Caracterização do exsudato de sementes de Moringa oleífera Lamarck e investigação de seu papel na defesa do vegetal.
Autor(a) principal: | |
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Data de Publicação: | 2013 |
Tipo de documento: | Dissertação |
Idioma: | por |
Título da fonte: | Repositório Institucional da Universidade Federal do Ceará (UFC) |
Texto Completo: | http://www.repositorio.ufc.br/handle/riufc/9655 |
Resumo: | Moringa oleifera (moringa) is a species belonging to the family Moringaceae which is characterized as having high resistance to insects and fungi. Previous work carried out by our research group revealed the presence of chitin-binding proteins in moringa seeds, among them Mo-CBP3, suggesting a positive correlation between this protein and the plant resistance. At the onset of the new plant development, for germination to occur, seed imbibition is required, a process followed by exudation. In exudation process, primary and secondary metabolites are released in the medium outside the seeds, some of them protecting the new plant against herbivores and/or pathogens. This study aimed to characterize the chemical composition and biological activities of moringa seed exudate and to investigate the presence of Mo-CBP3 in the exudate, in order to contribute to the establishment of its physiological role. Initially, the best conditions for exudation were established, emphasizing the time and solvent. A higher exudation of seed proteins was observed in distilled water after 24 hours. This exudate showed the presence of activities related to primary (protease, β-1,3- glucanase, chitinase, trypsin inhibitor and papain inhibitor) and secondary (steroid and saponins) metabolites. Mo-CBP3 was also detected in the exudate, using polyclonal antibodies anti-Mo-CBP3. The presence of Mo-CBP3 in the moringa seed exudate was confirmed after chromatography on chitin matrix and analyses by dot blotting and polyacrylamide gel electrophoresis. The data obtained showed that the retained material on the chitin matrix corresponds to 0.26% of the total protein, it is recognized by anti-Mo-CBP3 and has electrophoretic profile similar to that of Mo- CBP3 which was purified from moringa seeds. In the activity tests to pathogens, the seed exudate showed no antifungal activity, under the conditions used, except for Candida parapsilosis which had a slight reduction in its growth rate. In contrast, a potent activity against nematode was found as the seed exudate was able to cause a mortality rate up to 100% of Meloidogyne incognita in J2 stage. When investigated the presence of Mo-CBP3 in moringa hoots, a plant organ that shows the exudation phenomen and is also able to interact directly with the nematode, positive results were found. In fact, Mo-CBP3 is present in moringa hoots, in the initial stages of the plant development, according to the results shown by ELISA and RT-PCR. The data, taken together, suggest that in the exudation phenomen, proteins must play essential roles. In the case of moringa, Mo-CBP3 already participates in the initial stages of development of this tree species, playing a role that must be related to protection against pathogens. |
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Caracterização do exsudato de sementes de Moringa oleífera Lamarck e investigação de seu papel na defesa do vegetal.Characterization of seed exudate Moringa oleifera Lamarck and investigation of their role in plant defense.BioquímicaMoringa oleiferaExsudação (botânica)Moringa oleifera (moringa) is a species belonging to the family Moringaceae which is characterized as having high resistance to insects and fungi. Previous work carried out by our research group revealed the presence of chitin-binding proteins in moringa seeds, among them Mo-CBP3, suggesting a positive correlation between this protein and the plant resistance. At the onset of the new plant development, for germination to occur, seed imbibition is required, a process followed by exudation. In exudation process, primary and secondary metabolites are released in the medium outside the seeds, some of them protecting the new plant against herbivores and/or pathogens. This study aimed to characterize the chemical composition and biological activities of moringa seed exudate and to investigate the presence of Mo-CBP3 in the exudate, in order to contribute to the establishment of its physiological role. Initially, the best conditions for exudation were established, emphasizing the time and solvent. A higher exudation of seed proteins was observed in distilled water after 24 hours. This exudate showed the presence of activities related to primary (protease, β-1,3- glucanase, chitinase, trypsin inhibitor and papain inhibitor) and secondary (steroid and saponins) metabolites. Mo-CBP3 was also detected in the exudate, using polyclonal antibodies anti-Mo-CBP3. The presence of Mo-CBP3 in the moringa seed exudate was confirmed after chromatography on chitin matrix and analyses by dot blotting and polyacrylamide gel electrophoresis. The data obtained showed that the retained material on the chitin matrix corresponds to 0.26% of the total protein, it is recognized by anti-Mo-CBP3 and has electrophoretic profile similar to that of Mo- CBP3 which was purified from moringa seeds. In the activity tests to pathogens, the seed exudate showed no antifungal activity, under the conditions used, except for Candida parapsilosis which had a slight reduction in its growth rate. In contrast, a