Plantas transgênicas de tabaco expressando RNA hairpin e jaburetox como estratégias para controle de Helicoverpa armigera

Detalhes bibliográficos
Autor(a) principal: Ceccon, Cássia Canzi
Data de Publicação: 2019
Tipo de documento: Tese
Idioma: por
Título da fonte: Biblioteca de teses e dissertações da Universidade de Passo Fundo (BDTD UPF)
Texto Completo: http://tede.upf.br:8080/jspui/handle/tede/1765
Resumo: Helicoverpa armigera is a serious insect-pest in several regions of the world and there are reports of populations resistant to various groups of currently used insecticides and Bt toxins. Considering these factors, the need to search for new H. armigera control strategies is clear. Technologies such as oral delivery of RNA interference (RNAi) inducing molecules that induce mortality by silencing vital target pest genes and the use of new insecticidal biomolecules, such as the Jaburetox (JBTX) recombinant insecticidal peptide, derived from one of the Canavalia ensiformes urease isoforms, are strategies that can be used to control pest insect, including H. armigera. The objective of this work was to produce genetically modified tobacco plants containing genes expressing: 1) hairpin RNA molecules (hpRNA) that form doble- strand RNA (dsRNA) complementary to the H. armigera rieske gene (hpRNA-rieske); and/or, 2) the entomotoxic Jaburetox peptide (JBTX), and evaluate their efficacy in inducing mortality in H. armigera fed on these plants. High mortality and reduced consumption were verified in larvae fed on the R12/2 event containing the gene encoding hpRNA-rieske and on J22/4 event containing the gene encoding the JBTX peptide. Feeding of the larvae with the hpRNA-rieske event resulted in a reduction in the level of the rieske gene transcripts in the digestive system and in the remaining tissues of the larvae body by 85%. Using the best events (R12/2 and J22/4) as parents, progenies of containing both genes of interest were obtained through the crosses J22/4xR12/2 and R12/2xJ22/4. These stacked plants, hemizygous for both genes, induced similar mortality in H. armigera as the R12/2 parent. This parent displays segregation of hpRNA-rieske in offspring suggesting it is homozygous; however, it could also carry more than one copy of hpRNA-rieske as a result of multiple insertions in the genome. It is hypothesized that the evaluation of J22/4 and R12/2 hybrids developed in the future to be homozygous for both genes may provide even higher control of H. armigera. In addition, further studies may indicate the occurrence of interaction between insecticidal substances and the feasibility of using both technologies together in a stacked approach.
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spelling Salvadori, José Roberto18921981004http://lattes.cnpq.br/477215306099062502339906075http://lattes.cnpq.br/3095134137550295Ceccon, Cássia Canzi2019-08-23T19:21:09Z2019-09-09CECCON, Cássia Canzi. Plantas transgênicas de tabaco expressando RNA hairpin e jaburetox como estratégias para controle de Helicoverpa armigera. 2019. 124 f.Dissertação (Mestrado em Agronomia) - Universidade de Passo Fundo, Passo Fundo, RS, 2019.http://tede.upf.br:8080/jspui/handle/tede/1765Helicoverpa armigera is a serious insect-pest in several regions of the world and there are reports of populations resistant to various groups of currently used insecticides and Bt toxins. Considering these factors, the need to search for new H. armigera control strategies is clear. Technologies such as oral delivery of RNA interference (RNAi) inducing molecules that induce mortality by silencing vital target pest genes and the use of new insecticidal biomolecules, such as the Jaburetox (JBTX) recombinant insecticidal peptide, derived from one of the Canavalia ensiformes urease isoforms, are strategies that can be used to