Identificação molecular de helicoverpa armigera (Lepidoptera: Noctuidae)
Autor(a) principal: | |
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Data de Publicação: | 2021 |
Tipo de documento: | Tese |
Idioma: | por |
Título da fonte: | Repositório Institucional da UFG |
Texto Completo: | http://repositorio.bc.ufg.br/tede/handle/tede/12678 |
Resumo: | Previously found only on Old World continents, the caterpillar Helicoverpa armigera Hübner (Lepidoptera: Noctuidae) is capable of great economic damage, coming to be considered one of the most destructive agricultural pests. Its identification through morphological analysis is only possible with complex dissection of the adult male genitalia. The morphology of all phases of the life cycle, egg, larva, pupa and adult, is very similar with the morphology of other species of the genus, making the identification difficult. As it is a highly invasive pest and correct identification is the first step to be taken to decide the best management strategy, we realize the need for precise, easy and fast tools for the identification of this species. Thus, the objective of the work was to develop specific primers and to optimize different protocols for use in real-time PCR and digital PCR methodologies in order to identify H. armigera, even if this sample is of poor quality, only a fragment and/or is mixed with other species. For that, design, selection and optimization of primer and probe concentrations, primer annealing temperature gradients, tests and adaptations with different DNA extraction methods were carried out, with or without post-DNA extraction purification, tests with different ratios of H. zea and H. armigera mixed in the same reaction, determination of the maximum sensitivity in both methodologies and interpretation of the obtained data. We present the protocols developed to successfully identify the specie H. armigera, both with real-time PCR and digital PCR methodologies. |
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Cunha, Marcos Gomes dahttp://lattes.cnpq.br/2006008625763742Tembrock, Luke RaymondGilligan, Todd MichaelCunha, Marcos Gomes daTimm, Alicia EvaDianese, Érico de CamposCoelho, Regina Melo SartoriGodinho, Karina Cordeiro Albernazhttp://lattes.cnpq.br/2051386045777953Oliveira, Thayssa Monize Rosa de2023-03-20T15:48:58Z2023-03-20T15:48:58Z2021-05-21OLIVEIRA, T. M. R. Identificação molecular de helicoverpa armigera (Lepidoptera: Noctuidae). 2021. 75 f. Tese (Doutorado em Agronomia) - Universidade Federal de Goiás, Goiânia, 2021.http://repositorio.bc.ufg.br/tede/handle/tede/12678Previously found only on Old World continents, the caterpillar Helicoverpa armigera Hübner (Lepidoptera: Noctuidae) is capable of great economic damage, coming to be considered one of the most destructive agricultural pests. Its identification through morphological analysis is only possible with complex dissection of the adult male genitalia. The morphology of all phases of the life cycle, egg, larva, pupa and adult, is very similar with the morphology of other species of the genus, making the identification difficult. As it is a highly invasive pest and correct identification is the first step to be taken to decide the best management strategy, we realize the need for precise, easy and fast tools for the identification of this species. Thus, the objective of the work was to develop specific primers and to optimize different protocols for use in real-time PCR and digital PCR methodologies in order to identify H. armigera, even if this sample is of poor quality, only a fragment and/or is mixed with other species. For that, design, selection and optimization of primer and probe