Mecanismos de defesa da soja [Glycine max (L.) Merril] em diferentes fases de desenvolvimento contra o fungo Cercospora kikuchii e caracterização da toxicidade in vitro da SBTX como fungicida

Detalhes bibliográficos
Ano de defesa: 2020
Autor(a) principal: Arantes, Mariana Reis
Orientador(a): Oliveira, José Tadeu Abreu de
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/59725
Resumo: Soy emerges as one of the main monocultures that drives Brazilian economy. Thus, several strategies are used to reduce the losses that plague this commodity from planting to harvest. Fungi are the main pathogens that cause diseases in plants and several species are routinely studied to create methods and products that will reduce and eradicate the damage caused by these diseases. The necrotrophic fungus Cercospora kikuchii affects aerial parts of soy at the end of the cycle, causing Cercospora leaf blight and purple seed stain and the symptoms usually appear in the later stages of plant growth at the beginning of grain set and can culminate in early defoliation of the upper leaves. The soybean toxin (SBTX) was isolated from soybean seeds and displays antifungal activity against filamentous fungi of agronomic interest and its mechanism of action was elucidated by pathogenic yeasts to man. Although SBTX presents potential to promote or increase the resistance of cultivating soy to fungal diseases, information on its safety of use for man is scarce. Therefore, this work aimed to compare the defense response of soybean in the young and adult phase when inoculated with C. kikuchiii and to investigate the effects of SBTX on non-target cells to ensure its safety of use as an antifungal compound. The presence of the fungus was confirmed in both stages of soybean development, however, only in the adult leaves the symptoms were visualized, the production of cercosporin by the fungus detected and the enhance of CFP expression (Cercosporin Facilitator Protein). Young plants exhibited an increase in the enzymatic activity of superoxide dismutase, as well as more pronounced increases in peroxidase and catalase, resulting in a microenvironment with low production of H2O2. In addition, young plants were still able to enhance PR proteins synthesis earlier when compared to adult plants. Regarding the safety of use of SBTX assessment, it was not able to inhibit the growth of bacteria Salmonella enterica, Bacillus subtilis and Staphylococcus aureus. Furthermore, SBTX did not affect the viability of erythrocytes, neutrophils and Caco-2 cells. However, it altered the gene expression profile of Caco-2 cells after 24 h of exposure, with an upregulation of genes involved in the cell cycle pathway and immune system responses, whereas the genes involved in cholesterol synthesis were downregulated. Thus, despite SBTX did not exhibit negative effects for bacteria or cytotoxicity in mammalian cells, it altered gene expression of some metabolic pathways of Caco-2 cells, indicating that further analysis should be conducted to warrant the safe use of this protein.
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spelling Arantes, Mariana ReisVasconcelos, Ilka MariaOliveira, José Tadeu Abreu de2021-07-26T12:22:21Z2021-07-26T12:22:21Z2020ARANTES, Mariana Reis. Mecanismos de defesa da soja [Glycine max (L.) Merril] em diferentes fases de desenvolvimento contra o fungo Cercospora kikuchii e caracterização da toxicidade in vitro da SBTX como fungicida. 2020. 99 f. Tese (Doutorado em Bioquímica) - Universidade Federal do Ceará, Fortaleza, 2020.http://www.repositorio.ufc.br/handle/riufc/59725Soy emerges as one of the main monocultures that drives Brazilian economy. Thus, several strategies are used to reduce the losses that plague this commodity from planting to harvest. Fungi are the main pathogens that cause diseases in plants and several species are routinely studied to create methods and products that will reduce and eradicate the damage caused by these diseases. The necrotrophic fungus Cercospora kikuchii affects aerial parts of soy at the end of the cycle, causing Cercospora leaf blight and purple