Estudos de expressão em genes potencialmente envolvidos no processo reprodutivo em Anastrepha obliqua (Diptera, Tephritidae)

Detalhes bibliográficos
Ano de defesa: 2014
Autor(a) principal: Nakamura, Aline Minali
Orientador(a): Brito, Reinaldo Alves de lattes
Banca de defesa: Não Informado pela instituição
Tipo de documento: Dissertação
Tipo de acesso: Acesso aberto
Idioma: por
Instituição de defesa: Universidade Federal de São Carlos
Câmpus São Carlos
Programa de Pós-Graduação: Programa de Pós-Graduação em Genética Evolutiva e Biologia Molecular - PPGGEv
Departamento: Não Informado pela instituição
País: Não Informado pela instituição
Palavras-chave em Português:
Palavras-chave em Inglês:
Área do conhecimento CNPq:
Link de acesso: https://repositorio.ufscar.br/handle/20.500.14289/7325
Resumo: The majority of species in the genus Anastrepha is endemic to the Neotropics and many are of great economic importance because they inflict great damage to several different fruit crops. Brazil has low insertion in international markets because of the demand for quality products with no pesticide residues, which force the improvement of control techniques. Thus, an alternative to insect pests control is the Sterile Insect Technique (SIT), which reduces the pest population by mass release of sterile insects. However, sterilization by radiation has many side effects, reducing the competitiveness of released males relative to wild males. For this reason, different strategies have been used, such as the production of transgenic individuals not only in order to sterility but also increasing the vitality presented by these males compared to wild males. To accomplish that, an important approach could be the study of genes involved in the reproductive process. In this work, we selected nine candidate genes for expression studies of our transcriptome data, which have the potential of being involved in the reproductive process in Anastrepha because they belong to gene families that have already been associated to the reproductive process, and showed differential expression between the contrast of virgin and post-mating of transcriptome data in Anastrepha flies. Since no qPCR study has been done to date in Anastrepha, we tested several reference genes for normalization of expression data. The genes Rpl18, Rps17 and Ef1a were deemed suitable and used here to standardize studies of expression between life stages of A. obliqua. The qPCR gene expression analysis revealed interesting gene expression patterns for AttA, Obp56a and Obp99c, which increases the potential of these genes being involved in the reproductive process. Obp56a showed a higher expression in virgin females in contrast to postmating, whereas AttA and Obp99c showed higher expression in male in contrast to females, which may be interesting for genetic control techniques. We applied the RNAi silencing technique with AttA and Obp99c, aiming to generate information about the participation of these genes in reproduction. Thus, our results pointed for three candidate genes that could be interesting for population control techniques, with potential of being involved in the reproductive process, which stimulates further researches in Anastrepha obliqua.
id SCAR_57712e917b2d667204d458f76d4fab4d
oai_identifier_str oai:repositorio.ufscar.br:20.500.14289/7325
network_acronym_str SCAR
network_name_str Repositório Institucional da UFSCAR
repository_id_str
spelling Nakamura, Aline MinaliBrito, Reinaldo Alves dehttp://lattes.cnpq.br/8253066295947754http://lattes.cnpq.br/6716341388414833a03ddecd-67bb-4022-a539-60d78401e45b2016-09-21T13:00:12Z2016-09-21T13:00:12Z2014-10-31NAKAMURA, Aline Minali. Estudos de expressão em genes potencialmente envolvidos no processo reprodutivo em Anastrepha obliqua (Diptera, Tephritidae). 2014. Dissertação (Mestrado em Genética Evolutiva e Biologia Molecular) – Universidade Federal de São Carlos, São Carlos, 2014. Disponível em: https://repositorio.ufscar.br/handle/20.500.14289/7325.https://repositorio.ufscar.br/handle/20.500.14289/7325The majority of species in the genus Anastrepha is endemic to the Neotropics and many are of great economic importance because they inflict great damage to several different fruit crops. Brazil has low insertion in international markets because of the demand for quality products with no pesticide residues, which force the improvement of control techniques. Thus, an alternative to insect pests control is the Sterile Insect Technique (SIT), which reduces the pest population by mass release of sterile insects. However, sterilization by radiation has many side effects, reducing the competitiveness of released males relative to wild males. For this reason, different strategies have been used, such as the production of transgenic individuals not only in order to sterility but also increasing the vitality presented by these males compared to wild males. To accomplish that, an important approach could be the study of genes involved in the reproductive process. In this work, we selected nine candidate genes for expression studies of our transcriptome data, which have the potential of being