Loquat (Eriobotrya Japonica) microbiota : screening of potential biocontrol agents against Erwinia amylovora

Detalhes bibliográficos
Autor(a) principal: Azevedo, Telma Maria Sampaio
Data de Publicação: 2017
Tipo de documento: Dissertação
Idioma: por
Título da fonte: Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
Texto Completo: http://hdl.handle.net/20.500.11960/1869
Resumo: Fire blight, caused by the bacterium Erwinia amylovora, is one of the most destructive diseases that affect crops of the Rosaceae family, representing a serious threat to orchards and ornamental species of a great economic interest. Due to its pathogenicity, E. amylovora is considered a quarantine organism in the European Union. Portugal is, since 2008, recognized as a “Protected Area” for this bacterium. The lack of available efficient phytochemicals and antibiotics, as well as their high toxicity and resistance, have led to the innumerous research work to screen for new organisms with a biological control action. Nowadays biological control active ingredients are already present in some commercialized products, used as a preventive measure against E. amylovora. However, further studies are necessary in investigating new strains or species with biopesticide action for specific crops. The main objective of this work was to select for possible antagonists against E. amylovora from microbiota obtained from loquat (Eriobotrya japonica) flowers and leaves. For this reason, distinct isolation methodologies, characterization and ex vivo assays were performed in fruits and flowers in order to evaluate the efficacy of the strains in study on delaying the onset of symptoms caused by pathogen infection, as well as the incidence and severity of this infection. Out of the 173 bacterial isolates studied, 4 species (Rosenbergiella epipactidis, Pseudomonas rhizosphaerae, Curtobacterium flaccumfaciens and Enterobacter cancerogenus) were selected since they delayed the onset of fruit necrosis, being associated with low levels of incidence and severity of disease infection (high efficacy of antagonistic activity), as well as no pathogenicity. Since flower stigmas are the more appropriate organs to E. amylovora colonization and development, two flower bioassays were conducted to assess the antagonistic activity of these bacterial species: in complete flowers (R. epipactidis, P. rhizosphaerae, C. flaccumfaciens and E. cancerogenus) and in flowers only with hypanthium and peduncle (R. epipactidis and P. rhizosphaerae). The molecular analysis by real-time PCR to quantify E. amylovora concentrations after treating the flowers with a selected bacterial antagonistic species did not show a relationship between the E. amylovora concentration and the observed symptoms in both flower bioassays.
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spelling Loquat (Eriobotrya Japonica) microbiota : screening of potential biocontrol agents against Erwinia amylovoraErwinia amylovoraMicrobiotaFire blightPathogenBactériaEnterobacter cancerogenusCurtobacterium flaccumfaciensRosenbergiella epipactidisPseudomonasRhizosphaeraeFire blight, caused by the bacterium Erwinia amylovora, is one of the most destructive diseases that affect crops of the Rosaceae family, representing a serious threat to orchards and ornamental species of a great economic interest. Due to its pathogenicity, E. amylovora is considered a quarantine organism in the European Union. Portugal is, since 2008, recognized as a “Protected Area” for this bacterium. The lack of available efficient phytochemicals and antibiotics, as well as their high toxicity and resistance, have led to the innumerous research work to screen for new organisms with a biological control action. Nowadays biological control active ingredients are already present in some commercialized products, used as a preventive measure against E. amylovora. However, further studies are necessary in investigating new strains or species with biopesticide action for specific crops. The main objective of this work was to select for possible antagonists against E. amylovora from microbiota obtained from loquat (Eriobotrya japonica) flowers and leaves. For this reason, distinct isolation methodologies, characterization and