seed stain and the symptoms usually appear in the later stages of plant growth at the beginning of grain set and can culminate in early defoliation of the upper leaves. The soybean toxin (SBTX) was isolated from soybean seeds and displays antifungal activity against filamentous fungi of agronomic interest and its mechanism of action was elucidated by pathogenic yeasts to man. Although SBTX presents potential to promote or increase the resistance of cultivating soy to fungal diseases, information on its safety of use for man is scarce. Therefore, this work aimed to compare the defense response of soybean in the young and adult phase when inoculated with C. kikuchiii and to investigate the effects of SBTX on non-target cells to ensure its safety of use as an antifungal compound. The presence of the fungus was confirmed in both stages of soybean development, however, only in the adult leaves the symptoms were visualized, the production of cercosporin by the fungus detected and the enhance of CFP expression (Cercosporin Facilitator Protein). Young plants exhibited an increase in the enzymatic activity of superoxide dismutase, as well as more pronounced increases in peroxidase and catalase, resulting in a microenvironment with low production of H2O2. In addition, young plants were still able to enhance PR proteins synthesis earlier when compared to adult plants. Regarding the safety of use of SBTX assessment, it was not able to inhibit the growth of bacteria Salmonella enterica, Bacillus subtilis and Staphylococcus aureus. Furthermore, SBTX did not affect the viability of erythrocytes, neutrophils and Caco-2 cells. However, it altered the gene expression profile of Caco-2 cells after 24 h of exposure, with an upregulation of genes involved in the cell cycle pathway and immune system responses, whereas the genes involved in cholesterol synthesis were downregulated. Thus, despite SBTX did not exhibit negative effects for bacteria or cytotoxicity in mammalian cells, it altered gene expression of some metabolic pathways of Caco-2 cells, indicating that further analysis should be conducted to warrant the safe use of this protein.A soja desponta como uma das principais monoculturas que movimenta a economia brasileira. Assim, diversas estratégias são utilizadas para reduzir as perdas que assolam essa commodity desde seu plantio à colheita. Os fungos se destacam como principais patógenos causadores de doenças nas plantas e diversas espécies destes microganismos são estudadas rotineiramente com o objetivo de se criar métodos e produtos que venham a minimizar e erradicar os prejuízos causados por essas doenças. O fungo necrotrófico Cercospora kikuchii afeta as partes aéreas da soja no final do seu ciclo de desenvolvimento, causando doenças da queima das folhas e da mancha roxa das sementes. Os sintomas geralmente aparecem em estágios posteriores do crescimento das plantas no início do enchimento dos grãos e podem culminar no desfolhamento precoce das folhas superiores. A toxina da soja (SBTX) foi isolada de sementes de soja apresentando atividade antifúngica contra fungos filamentosos de interesse agronômico e teve seu mecanismo de ação elucidado para leveduras patogênicas ao homem. Apesar da SBTX apresentar características que a elencam como uma ferramenta potencial capaz de promover o aumento da resistência de cultivares de soja a doenças fúngicas, informações acerca da sua segurança de uso para o homem são escassas. Diante disso, os objetivos deste trabalho foram comparar a resposta de defesa da soja desafiadas com C. kikuchii na fase jovem e adulta e investigar a segurança de uso de SBTX como um composto antifúngico. A presença do fungo foi confirmada em ambas as fases de desenvolvimento da soja, contudo, apenas em folhas adultas foram visualizados os sintomas da doença, a produção de cercosporina pelo fungo bem como o aumento da expressão gênica da CFP (Cercosporin Facilitator Protein). A fase jovem apresentou um discreto aumento na atividade enzimática da superoxido dismutase bem como aumentos mais pronunciados da ascorbato peroxidase e catalase resultando em um