.br/handle/1/15107
Resumo: The species Cedrela fissilis Vell. (cedro) presents good growth, producing wood of excellent quality, causing intense exploitation by the loggers. One of the main problems that compromise the production of wood is the occurrence of pests and diseases, causing serious damages to the host and changes in the physiological processes, leading to the death of the plant. Thus, the study of the biological activity of essential oils (OEs) of plants can constitute a potential form of control of diseases in plants. The objective of this study was the identification and morphological and molecular characterization of Phoma spp. associated with Cedrela fissilis Vell., and to evaluate the antifungal activity of cedro OE in the in vitro control of Phoma pedeiae. Leaflets were collected from adult trees with symptoms of the disease (Santa Maria, RS) and sent to the Elocy Minussi Phytopathology Laboratory (UFSM) for isolation of the fungus. The material was observed on stereoscopic and optical microscopes, and when fungal structures were present, they were transferred to Petri dishes containing BDA. After 14 days of development, the fungus was purified by monosporic culture and transferred to the BDA, CA and V8 media for morphological and molecular characterization. The pathogenicity of the fungus was performed by sprinkling on leaves for two isolates. The morphological characteristics analyzed were: mycelial growth (MG), mycelial growth rate (MGR) and growth rate index (GRI), sporulation, colony pigmentation and conidia characterization. OE extraction was carried out in the Laboratory of Extractive Plants, and OE was extracted from young cedro leaflets, in the spring season of 2015, 2016 and 2017, by hydrodistillation. The in vitro antifungal activity of leaflets OE was on pure colonies of Phoma pedeiae, at concentrations of 0.5, 0.75, 1.0 and 1.5 μL mL-1, in addition to positive and negative controls. Daily measurements of the colony diameter were carried out for 14 days, with MG, MGR, GRI and inhibition of mycelial growth. Sequences were deposited in GenBank and 70 and 81% similarly, with the fungus Didymella pedeiae, sexual form of Phoma pedeiae, by means of the sequencing of genes from the ITS and β-tubulin regions. The symptoms caused by the fungus started at 20 days after inoculation, not differing between the two isolates, causing the death of the individuals in 50 days. The fungus presented rounded pycnidia of black coloration, white mycelium and oblong oval conidia, without septum and hyaline. Sporulation in the V8 culture medium was superior to the BDA and CA medium. At the end of the three-year evaluation, 45 OE components of cedar leaflets were identified, corresponding to approximately 90% of the total OE composition, with the predominance of β-caryophyllene, spathulenol and caryophyllene oxide. The cedro OE presented relative antifungal activity against P. pedeiae starting at the concentration of 0.5 μL mL-1. There fore, it can be indicated for use in the control of pathogens, because it is a secondary metabolite, not causing damage to the environment.
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spelling Identificação, caracterização e controle in vitro de Phoma pedeiae (Aveskamp, Gruyter & Verkley) associado à Cedrela fissilis Vell.Identification, characterization and in vitro control of Phoma pedeiae (Aveskamp, Gruyter & Verkley) associated with Cedrela fissilis Vell.FitopatologiaControle de doençasÓleos essenciaisPhytopathologyDisease controlEssencial oilsCNPQ::CIENCIAS AGRARIAS::RECURSOS FLORESTAIS E ENGENHARIA FLORESTALThe species Cedrela fissilis Vell. (cedro) presents good growth, producing wood of excellent quality, causing intense exploitation by the loggers. One of the main problems that compromise the production of wood is the occurrence of pests and diseases, causing serious damages to the host and changes in the physiological processes, leading to the death of the plant. Thus, the study of the biological activity of essential oils (OEs) of plants can constitute a potential form of control of diseases in plants. The objective of this study was the identification and