Análise do envolvimento de proteínas do látex de Calotropis procera na defesa vegetal face a estresses biótico e abiótico

Detalhes bibliográficos
Ano de defesa: 2019
Autor(a) principal: Souza, Isabel Cristina da Costa
Orientador(a): Ramos, Márcio Viana
Banca de defesa: Não Informado pela instituição
Tipo de documento: Tese
Tipo de acesso: Acesso aberto
Idioma: por
Instituição de defesa: Não Informado pela instituição
Programa de Pós-Graduação: Não Informado pela instituição
Departamento: Não Informado pela instituição
País: Não Informado pela instituição
Palavras-chave em Português:
Link de acesso: http://www.repositorio.ufc.br/handle/riufc/46145
Resumo: ABSTRACT Throughout evolution, the plants have been developing defense mechanisms for their protection, an example of this is the production of latex as a vegetal resource in the defense against herbivores and phytopathogens. The chemical and functional diversity of the laticiferous fluids is being studied, with emphasis on secondary metabolites and defense proteins. The species Calotropis procera belongs to the family Apocynaceae and is prominent in latex production. Three classes of proteins isolated from latex: proteases, osmotin and chitinases. However, the objective of this study was to trace a relationship between the proteins present in the latex of C. procera and their role in plant defense against biotic and abiotic stresses. For this, methodologies for the in vitro control of the mycelial growth and germination of the phytopathogenic fungi Colletotrichum gloeosporioides, Fusarium tupiense e F. oxysporum f. sp. cubense testing the synergistic effect of total proteins and their isolated fractions. Thus, it was verified the total proteins present a greater effect when compared to the latex fractions of C. procera, but the combination of the proteases with the osmotin enhaces the effect compared to the isolated osmotin. The total proteins of C. procera latex were tested as elicitors in the defense of banana seedlings challenged with the fungus F. oxysporum f. sp. cubense. It was observed that plants who were previously treated with latex proteins showed a reduction in the occurrence and severity of the disease, maintenance of growth, reduction in membrane damage and increase in the activity of the antioxidant system enzymes. Osmotins are defense-related proteins when it comes to biotic and abiotic stresses. To test this hypothesis to the sequence of the process of Nicotiana tabacum with the osmotin of C. procera (CpOsm). After confirming the transformation of the first generation of plants, the total proteins of tobacco leaves were extracted and tested in the inhibition of the germination and mycelial growth of the fungi F. tupienses and F. oxysporum f. sp. cubense. The results show plants transformed with the CpOsm gene inhibit the mycelial growth of phytopathogenic fungi with greater efficiency when compared to the untransformed plants. The transformed and control plants were submitted to salinity resistance test. For this, the plants were maintained in nutrient medium MS with 300 mM NaCl for seven days, after the transformed plants had no difference in growth and number of leaves, however they were able to increase the concentration of free proline when compared to the non-plants transformed. It is possible to measure a probable effect of C. procera latex proteins on the plant defense mechanism against the attack of phytopathogens and to relate the CpOsm protein with mechanisms of action of protection of plants to saline stress and attacks by phytopathogens.
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spelling Souza, Isabel Cristina da CostaRamos, Márcio Viana2019-09-26T14:17:48Z2019-09-26T14:17:48Z2019SOUZA, Isabel Cristina da Costa. Análise do envolvimento de proteínas do látex de Calotropis procera na defesa vegetal face a estresses biótico e abiótico. 2019. 119f. Tese (Doutorado em Bioquímica) - Universidade Federal do Ceará, Fortaleza, 2019http://www.repositorio.ufc.br/handle/riufc/46145ABSTRACT Throughout evolution, the plants have been developing defense mechanisms for their protection, an example of this is the production of latex as a vegetal resource in the defense against herbivores and phytopathogens. The chemical and functional diversity of the laticiferous fluids is being studied, with emphasis on secondary metabolites and defense proteins. The species Calotropis procera belongs to the family Apocynaceae and is prominent in latex production. Three classes of proteins isolated from latex: proteases, osmotin and chitinases. However, the objective of this study was to trace a relationship between the proteins present in the latex of C. procera and their role in plant defense against biotic and abiotic stresses. For this, methodologies for the in vitro control of the mycelial growth and germination of the phytopathogenic fungi Colletotrichum gloeosporioides, Fusarium tupiense e F. oxysporum f. sp. cubense testing the synergistic effect of total proteins and their isolated fractions. Thus, it was verified the total proteins present a greater effect when compared to the latex fractions of C. procera, but the combination of the proteases with the osmotin enhaces the effect compared to the isolated osmotin. The total proteins of C. procera latex were tested as elicitors in the defense of banana seedlings challenged with the fungus F. oxysporum f. sp. cubense. It was observed that plants who were previously treated with latex proteins showed