potent activity against nematode was found as the seed exudate was able to cause a mortality rate up to 100% of Meloidogyne incognita in J2 stage. When investigated the presence of Mo-CBP3 in moringa hoots, a plant organ that shows the exudation phenomen and is also able to interact directly with the nematode, positive results were found. In fact, Mo-CBP3 is present in moringa hoots, in the initial stages of the plant development, according to the results shown by ELISA and RT-PCR. The data, taken together, suggest that in the exudation phenomen, proteins must play essential roles. In the case of moringa, Mo-CBP3 already participates in the initial stages of development of this tree species, playing a role that must be related to protection against pathogens.Moringa oleifera (moringa) é uma espécie pertencente à família Moringaceae que se caracteriza por ser muito resistente a insetos e fungos. Trabalhos prévios realizados por nosso grupo de pesquisa revelaram a presença de proteínas ligantes à quitina em sementes de moringa, dentre elas a Mo-CBP3, sugerindo uma correlação positiva entre essa proteína e a resistência da planta. No início do desenvolvimento da planta, para que ocorra a germinação, deve haver a embebição da semente, um processo seguido pela exsudação. Na exsudação, compostos do metabolismo primário e secundário são externalizados da semente, alguns deles exercendo ação de defesa da nova planta ao impedirem o ataque de herbívoros e/ou patógenos. O presente trabalho teve como objetivos caracterizar o exsudato de sementes de moringa quanto à composição bioquímica e atividade biológica e investigar a presença de Mo-CBP3 no exsudato, visando contribuir para o estabelecimento de seu papel fisiológico. Inicialmente, foram estabelecidas as condições de exsudação, dando ênfase ao tempo e solvente. Uma maior exsudação de proteínas foi observada em sementes embebidas com água destilada por 24 horas. Esse exsudato mostrou a presença de atividades inerentes a metabólitos primários (protease, β-1,3-glucanase, quitinase, inibidor de tripsina e inibidor de papaína) e secundários (saponinas e esteroides). Mo-CBP3 foi também detectada no exsudato, usando anticorpos policlonais anti-Mo-CBP3. A presença de Mo-CBP3 no exsudato de sementes de moringa foi confirmada após este ter sido submetido à cromatografia em matriz de quitina, procedida pela análise através de dot bloting e eletroforese em gel de poliacrilamida. Os dados obtidos mostraram que o material retido na matriz de quitina corresponde a 0,26% do total de proteínas exsudadas, é reconhecido pelo anticorpo anti-Mo-CBP3 e apresenta perfil eletroforético similar ao da Mo-CBP3 purificada de sementes de moringa. Na avaliação da atividade deste exsudato frente à fitopatógenos, ação contra fungos não foi detectada, nas condições de ensaio empregadas, exceto para Candida parapsilosis que mostrou uma discreta redução na taxa de crescimento. Contrariamente, uma potente atividade contra nematoide foi verificada, tendo sido o exsudato de sementes capaz de causar até 100% de mortalidade para indivíduos de Meloidogyne incognita em estágio de J2. Quando investigada a presença de Mo-CBP3 em raiz, um órgão vegetal que apresenta o fenômeno de exsudação e, também, é capaz de interagir diretamente com o nematoide, resultados positivos foram encontrados. De fato, Mo- CBP3 está presente em raízes de moringa, já nos estágios iniciais do desenvolvimento, conforme resultados mostrados por ELISA e através da técnica de RT-PCR. Os dados, em conjunto, sugerem que no fenômeno da exsudação, proteínas devem desempenhar funções essenciais e que, no caso da moringa, Mo- CBP3 já participa nos estágios iniciais do desenvolvimento dessa planta arbórea, papel este que pode estar relacionado com a proteção contra patógenos.Vasconcelos, Ilka MariaSousa, Antônio Juscelino Sudário2014-11-03T20:09:22Z2014-11-03T20:09:22Z2013info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisapplication/pdfSOUSA, A. J. S. Caracterização do exsudato de sementes de Moringa oleífera Lamarck e investigação de seu papel na defesa do vegetal. 2013. 103 f. Dissertação (Mestrado em Bioquímica) - Centro de Ciências, Universidade Federal do Ceará, Fortaleza, 2013.http://www.repositorio.ufc.br/handle/riufc/9655porreponame:Repositório Institucional da Universidade Federal do Ceará (UFC)instname:Universidade Federal do Ceará (UFC)instacron:UFCinfo:eu-repo/semantics/openAccess2020-05-22T16:52:05Zoai:repositorio.ufc.br:riufc/9655Repositório InstitucionalPUBhttp://www.repositorio.ufc.br/ri-oai/requestbu@ufc.br || repositorio@ufc.bropendoar:2024-09-11T18:24:29.359501Repositório Institucional da Universidade Federal do Ceará (UFC) - Universidade Federal do Ceará (UFC)false |
dc.title.none.fl_str_mv |
Caracterização do exsudato de sementes de Moringa oleífera Lamarck e investigação de seu papel na defesa do vegetal. Characterization of seed exudate Moringa oleifera Lamarck and investigation of their role in plant defense. |
title |
Caracterização do exsudato de sementes de Moringa oleífera Lamarck e investigação de seu papel na defesa do vegetal. |
spellingShingle |
Caracterização do exsudato de sementes de Moringa oleífera Lamarck e investigação de seu papel na defesa do vegetal. Sousa, Antônio Juscelino Sudário Bioquímica Moringa oleifera Exsudação (botânica) |
title_short |
Caracterização do exsudato de sementes de Moringa oleífera Lamarck e investigação de seu papel na defesa do vegetal. |
title_full |