morphological and molecular characterization of Phoma spp. associated with Cedrela fissilis Vell., and to evaluate the antifungal activity of cedro OE in the in vitro control of Phoma pedeiae. Leaflets were collected from adult trees with symptoms of the disease (Santa Maria, RS) and sent to the Elocy Minussi Phytopathology Laboratory (UFSM) for isolation of the fungus. The material was observed on stereoscopic and optical microscopes, and when fungal structures were present, they were transferred to Petri dishes containing BDA. After 14 days of development, the fungus was purified by monosporic culture and transferred to the BDA, CA and V8 media for morphological and molecular characterization. The pathogenicity of the fungus was performed by sprinkling on leaves for two isolates. The morphological characteristics analyzed were: mycelial growth (MG), mycelial growth rate (MGR) and growth rate index (GRI), sporulation, colony pigmentation and conidia characterization. OE extraction was carried out in the Laboratory of Extractive Plants, and OE was extracted from young cedro leaflets, in the spring season of 2015, 2016 and 2017, by hydrodistillation. The in vitro antifungal activity of leaflets OE was on pure colonies of Phoma pedeiae, at concentrations of 0.5, 0.75, 1.0 and 1.5 μL mL-1, in addition to positive and negative controls. Daily measurements of the colony diameter were carried out for 14 days, with MG, MGR, GRI and inhibition of mycelial growth. Sequences were deposited in GenBank and 70 and 81% similarly, with the fungus Didymella pedeiae, sexual form of Phoma pedeiae, by means of the sequencing of genes from the ITS and β-tubulin regions. The symptoms caused by the fungus started at 20 days after inoculation, not differing between the two isolates, causing the death of the individuals in 50 days. The fungus presented rounded pycnidia of black coloration, white mycelium and oblong oval conidia, without septum and hyaline. Sporulation in the V8 culture medium was superior to the BDA and CA medium. At the end of the three-year evaluation, 45 OE components of cedar leaflets were identified, corresponding to approximately 90% of the total OE composition, with the predominance of β-caryophyllene, spathulenol and caryophyllene oxide. The cedro OE presented relative antifungal activity against P. pedeiae starting at the concentration of 0.5 μL mL-1. There fore, it can be indicated for use in the control of pathogens, because it is a secondary metabolite, not causing damage to the environment.Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPESA espécie Cedrela fissilis Vell. (cedro) apresenta bom crescimento, produzindo madeira de excelente qualidade, causando intensa exploração pelos madeireiros. Um dos principais problemas que comprometem a produção de madeira é a ocorrência de pragas e doenças, causando sérios danos ao hospedeiro e alterações nos processos fisiológicos, levando à morte da planta. Assim, o estudo da atividade biológica de óleos essenciais (OEs) de vegetais pode se constituir em uma forma potencial de controle de doenças em plantas. O objetivo deste estudo foi a identificação e caracterização morfofisiológica e molecular de Phoma spp. associado à Cedrela fissilis Vell., e avaliar a atividade antifúngica do OE de cedro no controle in vitro de Phoma pedeiae. Foram coletados folíolos de árvores adultas com sintomas da doença (Santa Maria, RS) e encaminhados ao Laboratório de Fitopatologia Elocy Minussi (UFSM), para isolamento do fungo. O material foi observado em microscópios estereoscópico e ótico, e quando estruturas fúngicas se faziam presentes, eram transferidas para placas de Petri contendo BDA. Após 14 dias de desenvolvimento, o fungo foi purificado por meio da cultura monospórica e transferido para os meios BDA, CA e V8, para caracterização morfofisiológica e molecular. A patogenicidade do fungo foi realizada por meio de aspersão em folhas. As características morfofisiológicas analisadas foram: crescimento micelial (CM), taxa de crescimento micelial (TCM) e índice de velocidade de crescimento (IVCM), esporulação, pigmentação das colônias e caracterização dos conídios. A extração do OE