concentrations, primer annealing temperature gradients, tests and adaptations with different DNA extraction methods were carried out, with or without post-DNA extraction purification, tests with different ratios of H. zea and H. armigera mixed in the same reaction, determination of the maximum sensitivity in both methodologies and interpretation of the obtained data. We present the protocols developed to successfully identify the specie H. armigera, both with real-time PCR and digital PCR methodologies.Encontrada anteriormente apenas em continentes do Velho Mundo, a lagarta Helicoverpa armigera Hübner (Lepidoptera: Noctuidae) é capaz de grandes danos econômicos, chegando a ser considerada uma das mais destrutivas pragas agrícolas. Sua identificação por meio da análise morfológica só é possível com dissecção complexa da genitália masculina adulta. A morfologia de todas as fases do ciclo de vida, ovo, larva, pupa e adulto, é muito semelhante com a morfologia de outras espécies do gênero, dificultando a identificação. Por ser praga altamente invasiva e observando-se que a correta identificação é o primeiro passo a ser tomado para decisão da melhor estratégia de manejo, percebemos a necessidade de ferramentas precisas, fáceis e rápidas para identificação dessa espécie. Assim, o objetivo do trabalho foi desenvolver primers específicos e otimizar diferentes protocolos para utilização nas metodologias de PCR em tempo real e PCR digital de forma a identificar H. armigera, mesmo que essa amostra seja de má qualidade, apenas um fragmento e/ou esteja misturada com outras espécies. Para tanto, foram realizados desenho, seleção e otimização das concentrações dos primers e sonda, gradientes de temperatura de anelamento dos primers, testes e adaptações com diferentes métodos de extração de DNA, com ou sem purificação pós-extração, testes com diferentes taxas de DNA de H. zea e H. armigera misturados em uma mesma reação, determinação da sensibilidade máxima em ambas as metodologias e interpretação dos dados obtidos. Dessa forma, apresentamos os protocolos desenvolvidos para identificar com sucesso a espécie H. armigera, tanto na metodologia de PCR em tempo real quanto na metodologia de PCR digital.Submitted by Marlene Santos (marlene.bc.ufg@gmail.com) on 2023-03-16T17:51:58Z No. of bitstreams: 2 Tese - Thayssa Monize Rosa de Oliveira - 2021.pdf: 3251620 bytes, checksum: 065fdd1eb7dcea341756b5b8b82ce44c (MD5) license_rdf: 805 bytes, checksum: 4460e5956bc1d1639be9ae6146a50347 (MD5)Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2023-03-20T15:48:57Z (GMT) No. of bitstreams: 2 Tese - Thayssa Monize Rosa de Oliveira - 2021.pdf: 3251620 bytes, checksum: 065fdd1eb7dcea341756b5b8b82ce44c (MD5) license_rdf: 805 bytes, checksum: 4460e5956bc1d1639be9ae6146a50347 (MD5)Made available in DSpace on 2023-03-20T15:48:58Z (GMT). No. of bitstreams: 2 Tese - Thayssa Monize Rosa de Oliveira - 2021.pdf: 3251620 bytes, checksum: 065fdd1eb7dcea341756b5b8b82ce44c (MD5) license_rdf: 805 bytes, checksum: 4460e5956bc1d1639be9ae6146a50347 (MD5) Previous issue date: 2021-05-21Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPESFundação de Amparo à Pesquisa do Estado de GoiásporUniversidade Federal de GoiásPrograma de Pós-graduação em Agronomia (EA)UFGBrasilEscola de Agronomia - EA (RMG)Attribution-NonCommercial-NoDerivatives 4.0 Internationalhttp://creativecommons.org/licenses/by-nc-nd/4.0/info:eu-repo/semantics/openAccessLagarta do velho mundoPCRPrimers específicosOld world bollwormSpecie-specific primersCIENCIAS AGRARIAS::AGRONOMIA::FITOSSANIDADEIdentificação molecular de helicoverpa armigera (Lepidoptera: Noctuidae)Molecular identification of helicoverpa