Caracterização morfológica, molecular e patogênica de isolados de colletotrichum spp. de mangueira e a atividade inibitória de óleos essenciais sobre este patógeno

Detalhes bibliográficos
Autor(a) principal: Silva Neto, Jorge Alves
Data de Publicação: 2020
Tipo de documento: Dissertação
Idioma: por
Título da fonte: Repositório Digital da Universidade Federal Rural do Semi-Árido (RDU)
Texto Completo: https://repositorio.ufersa.edu.br/handle/prefix/5546
Resumo: The mango tree (Mangifera indica L.) stands out as one of the most important tropical fruit trees in the world. Anthracnose, caused by phytopathogenic fungi of the genus Colletotrichum, is a disease mainly related to post-harvest periods, directly affecting this crop. Therefore, this study aimed at the morphological, molecular and pathogenic characterization of Colletotrichum spp. of hoses and the inhibitory activity of essential oils on this pathogen. Fungal discs of 5 mm were placed in Petri dishes to evaluate the growth speed and colony color. Microcultures were carried out to analyze the shapes and dimensions of conidia and appressoria. For molecular analysis, the isolates were previously identified with primers specific gene of gender and species for C. gloeosporioides. Nine ISSR markers (Inter Simple Sequence Repeats) and 12 RAPD markers (Random Amplified Polymorphic DNA) were used to identify the genetic diversity of the 28 isolates. In the morphological evaluation, from the found structures, colony color, conidia, appressoria and growth rate, 28 isolates were identified belonging to the species C. gloeosporioides and later confirmed with specific primers genes. The RAPD and ISSR markers were considered satisfactory in detecting genetic diversity. The combination of the markers generated a variation of similarity from 0.23 (between isolates 12 and 21) to 0.71 (between isolates 2 and 4), thereby identifying high level of variability among fungi, where the dissimilarity was higher to 75%. The generated dendogram revealed the formation of 8 main groups. There were no genetic clusters correlated with the collection site, thus demonstrating the great genetic diversity even among individuals from the same location. The pathogenicity of C. gloeosporioides isolates was evaluated in Tommy Atkins mango fruits by introducing 7 mm mycelial discs. For the in vitro test, three more pathogenic isolates were used to assess inhibition with the oils of ginger (Zingiber officinale), dragon blood (Croton Lenchleri) and the combination of both at concentrations of 0.2; 0.4; 0.6 and 0.8%. The same treatments were used for the in vivo test. After the introduction of mycelial discs in mango fruits, the different concentrations of oils were spread on the inoculated surface and maintained during seven days after the evaluation. Based on the pathogenicity test, most isolates showed low virulence in the cultivar Tommy Atkins. However, some isolates have been shown to be highly pathogenic to this cultivar. In in vitro tests, there were significant differences between treatments, with the highest mean inhibition being 28.5% found in the combination of 0.3% ginger plus 0.3% dragon blood. For in vivo tests, all treatments were highly effective in Tommy mango, with 100% inhibition of fungal growth in the fruit. The data generated in this work allow us to conclude that there is variation not only genetic, but also in the level of pathogenicity among isolates of C. gloeosporioides collected in commercial mango plantations and that not always single control measures can be sufficient or 100% effective in the fight this disease. The inhibiting effect of ginger oil and dragon blood on the evaluated isolates reveals that these can be an effective method of controlling anthracnose