control pest insect, including H. armigera. The objective of this work was to produce genetically modified tobacco plants containing genes expressing: 1) hairpin RNA molecules (hpRNA) that form doble- strand RNA (dsRNA) complementary to the H. armigera rieske gene (hpRNA-rieske); and/or, 2) the entomotoxic Jaburetox peptide (JBTX), and evaluate their efficacy in inducing mortality in H. armigera fed on these plants. High mortality and reduced consumption were verified in larvae fed on the R12/2 event containing the gene encoding hpRNA-rieske and on J22/4 event containing the gene encoding the JBTX peptide. Feeding of the larvae with the hpRNA-rieske event resulted in a reduction in the level of the rieske gene transcripts in the digestive system and in the remaining tissues of the larvae body by 85%. Using the best events (R12/2 and J22/4) as parents, progenies of containing both genes of interest were obtained through the crosses J22/4xR12/2 and R12/2xJ22/4. These stacked plants, hemizygous for both genes, induced similar mortality in H. armigera as the R12/2 parent. This parent displays segregation of hpRNA-rieske in offspring suggesting it is homozygous; however, it could also carry more than one copy of hpRNA-rieske as a result of multiple insertions in the genome. It is hypothesized that the evaluation of J22/4 and R12/2 hybrids developed in the future to be homozygous for both genes may provide even higher control of H. armigera. In addition, further studies may indicate the occurrence of interaction between insecticidal substances and the feasibility of using both technologies together in a stacked approach.Helicoverpa armigera se destaca como inseto-praga em diversas regiões do mundo e apresenta populações resistentes a diversos grupos de inseticidas e toxinas Bt utilizados atualmente. Considerando estes fatores, é visível a necessidade da busca por novas estratégias com potencial de controle desta praga. Tecnologias como a disponibilização oral de moléculas de RNA de interferência (RNAi) que induzem a mortalidade e o silenciamento de genes vitais aos insetos-praga alvo e a utilização de novas biomoléculas inseticidas, como o peptídeo recombinante Jaburetox (JBTX), derivado de uma das isoformas de urease de Canavalia ensiformes, são estratégias que podem ser utilizadas para o controle de insetos-praga, incluindo H. armigera. O objetivo deste trabalho foi produzir plantas de tabaco geneticamente modificadas contendo os genes que expressam: 1) moléculas de RNA em forma de grampo (hairpin RNA-hpRNA) que forma a molécula de RNA dupla-fita (RNAdf) complementar ao gene rieske de H. armigera (hpRNA-rieske); e/ou, 2) o peptídeo entomotóxico Jaburetox (JBTX), e avaliar a eficácia na indução de mortalidade de larvas de H. armigera alimentadas com estas plantas. Foi verificado alta mortalidade e redução do consumo nas larvas alimentadas com os eventos R12/2 contendo o gene para a expressão do hpRNA-rieske e J22/4, contendo o gene para expressão do JBTX. A alimentação das larvas com os eventos hpRNA-rieske resultou na redução do nível de transcritos do gene rieske no sistema digestório e nos tecidos do restante do corpo das larvas, chegando a 85% de silenciamento. Utilizando os melhores eventos R12/2 e J22/4 foi obtido progênies de plantas empilhadas contendo ambos os genes de interesse pelos cruzamentos J22/4xR12/2 e R12/2xJ22/4. As plantas empilhadas, heterozigotas para ambos os genes, mostraram resposta semelhante ao parental R12/2. Esse progenitor apresenta segregação do transgenes que sugere que está em homozigose, porém, pode haver mais de uma cópia do gene hpRNA-rieske como resultado de múltiplas inserções no genoma. Hipotetiza-se que a avaliação dos híbridos J22/4 e R12/2 desenvolvidos no futuro para que apresentem homozigose para ambos os genes pode fornecer melhores