foi conduzida no Laboratório de Extrativos Vegetais, sendo extraído o OE de folíolos jovens de cedro, na estação primaveril dos anos de 2015, 2016 e 2017, por hidrodestilação. A atividade antifúngica in vitro do OE dos folíolos foi a partir de colônias puras de Phoma pedeiae, nas concentrações de 0,5, 0,75, 1,0 e 1,5 μL mL-1, além de controle positivo e negativo. Foram realizadas medições diárias do diâmetro da colônia, por 14 dias, sendo obtido o CM, TCM, IVCM e inibição do crescimento micelial. . A partir da caracterização molecular, por meio do sequenciamento de genes das regiões ITS e β-tubulina, as sequências foram depositadas no GenBank e tiveram similaridade de 70 e 81%, respectivamente, com o fungo Didymella pedeiae, forma sexuada de Phoma pedeiae. Os sintomas causados pelo fungo iniciaram aos 20 dias após a inoculação, não diferindo entre os dois isolados, causando a morte dos indivíduos em 50 dias. O fungo apresentou picnídios arredondados de coloração preta, micélio branco e conídios ovais oblongos, sem septo e hialinos. A esporulação no meio de cultura V8 foi superior ao meio BDA e CA. Ao final dos três anos de avaliação foram identificados 45 componentes do OE de folíolos de cedro, correspondendo a, aproximadamente, 90% da composição total do OE, com a predominância das substâncias β-cariofileno, espatulenol e óxido de cariofileno. O OE de cedro apresentou atividade antifungistática relativa frente a P. pedeiae a partir da concentração de 0,5 μL mL-1. Dessa forma, pode ser indicado para uso no controle de patógenos, por se tratar de um metabólito secundário, não causando prejuízos ao meio ambiente.Universidade Federal de Santa MariaBrasilRecursos Florestais e Engenharia FlorestalUFSMPrograma de Pós-Graduação em Engenharia FlorestalCentro de Ciências RuraisMuniz, Marlove Fatima Briãohttp://lattes.cnpq.br/3148312031889388Oliveira, Andreia Mara Rotta dehttp://lattes.cnpq.br/4046790504087569Blume, Elenahttp://lattes.cnpq.br/3299996746493028Steffen, Gerusa Pauli Kisthttp://lattes.cnpq.br/6467625663365353Kulczynski, Stela Marishttp://lattes.cnpq.br/0604511258822899Dutra, Adriana Falcão2018-12-14T17:05:39Z2018-12-14T17:05:39Z2018-02-26info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesisapplication/pdfhttp://repositorio.ufsm.br/handle/1/15107porAttribution-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:UFSM2018-12-15T05:02:08Zoai:repositorio.ufsm.br:1/15107Biblioteca Digital de Teses e Dissertaçõeshttps://repositorio.ufsm.br/ONGhttps://repositorio.ufsm.br/oai/requestatendimento.sib@ufsm.br||tedebc@gmail.comopendoar:2018-12-15T05:02:08Manancial - Repositório Digital da UFSM - Universidade Federal de Santa Maria (UFSM)false
dc.title.none.fl_str_mv Identificação, caracterização e controle in vitro de Phoma pedeiae (Aveskamp, Gruyter & Verkley) associado à Cedrela fissilis Vell.
Identification, characterization and in vitro control of Phoma pedeiae (Aveskamp, Gruyter & Verkley) associated with Cedrela fissilis Vell.
title Identificação, caracterização e controle in vitro de Phoma pedeiae (Aveskamp, Gruyter & Verkley) associado à Cedrela fissilis Vell.
spellingShingle Identificação, caracterização e controle in vitro de Phoma pedeiae (Aveskamp, Gruyter & Verkley) associado à Cedrela fissilis Vell.
Dutra, Adriana Falcão
Fitopatologia
Controle de doenças
Óleos essenciais
Phytopathology
Disease control
Essencial oils
CNPQ::CIENCIAS AGRARIAS::RECURSOS FLORESTAIS E ENGENHARIA FLORESTAL
title_short Identificação, caracterização e controle in vitro de Phoma pedeiae (Aveskamp, Gruyter & Verkley) associado à Cedrela fissilis Vell.
title_full Identificação, caracterização e controle in vitro de Phoma pedeiae (Aveskamp, Gruyter & Verkley) associado à Cedrela fissilis Vell.
title_fullStr Identificação, caracterização e controle in vitro de Phoma pedeiae (Aveskamp, Gruyter & Verkley) associado à Cedrela fissilis Vell.
title_full_unstemmed Identificação, caracterização e controle in vitro de Phoma pedeiae (Aveskamp, Gruyter & Verkley) associado à Cedrela fissilis Vell.
title_sort Identificação, caracterização e controle in vitro de Phoma pedeiae (Aveskamp, Gruyter & Verkley) associado à Cedrela fissilis Vell.