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spelling Caracterização morfológica, molecular e patogênica de isolados de colletotrichum spp. de mangueira e a atividade inibitória de óleos essenciais sobre este patógenoFitopatógenosDiversidade genéticaPatogenicidadePhytopathogensGenetical diversityPathogeniCNPQ::CIENCIAS AGRARIAS::AGRONOMIAThe mango tree (Mangifera indica L.) stands out as one of the most important tropical fruit trees in the world. Anthracnose, caused by phytopathogenic fungi of the genus Colletotrichum, is a disease mainly related to post-harvest periods, directly affecting this crop. Therefore, this study aimed at the morphological, molecular and pathogenic characterization of Colletotrichum spp. of hoses and the inhibitory activity of essential oils on this pathogen. Fungal discs of 5 mm were placed in Petri dishes to evaluate the growth speed and colony color. Microcultures were carried out to analyze the shapes and dimensions of conidia and appressoria. For molecular analysis, the isolates were previously identified with primers specific gene of gender and species for C. gloeosporioides. Nine ISSR markers (Inter Simple Sequence Repeats) and 12 RAPD markers (Random Amplified Polymorphic DNA) were used to identify the genetic diversity of the 28 isolates. In the morphological evaluation, from the found structures, colony color, conidia, appressoria and growth rate, 28 isolates were identified belonging to the species C. gloeosporioides and later confirmed with specific primers genes. The RAPD and ISSR markers were considered satisfactory in detecting genetic diversity. The combination of the markers generated a variation of similarity from 0.23 (between isolates 12 and 21) to 0.71 (between isolates 2 and 4), thereby identifying high level of variability among fungi, where the dissimilarity was higher to 75%. The generated dendogram revealed the formation of 8 main groups. There were no genetic clusters correlated with the collection site, thus demonstrating the great genetic diversity even among individuals from the same location. The pathogenicity of C. gloeosporioides isolates was evaluated in Tommy Atkins mango fruits by introducing 7 mm mycelial discs. For the in vitro test, three more pathogenic isolates were used to assess inhibition with the oils of ginger (Zingiber officinale), dragon blood (Croton Lenchleri) and the combination of both at concentrations of 0.2; 0.4; 0.6 and 0.8%. The same treatments were used for the in vivo test. After the introduction of mycelial discs in mango fruits, the different concentrations of oils were spread on the inoculated surface and maintained during seven days after the evaluation. Based on the pathogenicity test, most isolates showed low virulence in the cultivar Tommy Atkins. However, some isolates have been shown to be highly pathogenic to this cultivar. In in vitro tests, there were significant differences between treatments, with the highest mean inhibition being 28.5% found in the combination of 0.3% ginger plus 0.3% dragon blood. For in vivo tests, all treatments were highly effective in Tommy mango, with 100% inhibition of fungal growth in the fruit. The data generated in this work allow us to conclude that there is variation not only genetic, but also in the level of pathogenicity among isolates of C. gloeosporioides collected in commercial mango plantations and that not always single control measures can be sufficient or 100% effective in the fight this disease. The inhibiting effect of ginger oil and dragon blood on the evaluated isolates reveals that these can be an effective method of controlling anthracnoseA mangueira (Mangifera indica L.) se destaca como uma das fruteiras tropicais de maior importância no mundo. A antracnose, causada pelos fungos fitopatogênicos do gênero Colletotrichum, é uma doença relacionada principalmente a períodos de pós-colheita, afetando diretamente esta cultura. Diante disso, esse estudo objetivou a caracterização morfológica, molecular e patogênica de isolados de Colletotrichum spp. de mangueiras e a atividade inibitória de óleos essenciais sobre esse patógeno. Discos fúngicos de 5 mm foram dispostos em placas de Petri para avaliação da velocidade de crescimento e coloração das colônias. Foram realizados microcultivos para análises dos formatos e dimensões de conídios e apressórios. Para