involved in the reproductive process in Anastrepha because they belong to gene families that have already been associated to the reproductive process, and showed differential expression between the contrast of virgin and post-mating of transcriptome data in Anastrepha flies. Since no qPCR study has been done to date in Anastrepha, we tested several reference genes for normalization of expression data. The genes Rpl18, Rps17 and Ef1a were deemed suitable and used here to standardize studies of expression between life stages of A. obliqua. The qPCR gene expression analysis revealed interesting gene expression patterns for AttA, Obp56a and Obp99c, which increases the potential of these genes being involved in the reproductive process. Obp56a showed a higher expression in virgin females in contrast to postmating, whereas AttA and Obp99c showed higher expression in male in contrast to females, which may be interesting for genetic control techniques. We applied the RNAi silencing technique with AttA and Obp99c, aiming to generate information about the participation of these genes in reproduction. Thus, our results pointed for three candidate genes that could be interesting for population control techniques, with potential of being involved in the reproductive process, which stimulates further researches in Anastrepha obliqua.A maioria das espécies do gênero Anastrepha é endêmica à região neotropical e diversas têm importância econômica por causar grandes prejuízos às culturas de frutos. O Brasil tem baixa inserção no mercado internacional de frutas frescas devido às exigências por produtos de qualidade e sem resíduos de agrotóxicos, o que força o aprimoramento das técnicas de controle de insetospraga. Uma das alternativas é a Técnica do Inseto Estéril (SIT), que visa à redução da população da praga pela liberação em massa de insetos estéreis. Porém, a esterilização por radiação traz muitos efeitos colaterais, diminuindo a competitividade dos machos liberados em relação aos machos selvagens. Por esse motivo, diferentes estratégias têm sido utilizadas como a produção de indivíduos transgênicos visando não só a esterilidade como também o aumento do vigor apresentado por esses machos em relação aos selvagens. Para isso, uma abordagem importante seria o estudo de genes envolvidos no processo reprodutivo. Neste trabalho selecionamos nove genes para estudos de expressão por qPCR candidatos de nossos dados de transcriptomas com potencial de estarem envolvidos no processo reprodutivo em Anastrepha por pertencerem a famílias já relacionadas ao processo reprodutivo, além de apresentarem expressão diferencial entre os transcriptomas de machos virgens e machos pós-cópula. Como nenhum estudo de qPCR havia sido feito ainda em Anastrepha, testamos diversos genes de referência para a normalização dos dados de expressão. Os genes rpl18, rps17 e ef1a foram considerados adequados e padronizados para estudos de expressão entre fases de vida de A. obliqua. As análises de expressão por qPCR revelaram que os genes AttA, Obp56a e Obp99c apresentam padrões de expressão interessantes para investigação e aumentam o potencial desses genes estarem de fato participando do processo reprodutivo. Obp56a apresentou maior expressão em fêmeas virgens em relação às pós-cópula. Já AttA e Obp99c apresentaram maior expressão em machos em relação às fêmeas, fato que pode ser interessante para técnicas de controle genético. Aplicamos a técnica de silenciamento por interferência por RNA nesses genes, com o objetivo de trazer informações sobre a participação desses genes na reprodução. Dessa forma nossos resultados apontam para três genes candidatos que podem ser interessantes para técnicas de controle de populações, com potencial de estar participando do processo reprodutivo o que estimula futuras investigações destes em Anastrepha obliqua.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)porUniversidade Federal de São CarlosCâmpus São CarlosPrograma de Pós-Graduação em Genética Evolutiva e Biologia Molecular - PPGGEvUFSCarAnastrepha obliquaqPCRExpressão diferencialControle genéticoAnastrepha obliquaDifferential expressionGenetic controlCIENCIAS BIOLOGICASEstudos de expressão em genes potencialmente envolvidos no processo reprodutivo em Anastrepha obliqua (Diptera, Tephritidae)info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisOnline600600e4ebf16c-a933-4cae-b428-d05e5cc0dc96info:eu-repo/semantics/openAccessreponame:Repositório Institucional da UFSCARinstname:Universidade Federal de São Carlos (UFSCAR)instacron:UFSCARORIGINALDissAMN.pdfDissAMN.pdfapplication/pdf2525250https://repositorio.ufscar.br/bitstreams/610f7dd7-fc6f-4bcc-bc5b-8609b29803cf/downloadd1ef9cb6cbb03453941fe9349c5da36aMD51trueAnonymousREADLICENSElicense.txtlicense.txttext/plain; charset=utf-81957https://repositorio.ufscar.br/bitstreams/73abb396-7431-48fb-b90a-12ed190252f9/downloadae0398b6f8b235e40ad82cba6c50031dMD52falseAnonymousREADTEXTDissAMN.pdf.txtDissAMN.pdf.txtExtracted texttext/plain247131https://repositorio.ufscar.br/bitstreams/0d647920-5f5d-45c5-a8a8-e89fc7450a72/downloadaa84b46d5c82fd1068bb30243335350dMD55falseAnonymousREADTHUMBNAILDissAMN.pdf.jpgDissAMN.pdf.jpgIM Thumbnailimage/jpeg4342https://repositorio.ufscar.br/bitstreams/dd6d7b05-c512-4262-a6cc-84dd7dd80f16/download7aa4103a8f62882b99005079934ef4b4MD56falseAnonymousREAD20.500.14289/73252025-02-05 17:12:37.559Acesso abertoopen.accessoai:repositorio.ufscar.br:20.500.14289/7325https://repositorio.ufscar.brRepositório InstitucionalPUBhttps://repositorio.ufscar.br/oai/requestrepositorio.sibi@ufscar.bropendoar:43222025-02-05T20:12:37Repositório Institucional da UFSCAR - Universidade Federal de São Carlos (UFSCAR)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