armigera (Lepidoptera: Noctuidae)info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesis550050050050050022213reponame:Repositório Institucional da UFGinstname:Universidade Federal de Goiás (UFG)instacron:UFGLICENSElicense.txtlicense.txttext/plain; charset=utf-81748http://repositorio.bc.ufg.br/tede/bitstreams/be6e9ae0-e212-4ee1-af61-95aaaa121c14/download8a4605be74aa9ea9d79846c1fba20a33MD51CC-LICENSElicense_rdflicense_rdfapplication/rdf+xml; charset=utf-8805http://repositorio.bc.ufg.br/tede/bitstreams/9176f75f-48cc-48f2-b5e2-6ccc7b62bac6/download4460e5956bc1d1639be9ae6146a50347MD52ORIGINALTese - Thayssa Monize Rosa de Oliveira - 2021.pdfTese - Thayssa Monize Rosa de Oliveira - 2021.pdfapplication/pdf3251620http://repositorio.bc.ufg.br/tede/bitstreams/54170bcf-848c-4052-92db-e845fb6fe31d/download065fdd1eb7dcea341756b5b8b82ce44cMD53tede/126782023-03-20 12:48:58.224http://creativecommons.org/licenses/by-nc-nd/4.0/Attribution-NonCommercial-NoDerivatives 4.0 Internationalopen.accessoai:repositorio.bc.ufg.br:tede/12678http://repositorio.bc.ufg.br/tedeRepositório InstitucionalPUBhttp://repositorio.bc.ufg.br/oai/requesttasesdissertacoes.bc@ufg.bropendoar:2023-03-20T15:48:58Repositório Institucional da UFG - Universidade Federal de Goiás (UFG)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 |
dc.title.pt_BR.fl_str_mv |
Identificação molecular de helicoverpa armigera (Lepidoptera: Noctuidae) |
dc.title.alternative.eng.fl_str_mv |
Molecular identification of helicoverpa armigera (Lepidoptera: Noctuidae) |
title |
Identificação molecular de helicoverpa armigera (Lepidoptera: Noctuidae) |
spellingShingle |
Identificação molecular de helicoverpa armigera (Lepidoptera: Noctuidae) Oliveira, Thayssa Monize Rosa de Lagarta do velho mundo PCR Primers específicos Old world bollworm Specie-specific primers CIENCIAS AGRARIAS::AGRONOMIA::FITOSSANIDADE |
title_short |
Identificação molecular de helicoverpa armigera (Lepidoptera: Noctuidae) |
title_full |
Identificação molecular de helicoverpa armigera (Lepidoptera: Noctuidae) |
title_fullStr |
Identificação molecular de helicoverpa armigera (Lepidoptera: Noctuidae) |
title_full_unstemmed |
Identificação molecular de helicoverpa armigera (Lepidoptera: Noctuidae) |
title_sort |
Identificação molecular de helicoverpa armigera (Lepidoptera: Noctuidae) |
author |
Oliveira, Thayssa Monize Rosa de |
author_facet |
Oliveira, Thayssa Monize Rosa de |
author_role |
author |
dc.contributor.advisor1.fl_str_mv |
Cunha, Marcos Gomes da |
dc.contributor.advisor1Lattes.fl_str_mv |
http://lattes.cnpq.br/2006008625763742 |
dc.contributor.advisor-co1.fl_str_mv |
Tembrock, Luke Raymond |
dc.contributor.advisor-co2.fl_str_mv |
Gilligan, Todd Michael |
dc.contributor.referee1.fl_str_mv |
Cunha, Marcos Gomes da |
dc.contributor.referee2.fl_str_mv |
Timm, Alicia Eva |
dc.contributor.referee3.fl_str_mv |
Dianese, Érico de Campos |
dc.contributor.referee4.fl_str_mv |
Coelho, Regina Melo Sartori |
dc.contributor.referee5.fl_str_mv |
Godinho, Karina Cordeiro Albernaz |
dc.contributor.authorLattes.fl_str_mv |
http://lattes.cnpq.br/2051386045777953 |
dc.contributor.author.fl_str_mv |
Oliveira, Thayssa Monize Rosa de |
contributor_str_mv |
Cunha, Marcos Gomes da Tembrock, Luke Raymond Gilligan, Todd Michael Cunha, Marcos Gomes da Timm, Alicia Eva Dianese, Érico de Campos Coelho, Regina Melo Sartori Godinho, Karina Cordeiro Albernaz |
dc.subject.por.fl_str_mv |
Lagarta do velho mundo PCR Primers específicos |
topic |
Lagarta do velho mundo PCR Primers específicos Old world bollworm Specie-specific primers CIENCIAS AGRARIAS::AGRONOMIA::FITOSSANIDADE |
dc.subject.eng.fl_str_mv |
Old world bollworm Specie-specific primers |