as análises moleculares, os isolados foram previamente identificados com primers gene específico de gênero e espécie para C. gloeosporioides. Nove iniciadores ISSR (Inter Simple Sequence Repeats) e 12 iniciadores RAPD (Random Amplified Polymorphic DNA) foram utilizados para identificar a diversidade genética dos 28 isolados. Na avaliação morfológica, a partir das estruturas encontradas, coloração das colônias, conídios, apressórios e velocidade de crescimento, 28 isolados foram identificados como pertencentes à espécie C. gloeosporioides, sendo posteriormente confirmados com os primers genes específicos. Os iniciadores RAPD e ISSR foram considerados satisfatórios em identificar a diversidade genética. A combinação dos iniciadores gerou variação de similaridade de 0,23 (entre os isolados 12 e 21) a 0,71 (entre os isolados 2 e 4), identificando, com isso, alto nível de variabilidade entre os fungos, onde a dissimilaridade foi superior a 75%. O dendograma gerado revelou a formação de oito grupos principais. Não foram observados agrupamentos genéticos correlacionados com o local da coleta, mostrando, assim, a grande diversidade genética mesmo entre indivíduos de mesma localidade. A patogenicidade de isolados de C. gloeosporioides foi avaliada em frutas de manga Tommy Atkins mediante introdução de discos miceliais de 7 mm. Para o teste in vitro, três isolados mais patogênicos foram utilizados para a avaliação de inibição com os óleos de gengibre (Zingiber officinale), sangue de dragão (Croton Lenchleri) e a combinação de ambos nas concentrações de 0,2; 0,4; 0,6 e 0,8%. Os mesmos tratamentos foram utilizados para o teste in vivo. Após a introdução de discos miceliais em frutos de manga, as diferentes concentrações de óleos foram espalhadas na superfície inoculada e mantidas por sete dias após a avaliação. Com base no teste de patogenicidade, a maioria dos isolados apresentou baixa virulência na cultivar Tommy Atkins. Entretanto, alguns isolados se mostraram altamente patogênicos a essa cultivar. Nos testes in vitro, houve diferenças significativas entre os tratamentos, sendo que a maior inibição média foi de 28,5%, encontrada na combinação de 0,3% de gengibre mais 0,3% de sangue de dragão. Para os testes in vivo, todos os tratamentos se mostraram altamente eficazes em manga Tommy, apresentando 100% de inibição do crescimento fúngico no fruto. Os dados gerados neste trabalho permitem concluir que há variação não apenas genética, como também no nível de patogenicidade entre os isolados de C. gloeosporioides coletados em plantios comerciais de manga e que nem sempre medidas únicas de controle podem ser suficientes ou 100% eficazes no combate a essa enfermidade. O efeito de inibição dos óleos de gengibre e sangue de dragão nos isolados avaliados revela que este pode ser um eficaz método de controle da antracnoseCoordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPESUniversidade Federal Rural do Semi-ÁridoBrasilCentro de Ciências Agrárias - CCAUFERSAPrograma de Pós-Graduação em FitotecniaAmbrósio, Marcia Michelle de Queiroz96726539487http://lattes.cnpq.br/5986626995999424Holanda, Ioná Santos Araújo65180062500http://lattes.cnpq.br/2079609001869001Ambrósio, Marcia Michelle de Queiroz96726539487http://lattes.cnpq.br/5986626995999424Negreiros, Andréia Mitsa Paiva08897954448http://lattes.cnpq.br/3271764526148277Walter, Rafael04318096157http://lattes.cnpq.br/0927184479047358Silva Neto, Jorge Alves2020-09-30T10:41:59Z2020-09-292020-09-30T10:41:59Z2020-03-11info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisapplication/pdfSilva Neto (2020) (SILVA NETO, 2020)https://repositorio.ufersa.edu.br/handle/prefix/5546porSILVA NETO, Jorge Alves da. Caracterização morfológica, molecular e patogênica de isolados de colletotrichum spp. de mangueira e a atividade inibitória de óleos essenciais sobre este patógeno. 2020. 58 f. Dissertação (Mestrado em Fitotecnia), Universidade Federal Rural do Semi-Árido, Mossoró, 2020.CC-BY-SAinfo:eu-repo/semantics/openAccessreponame:Repositório Digital da Universidade Federal Rural do Semi-Árido (RDU)instname:Universidade Federal Rural do Semi-Árido (UFERSA)instacron:UFERSA2023-10-30T20:27:22Zoai:repositorio.ufersa.edu.br:prefix/5546Repositório Institucionalhttps://repositorio.ufersa.edu.br/PUBhttps://repositorio.ufersa.edu.br/server/oai/requestrepositorio@ufersa.edu.br || admrepositorio@ufersa.edu.bropendoar:2023-10-30T20:27:22Repositório Digital da Universidade Federal Rural do Semi-Árido (RDU) - Universidade Federal Rural do Semi-Árido (UFERSA)false