resultados quanto ao potencial de controle de H. armigera. Além disso, mais estudos poderão indicar a ocorrência de interação entre as substâncias inseticidas e a viabilidade de utilização de ambas tecnologias em conjunto.Submitted by Aline Rezende (alinerezende@upf.br) on 2019-08-23T19:21:09Z No. of bitstreams: 1 2019CassiaCeccon.pdf: 2809779 bytes, checksum: 686beb1d40a175453cc205b35f2e891b (MD5)Made available in DSpace on 2019-08-23T19:21:09Z (GMT). No. of bitstreams: 1 2019CassiaCeccon.pdf: 2809779 bytes, checksum: 686beb1d40a175453cc205b35f2e891b (MD5) Previous issue date: 2019-09-09application/pdfporUniversidade de Passo FundoPrograma de Pós-Graduação em AgronomiaUPFBrasilFaculdade de Agronomia e Medicina Veterinária – FAMVInseto nocivo - ControlePlantas transgênicasFumo - Melhoramento genéticoCIENCIAS AGRARIAS::AGRONOMIAPlantas transgênicas de tabaco expressando RNA hairpin e jaburetox como estratégias para controle de Helicoverpa armigeraTransgenic tobacco plants expressing hairpin and jaburetox RNA as strategies for Helicoverpa armigera controlinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesis562406611703505429050050060053202200503672799-3091138714907603907info:eu-repo/semantics/openAccessreponame:Biblioteca de teses e dissertações da Universidade de Passo Fundo (BDTD UPF)instname:Universidade de Passo Fundo (UPF)instacron:UPFORIGINAL2019CassiaCeccon.pdf2019CassiaCeccon.pdfapplication/pdf2809779http://tede.upf.br:8080/jspui/bitstream/tede/1765/2/2019CassiaCeccon.pdf686beb1d40a175453cc205b35f2e891bMD52LICENSElicense.txtlicense.txttext/plain; charset=utf-82165http://tede.upf.br:8080/jspui/bitstream/tede/1765/1/license.txtbd3efa91386c1718a7f26a329fdcb468MD51tede/17652019-08-23 16:21:09.466oai:tede.upf.br: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Biblioteca Digital de Teses e DissertaçõesPUBhttp://tede.upf.br/oai/requestbiblio@upf.br || bio@upf.br || cas@upf.br || car@upf.br || lve@upf.br || sar@upf.br || sol@upf.br || upfmundi@upf.br || jucelei@upf.bropendoar:2019-08-23T19:21:09Biblioteca de teses e dissertações da Universidade de Passo Fundo (BDTD UPF) - Universidade de Passo Fundo (UPF)false
dc.title.por.fl_str_mv Plantas transgênicas de tabaco expressando RNA hairpin e jaburetox como estratégias para controle de Helicoverpa armigera
dc.title.alternative.eng.fl_str_mv Transgenic tobacco plants expressing hairpin and jaburetox RNA as strategies for Helicoverpa armigera control
title Plantas transgênicas de tabaco expressando RNA hairpin e jaburetox como estratégias para controle de Helicoverpa armigera
spellingShingle Plantas transgênicas de tabaco expressando RNA hairpin e jaburetox como estratégias para controle de Helicoverpa armigera
Ceccon, Cássia Canzi
Inseto nocivo - Controle
Plantas transgênicas
Fumo - Melhoramento genético
CIENCIAS AGRARIAS::AGRONOMIA
title_short Plantas transgênicas de tabaco expressando RNA hairpin e jaburetox como estratégias para controle de Helicoverpa armigera
title_full Plantas transgênicas de tabaco expressando RNA hairpin e jaburetox como estratégias para controle de Helicoverpa armigera
title_fullStr Plantas transgênicas de tabaco expressando RNA hairpin e jaburetox como estratégias para controle de Helicoverpa armigera
title_full_unstemmed Plantas transgênicas de tabaco expressando RNA hairpin e jaburetox como estratégias para controle de Helicoverpa armigera
title_sort Plantas transgênicas de tabaco expressando RNA hairpin e jaburetox como estratégias para controle de Helicoverpa armigera
author Ceccon, Cássia Canzi
author_facet Ceccon, Cássia Canzi
author_role author
dc.contributor.advisor1.fl_str_mv Salvadori, José Roberto
dc.contributor.advisor1ID.fl_str_mv 18921981004
dc.contributor.advisor1Lattes.fl_str_mv http://lattes.cnpq.br/4772153060990625
dc.contributor.authorID.fl_str_mv 02339906075