ex vivo assays were performed in fruits and flowers in order to evaluate the efficacy of the strains in study on delaying the onset of symptoms caused by pathogen infection, as well as the incidence and severity of this infection. Out of the 173 bacterial isolates studied, 4 species (Rosenbergiella epipactidis, Pseudomonas rhizosphaerae, Curtobacterium flaccumfaciens and Enterobacter cancerogenus) were selected since they delayed the onset of fruit necrosis, being associated with low levels of incidence and severity of disease infection (high efficacy of antagonistic activity), as well as no pathogenicity. Since flower stigmas are the more appropriate organs to E. amylovora colonization and development, two flower bioassays were conducted to assess the antagonistic activity of these bacterial species: in complete flowers (R. epipactidis, P. rhizosphaerae, C. flaccumfaciens and E. cancerogenus) and in flowers only with hypanthium and peduncle (R. epipactidis and P. rhizosphaerae). The molecular analysis by real-time PCR to quantify E. amylovora concentrations after treating the flowers with a selected bacterial antagonistic species did not show a relationship between the E. amylovora concentration and the observed symptoms in both flower bioassays.O fogo bacteriano, causado pela bactéria Erwinia amylovora, é uma das doenças mais destrutivas de inúmeras culturas da família das Rosáceas, representando uma séria ameaça a árvores de fruto e espécies ornamentais de grande interesse económico. Devido à sua elevada patogenicidade, E. amylovora é considerada um organismo de quarentena na União Europeia. Portugal é, desde 2008, reconhecido como “Zona Protegida” para a bactéria. Os resultados pouco satisfatórios obtidos pela utilização de fitoquímicos e antibióticos, acrescidos da elevada toxicidade e resistências dos mesmos no combate à bactéria, conduziram a diversas pesquisas por novos organismos com modo de ação biológica. Atualmente já são comercializados alguns produtos que possuem como substâncias ativas agentes de controlo biológico, usados como ação preventiva contra E. amylovora. Contudo, são necessários mais estudos na procura de novas estirpes ou espécies com ação biopesticida. Este trabalho teve como principal objetivo selecionar possíveis antagonistas contra E. amylovora a partir da microbiota de flores e folhas de magnórios (Eriobotrya japonica). Para isso, foram realizadas diversas metodologias de isolamento, caracterização e ensaios ex vivo em frutos e flores para avaliação da eficácia das estirpes em estudo na redução do aparecimento de sintomas resultantes da infeção pelo patogéneo, bem como incidência e severidade desta infeção. Em 173 isolados bacterianos estudados, 4 espécies (Rosenbergiella epipactidis, Pseudomonas rhizosphaerae, Curtobacterium flaccumfaciens e Enterobacter cancerogenus) foram selecionadas uma vez que atrasaram mais o aparecimento de necrose em fruto, tendo conduzido a níveis de incidência e severidade baixos da doença (elevada eficácia de atividade antagonista), além de não apresentarem patogenicidade. Sendo os estigmas das flores os órgãos mais propícios para a colonização e desenvolvimento de E. amylovora, procedeu-se à realização de dois bioensaios em flores para testar a ação antagonista destas espécies: em flores completas (R. epipactidis, P. rhizosphaerae, C. flaccumfaciens e E. cancerogenus) e em flores apenas com o hipanto e pedúnculo (R. epipactidis, P. rhizosphaerae). A análise molecular através de PCR em tempo real, para quantificar a concentração de E. amylovora, após inoculação de flores com as espécies antagonistas selecionadas não evidenciou uma relação entre a concentração de E. amylovora e os sintomas observados em ambos os bioensaios realizados em flores.2017-06-20T15:12:10Z2017-02-17T00:00:00Z2017-02-17info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisapplication/pdfhttp://hdl.handle.net/20.500.11960/1869TID:201701898porAzevedo, Telma Maria Sampaioinfo:eu-repo/semantics/openAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2023-03-21T14:36:18Zoai:repositorio.ipvc.pt:20.500.11960/1869Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-19T17:43:30.262712Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãofalse
dc.title.none.fl_str_mv Loquat (Eriobotrya Japonica) microbiota : screening of potential biocontrol agents against Erwinia amylovora
title Loquat (Eriobotrya Japonica) microbiota : screening of potential biocontrol agents against Erwinia amylovora
spellingShingle Loquat (Eriobotrya Japonica) microbiota : screening of potential biocontrol agents against Erwinia amylovora
Azevedo, Telma Maria Sampaio
Erwinia amylovora
Microbiota
Fire blight
Pathogen
Bactéria
Enterobacter cancerogenus
Curtobacterium flaccumfaciens
Rosenbergiella epipactidis
Pseudomonas
Rhizosphaerae
title_short Loquat (Eriobotrya Japonica) microbiota : screening of potential biocontrol agents against Erwinia amylovora
title_full Loquat (Eriobotrya Japonica) microbiota : screening of potential biocontrol agents against Erwinia amylovora
title_fullStr Loquat (Eriobotrya Japonica) microbiota : screening of potential biocontrol agents against Erwinia amylovora
title_full_unstemmed Loquat (Eriobotrya Japonica) microbiota : screening of potential biocontrol agents against Erwinia amylovora
title_sort Loquat (Eriobotrya Japonica) microbiota : screening of potential biocontrol agents against Erwinia amylovora
author Azevedo, Telma Maria Sampaio
author_facet Azevedo, Telma Maria Sampaio
author_role author
dc.contributor.author.fl_str_mv Azevedo, Telma Maria Sampaio
dc.subject.por.fl_str_mv Erwinia amylovora
Microbiota
Fire blight
Pathogen
Bactéria
Enterobacter cancerogenus
Curtobacterium flaccumfaciens
Rosenbergiella epipactidis
Pseudomonas
Rhizosphaerae
topic Erwinia amylovora
Microbiota
Fire blight
Pathogen
Bactéria
Enterobacter cancerogenus
Curtobacterium flaccumfaciens
Rosenbergiella epipactidis
Pseudomonas
Rhizosphaerae
description Fire blight, caused by the bacterium Erwinia amylovora, is one of the most destructive diseases that affect crops of the Rosaceae family, representing a serious threat to orchards and ornamental species of a great economic interest. Due to its pathogenicity, E. amylovora is considered a quarantine organism in the European Union. Portugal is, since 2008, recognized as a “Protected Area” for this bacterium. The lack of available efficient phytochemicals and antibiotics, as well as their high toxicity and resistance, have led to the innumerous research work to screen for new organisms with a biological control action. Nowadays biological control active ingredients are already present in some commercialized products, used as a preventive measure against E. amylovora. However, further studies are necessary in investigating new strains or species with biopesticide action for specific crops. The main objective of this work was to select for possible antagonists against E. amylovora from microbiota obtained from loquat (Eriobotrya japonica) flowers and leaves. For this reason, distinct isolation methodologies, characterization and ex vivo assays were performed in fruits and flowers in order to evaluate the efficacy of the strains in study on delaying the onset of symptoms caused by pathogen infection, as well as the incidence and severity of this infection. Out of the 173 bacterial isolates studied, 4 species (Rosenbergiella epipactidis, Pseudomonas rhizosphaerae, Curtobacterium flaccumfaciens and Enterobacter cancerogenus) were selected since they delayed the onset of fruit necrosis, being associated with low levels of incidence and severity of disease infection (high efficacy of antagonistic activity), as well as no pathogenicity. Since flower stigmas are the more appropriate organs to E. amylovora colonization and development, two flower bioassays were conducted to assess the antagonistic activity of these bacterial species: in complete flowers (R. epipactidis, P. rhizosphaerae, C. flaccumfaciens and E. cancerogenus) and in flowers only with hypanthium and peduncle (R. epipactidis and P. rhizosphaerae). The molecular analysis by real-time PCR to quantify E. amylovora concentrations after treating the flowers with a selected bacterial antagonistic species did not show a relationship between the E. amylovora concentration and the observed symptoms in both flower bioassays.
publishDate 2017
dc.date.none.fl_str_mv 2017-06-20T15:12:10Z
2017-02-17T00:00:00Z
2017-02-17
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