a reduction in the occurrence and severity of the disease, maintenance of growth, reduction in membrane damage and increase in the activity of the antioxidant system enzymes. Osmotins are defense-related proteins when it comes to biotic and abiotic stresses. To test this hypothesis to the sequence of the process of Nicotiana tabacum with the osmotin of C. procera (CpOsm). After confirming the transformation of the first generation of plants, the total proteins of tobacco leaves were extracted and tested in the inhibition of the germination and mycelial growth of the fungi F. tupienses and F. oxysporum f. sp. cubense. The results show plants transformed with the CpOsm gene inhibit the mycelial growth of phytopathogenic fungi with greater efficiency when compared to the untransformed plants. The transformed and control plants were submitted to salinity resistance test. For this, the plants were maintained in nutrient medium MS with 300 mM NaCl for seven days, after the transformed plants had no difference in growth and number of leaves, however they were able to increase the concentration of free proline when compared to the non-plants transformed. It is possible to measure a probable effect of C. procera latex proteins on the plant defense mechanism against the attack of phytopathogens and to relate the CpOsm protein with mechanisms of action of protection of plants to saline stress and attacks by phytopathogens.Ao longo da evolução, as plantas desenvolveram diferentes estratégias de defesa para sua proteção. O látex é um composto presente em uma ampla variedade de espécies sendo um recurso para a defesa contra herbívoros e fitopatógenos. Sua diversidade química e funcional tem sido bastante estudada, contendo metabólitos secundários e proteínas predominam. A espécie Calotropis procera (Apocynaceae) é laticífera. Três classes de proteínas isoladas do látex dessa espécie já foram bem caracterizadas: proteases, osmotinas e quitinases. O objetivo desse estudo foi traçar uma correlação entre proteínas presentes no látex de C. procera e sua participação na defesa das plantas frente a estresses bióticos e abióticos. Foram realizados ensaios para mensurar o controle in vitro do crescimento micelial e germinação de esporos de fungos fitopatogênicos (Colletotrichum gloeosporioides, Fusarium tupiense e F. oxysporum f. sp. cubense) testando o efeito sinérgico das proteínas totais assim como suas frações isoladas. Dessa forma, foi verificado que as proteínas totais apresentam um maior efeito quando comparado com as proteínas fracionadas do látex. Porém a combinação das proteases com as osmotinas potencializam o efeito quando comparado com a ação isolada das osmotinas. As proteínas totais do látex foram testadas como agentes elicitores na defesa de mudas de bananeira desafiadas com o fungo F. oxysporum f. sp. cubense. Foi observado que as plantas que passaram por um tratamento prévio com proteínas do látex apresentaram uma redução na ocorrência e severidade da doença. Mantiveram o crescimento e redução no dano de membrana assim como aumento na atividade das enzimas do sistema antioxidante. Plantas de Nicotiana tabacum foram transformadas com o gene da osmotina do látex de C. procera (CpOsm). Após a confirmação da transformação da primeira geração de plantas, as proteínas totais de folhas de tabaco foram extraídas e testadas na inibição da germinação e crescimento micelial dos fungos F. tupiens e F. oxysporum f. sp. cubense. Os resultados mostram que as plantas transformadas com o gene CpOsm conseguiram inibir o crescimento micelial de fungos fitopatogênicos com maior eficiência quando comparadas com as plantas não transformadas. As plantas, transformadas e controle, foram submetidas a teste de resistência a salinidade. As plantas transformadas não exibiram diferença de tamanho e número de folhas, porém conseguiram aumentar a concentração de prolina livre quando comparada com as plantas não transformadas. É possível aferir um provável efeito das proteínas do látex de C. procera no mecanismo de defesa das plantas ao ataque de fitopatógenos e relacionar a proteína CpOsm com mecanismos de ação de proteção das plantas ao estresse salino e a ataques por fitopatógenos.fitopatógenosproteínas de defesatransformação genéticaAnálise do envolvimento de proteínas do látex de Calotropis procera na defesa vegetal face a estresses biótico e abióticoAnalysis of the involvement of latex proteins of Calotropis procera in vegetal defense face to biotic and abiotic stressesinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesisporreponame:Repositório Institucional da Universidade Federal do Ceará (UFC)instname:Universidade Federal do Ceará (UFC)instacron:UFCinfo:eu-repo/semantics/openAccessORIGINAL2019_tese_iccsouza.pdf2019_tese_iccsouza.pdfapplication/pdf3255018http://repositorio.ufc.br/bitstream/riufc/46145/7/2019_tese_iccsouza.pdf160ab2ffa652e9fd816ce494272fcee2MD57LICENSElicense.txtlicense.txttext/plain; charset=utf-81748http://repositorio.ufc.br/bitstream/riufc/46145/8/license.txt8a4605be74aa9ea9d79846c1fba20a33MD58riufc/461452019-09-26 11:17:48.957oai:repositorio.ufc.br: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Repositório InstitucionalPUBhttp://www.repositorio.ufc.br/ri-oai/requestbu@ufc.br || repositorio@ufc.bropendoar:2019-09-26T14:17:48Repositório Institucional da Universidade Federal do Ceará (UFC) - Universidade Federal do Ceará (UFC)false
dc.title.pt_BR.fl_str_mv Análise do envolvimento de proteínas do látex de Calotropis procera na defesa vegetal face a estresses biótico e abiótico