dc.title.none.fl_str_mv Caracterização morfológica, molecular e patogênica de isolados de colletotrichum spp. de mangueira e a atividade inibitória de óleos essenciais sobre este patógeno
title Caracterização morfológica, molecular e patogênica de isolados de colletotrichum spp. de mangueira e a atividade inibitória de óleos essenciais sobre este patógeno
spellingShingle Caracterização morfológica, molecular e patogênica de isolados de colletotrichum spp. de mangueira e a atividade inibitória de óleos essenciais sobre este patógeno
Silva Neto, Jorge Alves
Fitopatógenos
Diversidade genética
Patogenicidade
Phytopathogens
Genetical diversity
Pathogeni
CNPQ::CIENCIAS AGRARIAS::AGRONOMIA
title_short Caracterização morfológica, molecular e patogênica de isolados de colletotrichum spp. de mangueira e a atividade inibitória de óleos essenciais sobre este patógeno
title_full Caracterização morfológica, molecular e patogênica de isolados de colletotrichum spp. de mangueira e a atividade inibitória de óleos essenciais sobre este patógeno
title_fullStr Caracterização morfológica, molecular e patogênica de isolados de colletotrichum spp. de mangueira e a atividade inibitória de óleos essenciais sobre este patógeno
title_full_unstemmed Caracterização morfológica, molecular e patogênica de isolados de colletotrichum spp. de mangueira e a atividade inibitória de óleos essenciais sobre este patógeno
title_sort Caracterização morfológica, molecular e patogênica de isolados de colletotrichum spp. de mangueira e a atividade inibitória de óleos essenciais sobre este patógeno
author Silva Neto, Jorge Alves
author_facet Silva Neto, Jorge Alves
author_role author
dc.contributor.none.fl_str_mv Ambrósio, Marcia Michelle de Queiroz
96726539487
http://lattes.cnpq.br/5986626995999424
Holanda, Ioná Santos Araújo
65180062500
http://lattes.cnpq.br/2079609001869001
Ambrósio, Marcia Michelle de Queiroz
96726539487
http://lattes.cnpq.br/5986626995999424
Negreiros, Andréia Mitsa Paiva
08897954448
http://lattes.cnpq.br/3271764526148277
Walter, Rafael
04318096157
http://lattes.cnpq.br/0927184479047358
dc.contributor.author.fl_str_mv Silva Neto, Jorge Alves
dc.subject.por.fl_str_mv Fitopatógenos
Diversidade genética
Patogenicidade
Phytopathogens
Genetical diversity
Pathogeni
CNPQ::CIENCIAS AGRARIAS::AGRONOMIA
topic Fitopatógenos
Diversidade genética
Patogenicidade
Phytopathogens
Genetical diversity
Pathogeni
CNPQ::CIENCIAS AGRARIAS::AGRONOMIA
description The mango tree (Mangifera indica L.) stands out as one of the most important tropical fruit trees in the world. Anthracnose, caused by phytopathogenic fungi of the genus Colletotrichum, is a disease mainly related to post-harvest periods, directly affecting this crop. Therefore, this study aimed at the morphological, molecular and pathogenic characterization of Colletotrichum spp. of hoses and the inhibitory activity of essential oils on this pathogen. Fungal discs of 5 mm were placed in Petri dishes to evaluate the growth speed and colony color. Microcultures were carried out to analyze the shapes and dimensions of conidia and appressoria. For molecular analysis, the isolates were previously identified with primers specific gene of gender and species for C. gloeosporioides. Nine ISSR markers (Inter Simple Sequence Repeats) and 12 RAPD markers (Random Amplified Polymorphic DNA) were used to identify the genetic diversity of the 28 isolates. In the morphological evaluation, from the found structures, colony color, conidia, appressoria and growth rate, 28 isolates were identified belonging to the species C. gloeosporioides