microambiente com pouca produção de H2O2. Além disso, a fase jovem foi ainda capaz de aumentar a síntese de PR-proteinas de forma mais precoce quando comparada com a fase adulta. Quanto à análise da segurança de uso da SBTX, não foi capaz de inibir o crescimento das bactérias Salmonella enterica, Bacillus subtilis e Staphylococcus aureus. Ainda, SBTX não afetou a viabilidade de eritrócitos, neutrófilos e células de Caco-2. Todavia, SBTX alterou o perfil de expressão gênica de células Caco-2 após 24 h de exposição, havendo a superexpressão de genes envolvidos nas vias do ciclo celular e da resposta do sistema imune, ao passo que genes envolvidos na síntese do colesterol foram subexpressos. Assim, SBTX não apresentou efeitos negativos para bactérias nem induziu citotoxicidade em células de mamíferos, alterou a expressão gênica de algumas vias metabólicas de células de Caco-2, indicando que mais análises devem ser conduzidas para assegurar o uso dessa proteína.Cercospora kikuchiiSBTXsojatoxicogenômicaCaco-2Mecanismos de defesa da soja [Glycine max (L.) Merril] em diferentes fases de desenvolvimento contra o fungo Cercospora kikuchii e caracterização da toxicidade in vitro da SBTX como fungicidaDefense mechanisms of soybean [Glycine max (L.) Merril] in different development phases against Cercospora kikuchii and in vitro toxicity characterization of SBTX as fungicideinfo: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/openAccessLICENSElicense.txtlicense.txttext/plain; charset=utf-81748http://repositorio.ufc.br/bitstream/riufc/59725/6/license.txt8a4605be74aa9ea9d79846c1fba20a33MD56ORIGINAL2020_tese_mrarantes.pdf2020_tese_mrarantes.pdfapplication/pdf2098072http://repositorio.ufc.br/bitstream/riufc/59725/5/2020_tese_mrarantes.pdf424779823334fb17aa10c601ce2f1447MD55riufc/597252021-07-26 09:22:21.353oai:repositorio.ufc.br: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Repositório InstitucionalPUBhttp://www.repositorio.ufc.br/ri-oai/requestbu@ufc.br || repositorio@ufc.bropendoar:2021-07-26T12:22:21Repositório Institucional da Universidade Federal do Ceará (UFC) - Universidade Federal do Ceará (UFC)false
dc.title.pt_BR.fl_str_mv Mecanismos de defesa da soja [Glycine max (L.) Merril] em diferentes fases de desenvolvimento contra o fungo Cercospora kikuchii e caracterização da toxicidade in vitro da SBTX como fungicida
dc.title.en.pt_BR.fl_str_mv Defense mechanisms of soybean [Glycine max (L.) Merril] in different development phases against Cercospora kikuchii and in vitro toxicity characterization of SBTX as fungicide
title Mecanismos de defesa da soja [Glycine max (L.) Merril] em diferentes fases de desenvolvimento contra o fungo Cercospora kikuchii e caracterização da toxicidade in vitro da SBTX como fungicida
spellingShingle Mecanismos de defesa da soja [Glycine max (L.) Merril] em diferentes fases de desenvolvimento contra o fungo Cercospora kikuchii e caracterização da toxicidade in vitro da SBTX como fungicida
Arantes, Mariana Reis
Cercospora kikuchii
SBTX
soja
toxicogenômica
Caco-2
title_short Mecanismos de defesa da soja [Glycine max (L.) Merril] em diferentes fases de desenvolvimento contra o fungo Cercospora kikuchii e caracterização da toxicidade in vitro da SBTX como fungicida
title_full Mecanismos de defesa da soja [Glycine max (L.) Merril] em diferentes fases de desenvolvimento contra o fungo Cercospora kikuchii e caracterização da toxicidade in vitro da SBTX como fungicida
title_fullStr Mecanismos de defesa da soja [Glycine max (L.) Merril] em diferentes fases de desenvolvimento contra o fungo Cercospora kikuchii e caracterização da toxicidade in vitro da SBTX como fungicida
title_full_unstemmed Mecanismos de defesa da soja [Glycine max (L.) Merril] em diferentes fases de desenvolvimento contra o fungo Cercospora kikuchii e caracterização da toxicidade in vitro da SBTX como fungicida
title_sort Mecanismos de defesa da soja [Glycine max (L.) Merril] em diferentes fases de desenvolvimento contra o fungo Cercospora kikuchii e caracterização da toxicidade in vitro da SBTX como fungicida
author Arantes, Mariana Reis
author_facet Arantes, Mariana Reis