author Dutra, Adriana Falcão
author_facet Dutra, Adriana Falcão
author_role author
dc.contributor.none.fl_str_mv Muniz, Marlove Fatima Brião
http://lattes.cnpq.br/3148312031889388
Oliveira, Andreia Mara Rotta de
http://lattes.cnpq.br/4046790504087569
Blume, Elena
http://lattes.cnpq.br/3299996746493028
Steffen, Gerusa Pauli Kist
http://lattes.cnpq.br/6467625663365353
Kulczynski, Stela Maris
http://lattes.cnpq.br/0604511258822899
dc.contributor.author.fl_str_mv Dutra, Adriana Falcão
dc.subject.por.fl_str_mv Fitopatologia
Controle de doenças
Óleos essenciais
Phytopathology
Disease control
Essencial oils
CNPQ::CIENCIAS AGRARIAS::RECURSOS FLORESTAIS E ENGENHARIA FLORESTAL
topic Fitopatologia
Controle de doenças
Óleos essenciais
Phytopathology
Disease control
Essencial oils
CNPQ::CIENCIAS AGRARIAS::RECURSOS FLORESTAIS E ENGENHARIA FLORESTAL
description The species Cedrela fissilis Vell. (cedro) presents good growth, producing wood of excellent quality, causing intense exploitation by the loggers. One of the main problems that compromise the production of wood is the occurrence of pests and diseases, causing serious damages to the host and changes in the physiological processes, leading to the death of the plant. Thus, the study of the biological activity of essential oils (OEs) of plants can constitute a potential form of control of diseases in plants. The objective of this study was the identification and morphological and molecular characterization of Phoma spp. associated with Cedrela fissilis Vell., and to evaluate the antifungal activity of cedro OE in the in vitro control of Phoma pedeiae. Leaflets were collected from adult trees with symptoms of the disease (Santa Maria, RS) and sent to the Elocy Minussi Phytopathology Laboratory (UFSM) for isolation of the fungus. The material was observed on stereoscopic and optical microscopes, and when fungal structures were present, they were transferred to Petri dishes containing BDA. After 14 days of development, the fungus was purified by monosporic culture and transferred to the BDA, CA and V8 media for morphological and molecular characterization. The pathogenicity of the fungus was performed by sprinkling on leaves for two isolates. The morphological characteristics analyzed were: mycelial growth (MG), mycelial growth rate (MGR) and growth rate index (GRI), sporulation, colony pigmentation and conidia characterization. OE extraction was carried out in the Laboratory of Extractive Plants, and OE was extracted from young cedro leaflets, in the spring season of 2015, 2016 and 2017, by hydrodistillation. The in vitro antifungal activity of leaflets OE was on pure colonies of Phoma pedeiae, at concentrations of 0.5, 0.75, 1.0 and 1.5 μL mL-1, in addition to positive and negative controls. Daily measurements of the colony diameter were carried out for 14 days, with MG, MGR, GRI and inhibition of mycelial growth. Sequences were deposited in GenBank and 70 and 81% similarly, with the fungus Didymella pedeiae, sexual form of Phoma pedeiae, by means of the sequencing of genes from the ITS and β-tubulin regions. The symptoms caused by the fungus started at 20 days after inoculation, not differing between the two isolates, causing the death of the individuals in 50 days. The fungus presented rounded pycnidia of black coloration, white mycelium and oblong oval conidia, without septum and hyaline. Sporulation in the V8 culture medium was superior to the BDA and CA medium. At the end of the three-year evaluation, 45 OE components of cedar leaflets were identified, corresponding to approximately 90% of the total OE composition, with the predominance of β-caryophyllene, spathulenol and caryophyllene oxide. The cedro OE presented relative antifungal activity against P. pedeiae starting at the concentration of 0.5 μL mL-1. There fore, it can be indicated for use in the control of pathogens, because it is a secondary metabolite, not causing damage to the environment.
publishDate 2018
dc.date.none.fl_str_mv 2018-12-14T17:05:39Z
2018-12-14T17:05:39Z
2018-02-26
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 http://repositorio.ufsm.br/handle/1/15107
url http://repositorio.ufsm.br/handle/1/15107
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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