dc.contributor.authorLattes.fl_str_mv http://lattes.cnpq.br/3095134137550295
dc.contributor.author.fl_str_mv Ceccon, Cássia Canzi
contributor_str_mv Salvadori, José Roberto
dc.subject.por.fl_str_mv Inseto nocivo - Controle
Plantas transgênicas
Fumo - Melhoramento genético
topic Inseto nocivo - Controle
Plantas transgênicas
Fumo - Melhoramento genético
CIENCIAS AGRARIAS::AGRONOMIA
dc.subject.cnpq.fl_str_mv CIENCIAS AGRARIAS::AGRONOMIA
description Helicoverpa armigera is a serious insect-pest in several regions of the world and there are reports of populations resistant to various groups of currently used insecticides and Bt toxins. Considering these factors, the need to search for new H. armigera control strategies is clear. Technologies such as oral delivery of RNA interference (RNAi) inducing molecules that induce mortality by silencing vital target pest genes and the use of new insecticidal biomolecules, such as the Jaburetox (JBTX) recombinant insecticidal peptide, derived from one of the Canavalia ensiformes urease isoforms, are strategies that can be used to control pest insect, including H. armigera. The objective of this work was to produce genetically modified tobacco plants containing genes expressing: 1) hairpin RNA molecules (hpRNA) that form doble- strand RNA (dsRNA) complementary to the H. armigera rieske gene (hpRNA-rieske); and/or, 2) the entomotoxic Jaburetox peptide (JBTX), and evaluate their efficacy in inducing mortality in H. armigera fed on these plants. High mortality and reduced consumption were verified in larvae fed on the R12/2 event containing the gene encoding hpRNA-rieske and on J22/4 event containing the gene encoding the JBTX peptide. Feeding of the larvae with the hpRNA-rieske event resulted in a reduction in the level of the rieske gene transcripts in the digestive system and in the remaining tissues of the larvae body by 85%. Using the best events (R12/2 and J22/4) as parents, progenies of containing both genes of interest were obtained through the crosses J22/4xR12/2 and R12/2xJ22/4. These stacked plants, hemizygous for both genes, induced similar mortality in H. armigera as the R12/2 parent. This parent displays segregation of hpRNA-rieske in offspring suggesting it is homozygous; however, it could also carry more than one copy of hpRNA-rieske as a result of multiple insertions in the genome. It is hypothesized that the evaluation of J22/4 and R12/2 hybrids developed in the future to be homozygous for both genes may provide even higher control of H. armigera. In addition, further studies may indicate the occurrence of interaction between insecticidal substances and the feasibility of using both technologies together in a stacked approach.
publishDate 2019
dc.date.accessioned.fl_str_mv 2019-08-23T19:21:09Z
dc.date.issued.fl_str_mv 2019-09-09
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/doctoralThesis
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dc.identifier.citation.fl_str_mv CECCON, Cássia Canzi. Plantas transgênicas de tabaco expressando RNA hairpin e jaburetox como estratégias para controle de Helicoverpa armigera. 2019. 124 f.Dissertação (Mestrado em Agronomia) - Universidade de Passo Fundo, Passo Fundo, RS, 2019.
dc.identifier.uri.fl_str_mv http://tede.upf.br:8080/jspui/handle/tede/1765
identifier_str_mv CECCON, Cássia Canzi. Plantas transgênicas de tabaco expressando RNA hairpin e jaburetox como estratégias para controle de Helicoverpa armigera. 2019. 124 f.Dissertação (Mestrado em Agronomia) - Universidade de Passo Fundo, Passo Fundo, RS, 2019.
url http://tede.upf.br:8080/jspui/handle/tede/1765
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dc.publisher.department.fl_str_mv Faculdade de Agronomia e Medicina Veterinária – FAMV
publisher.none.fl_str_mv Universidade de Passo Fundo
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