dc.subject.cnpq.fl_str_mv |
CIENCIAS AGRARIAS::AGRONOMIA::FITOSSANIDADE |
description |
Previously found only on Old World continents, the caterpillar Helicoverpa armigera Hübner (Lepidoptera: Noctuidae) is capable of great economic damage, coming to be considered one of the most destructive agricultural pests. Its identification through morphological analysis is only possible with complex dissection of the adult male genitalia. The morphology of all phases of the life cycle, egg, larva, pupa and adult, is very similar with the morphology of other species of the genus, making the identification difficult. As it is a highly invasive pest and correct identification is the first step to be taken to decide the best management strategy, we realize the need for precise, easy and fast tools for the identification of this species. Thus, the objective of the work was to develop specific primers and to optimize different protocols for use in real-time PCR and digital PCR methodologies in order to identify H. armigera, even if this sample is of poor quality, only a fragment and/or is mixed with other species. For that, design, selection and optimization of primer and probe concentrations, primer annealing temperature gradients, tests and adaptations with different DNA extraction methods were carried out, with or without post-DNA extraction purification, tests with different ratios of H. zea and H. armigera mixed in the same reaction, determination of the maximum sensitivity in both methodologies and interpretation of the obtained data. We present the protocols developed to successfully identify the specie H. armigera, both with real-time PCR and digital PCR methodologies. |
publishDate |
2021 |
dc.date.issued.fl_str_mv |
2021-05-21 |
dc.date.accessioned.fl_str_mv |
2023-03-20T15:48:58Z |
dc.date.available.fl_str_mv |
2023-03-20T15:48:58Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/doctoralThesis |
format |
doctoralThesis |
status_str |
publishedVersion |
dc.identifier.citation.fl_str_mv |
OLIVEIRA, T. M. R. Identificação molecular de helicoverpa armigera (Lepidoptera: Noctuidae). 2021. 75 f. Tese (Doutorado em Agronomia) - Universidade Federal de Goiás, Goiânia, 2021. |
dc.identifier.uri.fl_str_mv |
http://repositorio.bc.ufg.br/tede/handle/tede/12678 |
identifier_str_mv |
OLIVEIRA, T. M. R. Identificação molecular de helicoverpa armigera (Lepidoptera: Noctuidae). 2021. 75 f. Tese (Doutorado em Agronomia) - Universidade Federal de Goiás, Goiânia, 2021. |
url |
http://repositorio.bc.ufg.br/tede/handle/tede/12678 |
dc.language.iso.fl_str_mv |
por |
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por |
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500 500 500 500 500 |
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2 |
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22 |
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1 3 |
dc.rights.driver.fl_str_mv |
Attribution-NonCommercial-NoDerivatives 4.0 International http://creativecommons.org/licenses/by-nc-nd/4.0/ info:eu-repo/semantics/openAccess |
rights_invalid_str_mv |
Attribution-NonCommercial-NoDerivatives 4.0 International http://creativecommons.org/licenses/by-nc-nd/4.0/ |
eu_rights_str_mv |
openAccess |
dc.publisher.none.fl_str_mv |
Universidade Federal de Goiás |
dc.publisher.program.fl_str_mv |
Programa de Pós-graduação em Agronomia (EA) |
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UFG |
dc.publisher.country.fl_str_mv |
Brasil |
dc.publisher.department.fl_str_mv |
Escola de Agronomia - EA (RMG) |
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Universidade Federal de Goiás |
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