dc.title.por.fl_str_mv Estudos de expressão em genes potencialmente envolvidos no processo reprodutivo em Anastrepha obliqua (Diptera, Tephritidae)
title Estudos de expressão em genes potencialmente envolvidos no processo reprodutivo em Anastrepha obliqua (Diptera, Tephritidae)
spellingShingle Estudos de expressão em genes potencialmente envolvidos no processo reprodutivo em Anastrepha obliqua (Diptera, Tephritidae)
Nakamura, Aline Minali
Anastrepha obliqua
qPCR
Expressão diferencial
Controle genético
Anastrepha obliqua
Differential expression
Genetic control
CIENCIAS BIOLOGICAS
title_short Estudos de expressão em genes potencialmente envolvidos no processo reprodutivo em Anastrepha obliqua (Diptera, Tephritidae)
title_full Estudos de expressão em genes potencialmente envolvidos no processo reprodutivo em Anastrepha obliqua (Diptera, Tephritidae)
title_fullStr Estudos de expressão em genes potencialmente envolvidos no processo reprodutivo em Anastrepha obliqua (Diptera, Tephritidae)
title_full_unstemmed Estudos de expressão em genes potencialmente envolvidos no processo reprodutivo em Anastrepha obliqua (Diptera, Tephritidae)
title_sort Estudos de expressão em genes potencialmente envolvidos no processo reprodutivo em Anastrepha obliqua (Diptera, Tephritidae)
author Nakamura, Aline Minali
author_facet Nakamura, Aline Minali
author_role author
dc.contributor.authorlattes.por.fl_str_mv http://lattes.cnpq.br/6716341388414833
dc.contributor.author.fl_str_mv Nakamura, Aline Minali
dc.contributor.advisor1.fl_str_mv Brito, Reinaldo Alves de
dc.contributor.advisor1Lattes.fl_str_mv http://lattes.cnpq.br/8253066295947754
dc.contributor.authorID.fl_str_mv a03ddecd-67bb-4022-a539-60d78401e45b
contributor_str_mv Brito, Reinaldo Alves de
dc.subject.por.fl_str_mv Anastrepha obliqua
qPCR
Expressão diferencial
Controle genético
topic Anastrepha obliqua
qPCR
Expressão diferencial
Controle genético
Anastrepha obliqua
Differential expression
Genetic control
CIENCIAS BIOLOGICAS
dc.subject.eng.fl_str_mv Anastrepha obliqua
Differential expression
Genetic control
dc.subject.cnpq.fl_str_mv CIENCIAS BIOLOGICAS
description The majority of species in the genus Anastrepha is endemic to the Neotropics and many are of great economic importance because they inflict great damage to several different fruit crops. Brazil has low insertion in international markets because of the demand for quality products with no pesticide residues, which force the improvement of control techniques. Thus, an alternative to insect pests control is the Sterile Insect Technique (SIT), which reduces the pest population by mass release of sterile insects. However, sterilization by radiation has many side effects, reducing the competitiveness of released males relative to wild males. For this reason, different strategies have been used, such as the production of transgenic individuals not only in order to sterility but also increasing the vitality presented by these males compared to wild males. To accomplish that, an important approach could be the study of genes involved in the reproductive process. In this work, we selected nine candidate genes for expression studies of our transcriptome data, which have the potential of being involved in the reproductive process in Anastrepha because they belong to gene families that have already been associated to the reproductive process, and showed differential expression between the contrast of virgin and post-mating of transcriptome data in Anastrepha flies. Since no qPCR study has been done to date in Anastrepha, we tested several reference genes for normalization of expression data. The genes Rpl18, Rps17 and Ef1a were deemed suitable and used here to standardize studies of expression between life stages of A. obliqua. The qPCR gene expression analysis revealed interesting gene expression patterns for AttA, Obp56a and Obp99c, which increases the potential of these genes being involved in the reproductive process. Obp56a showed a higher expression in virgin females in contrast to postmating, whereas AttA and Obp99c showed higher expression in male in contrast to females, which may be interesting for genetic control techniques. We applied the RNAi silencing technique with AttA and Obp99c, aiming to generate information about the participation of these genes in reproduction. Thus, our results pointed for three candidate genes that could be interesting for population control techniques, with potential of being involved in the reproductive process, which stimulates further researches in Anastrepha obliqua.