Caracterização do exsudato de sementes de Moringa oleífera Lamarck e investigação de seu papel na defesa do vegetal. |
title_fullStr |
Caracterização do exsudato de sementes de Moringa oleífera Lamarck e investigação de seu papel na defesa do vegetal. |
title_full_unstemmed |
Caracterização do exsudato de sementes de Moringa oleífera Lamarck e investigação de seu papel na defesa do vegetal. |
title_sort |
Caracterização do exsudato de sementes de Moringa oleífera Lamarck e investigação de seu papel na defesa do vegetal. |
author |
Sousa, Antônio Juscelino Sudário |
author_facet |
Sousa, Antônio Juscelino Sudário |
author_role |
author |
dc.contributor.none.fl_str_mv |
Vasconcelos, Ilka Maria |
dc.contributor.author.fl_str_mv |
Sousa, Antônio Juscelino Sudário |
dc.subject.por.fl_str_mv |
Bioquímica Moringa oleifera Exsudação (botânica) |
topic |
Bioquímica Moringa oleifera Exsudação (botânica) |
description |
Moringa oleifera (moringa) is a species belonging to the family Moringaceae which is characterized as having high resistance to insects and fungi. Previous work carried out by our research group revealed the presence of chitin-binding proteins in moringa seeds, among them Mo-CBP3, suggesting a positive correlation between this protein and the plant resistance. At the onset of the new plant development, for germination to occur, seed imbibition is required, a process followed by exudation. In exudation process, primary and secondary metabolites are released in the medium outside the seeds, some of them protecting the new plant against herbivores and/or pathogens. This study aimed to characterize the chemical composition and biological activities of moringa seed exudate and to investigate the presence of Mo-CBP3 in the exudate, in order to contribute to the establishment of its physiological role. Initially, the best conditions for exudation were established, emphasizing the time and solvent. A higher exudation of seed proteins was observed in distilled water after 24 hours. This exudate showed the presence of activities related to primary (protease, β-1,3- glucanase, chitinase, trypsin inhibitor and papain inhibitor) and secondary (steroid and saponins) metabolites. Mo-CBP3 was also detected in the exudate, using polyclonal antibodies anti-Mo-CBP3. The presence of Mo-CBP3 in the moringa seed exudate was confirmed after chromatography on chitin matrix and analyses by dot blotting and polyacrylamide gel electrophoresis. The data obtained showed that the retained material on the chitin matrix corresponds to 0.26% of the total protein, it is recognized by anti-Mo-CBP3 and has electrophoretic profile similar to that of Mo- CBP3 which was purified from moringa seeds. In the activity tests to pathogens, the seed exudate showed no antifungal activity, under the conditions used, except for Candida parapsilosis which had a slight reduction in its growth rate. In contrast, a potent activity against nematode was found as the seed exudate was able to cause a mortality rate up to 100% of Meloidogyne incognita in J2 stage. When investigated the presence of Mo-CBP3 in moringa hoots, a plant organ that shows the exudation phenomen and is also able to interact directly with the nematode, positive results were found. In fact, Mo-CBP3 is present in moringa hoots, in the initial stages of the plant development, according to the results shown by ELISA and RT-PCR. The data, taken together, suggest that in the exudation phenomen, proteins must play essential roles. In the case of moringa, Mo-CBP3 already participates in the initial stages of development of this tree species, playing a role that must be related to protection against pathogens. |
publishDate |
2013 |
dc.date.none.fl_str_mv |
2013 2014-11-03T20:09:22Z 2014-11-03T20:09:22Z |
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 |
SOUSA, A. J. S. Caracterização do exsudato de sementes de Moringa oleífera Lamarck e investigação de seu papel na defesa do vegetal. 2013. 103 f. Dissertação (Mestrado em Bioquímica) - Centro de Ciências, Universidade Federal do Ceará, Fortaleza, 2013. http://www.repositorio.ufc.br/handle/riufc/9655 |
identifier_str_mv |
SOUSA, A. J. S. Caracterização do exsudato de sementes de Moringa oleífera Lamarck e investigação de seu papel na defesa do vegetal. 2013. 103 f. Dissertação (Mestrado em Bioquímica) - Centro de Ciências, Universidade Federal do Ceará, Fortaleza, 2013. |
url |
http://www.repositorio.ufc.br/handle/riufc/9655 |
dc.language.iso.fl_str_mv |
por |
language |
por |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
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openAccess |
dc.format.none.fl_str_mv |
application/pdf |
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reponame:Repositório Institucional da Universidade Federal do Ceará (UFC) instname:Universidade Federal do Ceará (UFC) instacron:UFC |
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Universidade Federal do Ceará (UFC) |
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UFC |
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UFC |
reponame_str |
Repositório Institucional da Universidade Federal do Ceará (UFC) |
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Repositório Institucional da Universidade Federal do Ceará (UFC) |
repository.name.fl_str_mv |
Repositório Institucional da Universidade Federal do Ceará (UFC) - Universidade Federal do Ceará (UFC) |
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bu@ufc.br || repositorio@ufc.br |
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