dc.title.en.pt_BR.fl_str_mv Analysis of the involvement of latex proteins of Calotropis procera in vegetal defense face to biotic and abiotic stresses
title Análise do envolvimento de proteínas do látex de Calotropis procera na defesa vegetal face a estresses biótico e abiótico
spellingShingle Análise do envolvimento de proteínas do látex de Calotropis procera na defesa vegetal face a estresses biótico e abiótico
Souza, Isabel Cristina da Costa
fitopatógenos
proteínas de defesa
transformação genética
title_short Análise do envolvimento de proteínas do látex de Calotropis procera na defesa vegetal face a estresses biótico e abiótico
title_full Análise do envolvimento de proteínas do látex de Calotropis procera na defesa vegetal face a estresses biótico e abiótico
title_fullStr Análise do envolvimento de proteínas do látex de Calotropis procera na defesa vegetal face a estresses biótico e abiótico
title_full_unstemmed Análise do envolvimento de proteínas do látex de Calotropis procera na defesa vegetal face a estresses biótico e abiótico
title_sort Análise do envolvimento de proteínas do látex de Calotropis procera na defesa vegetal face a estresses biótico e abiótico
author Souza, Isabel Cristina da Costa
author_facet Souza, Isabel Cristina da Costa
author_role author
dc.contributor.author.fl_str_mv Souza, Isabel Cristina da Costa
dc.contributor.advisor1.fl_str_mv Ramos, Márcio Viana
contributor_str_mv Ramos, Márcio Viana
dc.subject.por.fl_str_mv fitopatógenos
proteínas de defesa
transformação genética
topic fitopatógenos
proteínas de defesa
transformação genética
description ABSTRACT Throughout evolution, the plants have been developing defense mechanisms for their protection, an example of this is the production of latex as a vegetal resource in the defense against herbivores and phytopathogens. The chemical and functional diversity of the laticiferous fluids is being studied, with emphasis on secondary metabolites and defense proteins. The species Calotropis procera belongs to the family Apocynaceae and is prominent in latex production. Three classes of proteins isolated from latex: proteases, osmotin and chitinases. However, the objective of this study was to trace a relationship between the proteins present in the latex of C. procera and their role in plant defense against biotic and abiotic stresses. For this, methodologies for the in vitro control of the mycelial growth and germination of the phytopathogenic fungi Colletotrichum gloeosporioides, Fusarium tupiense e F. oxysporum f. sp. cubense testing the synergistic effect of total proteins and their isolated fractions. Thus, it was verified the total proteins present a greater effect when compared to the latex fractions of C. procera, but the combination of the proteases with the osmotin enhaces the effect compared to the isolated osmotin. The total proteins of C. procera latex were tested as elicitors in the defense of banana seedlings challenged with the fungus F. oxysporum f. sp. cubense. It was observed that plants who were previously treated with latex proteins showed a reduction in the occurrence and severity of the disease, maintenance of growth, reduction in membrane damage and increase in the activity of the antioxidant system enzymes. Osmotins are defense-related proteins when it comes to biotic and abiotic stresses. To test this hypothesis to the sequence of the process of Nicotiana tabacum with the osmotin of C. procera (CpOsm). After confirming the transformation of the first generation of plants, the total proteins of tobacco leaves were extracted and tested in the inhibition of the germination and mycelial growth of the fungi F. tupienses and F. oxysporum f. sp. cubense. The results show plants transformed with the CpOsm gene inhibit the mycelial growth of phytopathogenic fungi with greater efficiency when compared to the untransformed plants. The transformed and control plants were submitted to salinity resistance test. For this, the plants were maintained in nutrient medium MS with 300 mM NaCl for seven days, after the transformed plants had no difference in growth and number of leaves, however they were able to increase the concentration of free proline when compared to the non-plants transformed. It is possible to measure a probable effect of C. procera latex proteins on the plant defense mechanism against the attack of phytopathogens and to relate the CpOsm protein with mechanisms of action of protection of plants to saline stress and attacks by phytopathogens.
publishDate 2019
dc.date.accessioned.fl_str_mv 2019-09-26T14:17:48Z
dc.date.available.fl_str_mv 2019-09-26T14:17:48Z
dc.date.issued.fl_str_mv 2019
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dc.identifier.citation.fl_str_mv SOUZA, Isabel Cristina da Costa. Análise do envolvimento de proteínas do látex de Calotropis procera na defesa vegetal face a estresses biótico e abiótico. 2019. 119f. Tese (Doutorado em Bioquímica) - Universidade Federal do Ceará, Fortaleza, 2019
dc.identifier.uri.fl_str_mv http://www.repositorio.ufc.br/handle/riufc/46145
identifier_str_mv SOUZA, Isabel Cristina da Costa. Análise do envolvimento de proteínas do látex de Calotropis procera na defesa vegetal face a estresses biótico e abiótico. 2019. 119f. Tese (Doutorado em Bioquímica) - Universidade Federal do Ceará, Fortaleza, 2019
url http://www.repositorio.ufc.br/handle/riufc/46145
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