and later confirmed with specific primers genes. The RAPD and ISSR markers were considered satisfactory in detecting genetic diversity. The combination of the markers generated a variation of similarity from 0.23 (between isolates 12 and 21) to 0.71 (between isolates 2 and 4), thereby identifying high level of variability among fungi, where the dissimilarity was higher to 75%. The generated dendogram revealed the formation of 8 main groups. There were no genetic clusters correlated with the collection site, thus demonstrating the great genetic diversity even among individuals from the same location. The pathogenicity of C. gloeosporioides isolates was evaluated in Tommy Atkins mango fruits by introducing 7 mm mycelial discs. For the in vitro test, three more pathogenic isolates were used to assess inhibition with the oils of ginger (Zingiber officinale), dragon blood (Croton Lenchleri) and the combination of both at concentrations of 0.2; 0.4; 0.6 and 0.8%. The same treatments were used for the in vivo test. After the introduction of mycelial discs in mango fruits, the different concentrations of oils were spread on the inoculated surface and maintained during seven days after the evaluation. Based on the pathogenicity test, most isolates showed low virulence in the cultivar Tommy Atkins. However, some isolates have been shown to be highly pathogenic to this cultivar. In in vitro tests, there were significant differences between treatments, with the highest mean inhibition being 28.5% found in the combination of 0.3% ginger plus 0.3% dragon blood. For in vivo tests, all treatments were highly effective in Tommy mango, with 100% inhibition of fungal growth in the fruit. The data generated in this work allow us to conclude that there is variation not only genetic, but also in the level of pathogenicity among isolates of C. gloeosporioides collected in commercial mango plantations and that not always single control measures can be sufficient or 100% effective in the fight this disease. The inhibiting effect of ginger oil and dragon blood on the evaluated isolates reveals that these can be an effective method of controlling anthracnose
publishDate 2020
dc.date.none.fl_str_mv 2020-09-30T10:41:59Z
2020-09-29
2020-09-30T10:41:59Z
2020-03-11
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 Silva Neto (2020) (SILVA NETO, 2020)
https://repositorio.ufersa.edu.br/handle/prefix/5546
identifier_str_mv Silva Neto (2020) (SILVA NETO, 2020)
url https://repositorio.ufersa.edu.br/handle/prefix/5546
dc.language.iso.fl_str_mv por
language por
dc.relation.none.fl_str_mv SILVA NETO, Jorge Alves da. Caracterização morfológica, molecular e patogênica de isolados de colletotrichum spp. de mangueira e a atividade inibitória de óleos essenciais sobre este patógeno. 2020. 58 f. Dissertação (Mestrado em Fitotecnia), Universidade Federal Rural do Semi-Árido, Mossoró, 2020.
dc.rights.driver.fl_str_mv CC-BY-SA
info:eu-repo/semantics/openAccess
rights_invalid_str_mv CC-BY-SA
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Universidade Federal Rural do Semi-Árido
Brasil
Centro de Ciências Agrárias - CCA
UFERSA
Programa de Pós-Graduação em Fitotecnia
publisher.none.fl_str_mv Universidade Federal Rural do Semi-Árido
Brasil
Centro de Ciências Agrárias - CCA
UFERSA
Programa de Pós-Graduação em Fitotecnia
dc.source.none.fl_str_mv reponame:Repositório Digital da Universidade Federal Rural do Semi-Árido (RDU)
instname:Universidade Federal Rural do Semi-Árido (UFERSA)
instacron:UFERSA
instname_str Universidade Federal Rural do Semi-Árido (UFERSA)
instacron_str UFERSA
institution UFERSA
reponame_str Repositório Digital da Universidade Federal Rural do Semi-Árido (RDU)
collection Repositório Digital da Universidade Federal Rural do Semi-Árido (RDU)
repository.name.fl_str_mv Repositório Digital da Universidade Federal Rural do Semi-Árido (RDU) - Universidade Federal Rural do Semi-Árido (UFERSA)
repository.mail.fl_str_mv repositorio@ufersa.edu.br || admrepositorio@ufersa.edu.br
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