author_role author
dc.contributor.co-advisor.none.fl_str_mv Vasconcelos, Ilka Maria
dc.contributor.author.fl_str_mv Arantes, Mariana Reis
dc.contributor.advisor1.fl_str_mv Oliveira, José Tadeu Abreu de
contributor_str_mv Oliveira, José Tadeu Abreu de
dc.subject.por.fl_str_mv Cercospora kikuchii
SBTX
soja
toxicogenômica
Caco-2
topic Cercospora kikuchii
SBTX
soja
toxicogenômica
Caco-2
description Soy emerges as one of the main monocultures that drives Brazilian economy. Thus, several strategies are used to reduce the losses that plague this commodity from planting to harvest. Fungi are the main pathogens that cause diseases in plants and several species are routinely studied to create methods and products that will reduce and eradicate the damage caused by these diseases. The necrotrophic fungus Cercospora kikuchii affects aerial parts of soy at the end of the cycle, causing Cercospora leaf blight and purple seed stain and the symptoms usually appear in the later stages of plant growth at the beginning of grain set and can culminate in early defoliation of the upper leaves. The soybean toxin (SBTX) was isolated from soybean seeds and displays antifungal activity against filamentous fungi of agronomic interest and its mechanism of action was elucidated by pathogenic yeasts to man. Although SBTX presents potential to promote or increase the resistance of cultivating soy to fungal diseases, information on its safety of use for man is scarce. Therefore, this work aimed to compare the defense response of soybean in the young and adult phase when inoculated with C. kikuchiii and to investigate the effects of SBTX on non-target cells to ensure its safety of use as an antifungal compound. The presence of the fungus was confirmed in both stages of soybean development, however, only in the adult leaves the symptoms were visualized, the production of cercosporin by the fungus detected and the enhance of CFP expression (Cercosporin Facilitator Protein). Young plants exhibited an increase in the enzymatic activity of superoxide dismutase, as well as more pronounced increases in peroxidase and catalase, resulting in a microenvironment with low production of H2O2. In addition, young plants were still able to enhance PR proteins synthesis earlier when compared to adult plants. Regarding the safety of use of SBTX assessment, it was not able to inhibit the growth of bacteria Salmonella enterica, Bacillus subtilis and Staphylococcus aureus. Furthermore, SBTX did not affect the viability of erythrocytes, neutrophils and Caco-2 cells. However, it altered the gene expression profile of Caco-2 cells after 24 h of exposure, with an upregulation of genes involved in the cell cycle pathway and immune system responses, whereas the genes involved in cholesterol synthesis were downregulated. Thus, despite SBTX did not exhibit negative effects for bacteria or cytotoxicity in mammalian cells, it altered gene expression of some metabolic pathways of Caco-2 cells, indicating that further analysis should be conducted to warrant the safe use of this protein.
publishDate 2020
dc.date.issued.fl_str_mv 2020
dc.date.accessioned.fl_str_mv 2021-07-26T12:22:21Z
dc.date.available.fl_str_mv 2021-07-26T12:22:21Z
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dc.identifier.citation.fl_str_mv ARANTES, Mariana Reis. Mecanismos de defesa da soja [Glycine max (L.) Merril] em diferentes fases de desenvolvimento contra o fungo Cercospora kikuchii e caracterização da toxicidade in vitro da SBTX como fungicida. 2020. 99 f. Tese (Doutorado em Bioquímica) - Universidade Federal do Ceará, Fortaleza, 2020.
dc.identifier.uri.fl_str_mv http://www.repositorio.ufc.br/handle/riufc/59725
identifier_str_mv ARANTES, Mariana Reis. Mecanismos de defesa da soja [Glycine max (L.) Merril] em diferentes fases de desenvolvimento contra o fungo Cercospora kikuchii e caracterização da toxicidade in vitro da SBTX como fungicida. 2020. 99 f. Tese (Doutorado em Bioquímica) - Universidade Federal do Ceará, Fortaleza, 2020.
url http://www.repositorio.ufc.br/handle/riufc/59725
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