publishDate 2014
dc.date.issued.fl_str_mv 2014-10-31
dc.date.accessioned.fl_str_mv 2016-09-21T13:00:12Z
dc.date.available.fl_str_mv 2016-09-21T13:00:12Z
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.citation.fl_str_mv NAKAMURA, Aline Minali. Estudos de expressão em genes potencialmente envolvidos no processo reprodutivo em Anastrepha obliqua (Diptera, Tephritidae). 2014. Dissertação (Mestrado em Genética Evolutiva e Biologia Molecular) – Universidade Federal de São Carlos, São Carlos, 2014. Disponível em: https://repositorio.ufscar.br/handle/20.500.14289/7325.
dc.identifier.uri.fl_str_mv https://repositorio.ufscar.br/handle/20.500.14289/7325
identifier_str_mv NAKAMURA, Aline Minali. Estudos de expressão em genes potencialmente envolvidos no processo reprodutivo em Anastrepha obliqua (Diptera, Tephritidae). 2014. Dissertação (Mestrado em Genética Evolutiva e Biologia Molecular) – Universidade Federal de São Carlos, São Carlos, 2014. Disponível em: https://repositorio.ufscar.br/handle/20.500.14289/7325.
url https://repositorio.ufscar.br/handle/20.500.14289/7325
dc.language.iso.fl_str_mv por
language por
dc.relation.confidence.fl_str_mv 600
600
dc.relation.authority.fl_str_mv e4ebf16c-a933-4cae-b428-d05e5cc0dc96
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.publisher.none.fl_str_mv Universidade Federal de São Carlos
Câmpus São Carlos
dc.publisher.program.fl_str_mv Programa de Pós-Graduação em Genética Evolutiva e Biologia Molecular - PPGGEv
dc.publisher.initials.fl_str_mv UFSCar
publisher.none.fl_str_mv Universidade Federal de São Carlos
Câmpus São Carlos
dc.source.none.fl_str_mv reponame:Repositório Institucional da UFSCAR
instname:Universidade Federal de São Carlos (UFSCAR)
instacron:UFSCAR
instname_str Universidade Federal de São Carlos (UFSCAR)
instacron_str UFSCAR
institution UFSCAR
reponame_str Repositório Institucional da UFSCAR
collection Repositório Institucional da UFSCAR
bitstream.url.fl_str_mv https://repositorio.ufscar.br/bitstreams/610f7dd7-fc6f-4bcc-bc5b-8609b29803cf/download
https://repositorio.ufscar.br/bitstreams/73abb396-7431-48fb-b90a-12ed190252f9/download
https://repositorio.ufscar.br/bitstreams/0d647920-5f5d-45c5-a8a8-e89fc7450a72/download
https://repositorio.ufscar.br/bitstreams/dd6d7b05-c512-4262-a6cc-84dd7dd80f16/download
bitstream.checksum.fl_str_mv d1ef9cb6cbb03453941fe9349c5da36a
ae0398b6f8b235e40ad82cba6c50031d
aa84b46d5c82fd1068bb30243335350d
7aa4103a8f62882b99005079934ef4b4
bitstream.checksumAlgorithm.fl_str_mv MD5
MD5
MD5
MD5
repository.name.fl_str_mv Repositório Institucional da UFSCAR - Universidade Federal de São Carlos (UFSCAR)
repository.mail.fl_str_mv repositorio.sibi@ufscar.br
_version_ 1851688827206238208