Isolamento e caracterização de genes da família SnRK e sua expressão em resposta ao déficit hídrico em Coffea canephora

Detalhes bibliográficos
Autor(a) principal: Paiva, Rita Márcia Cardoso de
Data de Publicação: 2009
Tipo de documento: Dissertação
Idioma: por
Título da fonte: LOCUS Repositório Institucional da UFV
Texto Completo: http://locus.ufv.br/handle/123456789/4344
Resumo: Drought tolerance can result from numerous anatomical, morphological and physiological characteristics that can be from constituent or inductive nature. As one of the most important phytohormones known, abscisic acid (ABA) have been studied extensively and several of these studies it has been demonstrating that ABA is responsible for several responses adaptatives in the water deficit. This way, the isolation, caracterization and expression studies of genes that participate in signalling pathway in response to ABA, can supply important data for understanding its role and importance in the mechanism of drought tolerance in coffee. The aims of this work was identifies, in the bank of the coffee, genes belonging to the SnRK family besides verifying the responses in leaves and roots of clones of Coffea canephora, in conditions of severe and moderate water deficit. Plants of the clones 120 (drought tolerant genotype) and 109A (drought sensitive genotype) were cultivated in vases of 12 L and the water deficit was reached by suspending the irrigation until the plants reached -1,5 MPa and -3,0 MPa predawn hydric potential. Homology analyses, allowed the identification of 24 contigs that can represent possible genes of SnRK family in coffee, being 3 contigs of the subfamily SnRK1, 5 contigs in the subfamily SnRK2 and 16 sequences in the subgroup SnRK3. Among the identified sequences, 2 contigs, 15120 and 12770, in this investigation, we found that the ESTs of 15120 and 12770 correspond to different regions of the same gene, homologous to Arabidopsis SnRK2.6 and named cnSnRK2.6. A complete sequence of this gene that was put efficiently in vector of transformation of plants. The promoter sequence of this gene was also identified. Expression analyses, by Real Time quantitative PCR, showed that the expression patterns of some genes of this family are altered in ix conditions of water deficit, in leaves and roots of coffee. In leaves, the genes SnRK2.4, CIPK6 and CIPK9 were induced, while the genes SnRK2.3, CIPK3, CIPK8, CIPK10 and CIPK11 were repressed by the water deficit. However, in roots, the genes SnRK2.3, SnRK2.8, CIPK6, CIPK8 and CIPK9 were induced, what demonstrates that some genes presented a different behavior in those two organs. Furthermore, SnRK genes are also involved in the response to other abiotic stresses. In this study, we also verified the expression of these genes in response to the ABA, osmotic stress and saline stress. For that, a new experimental approach was tested, with use of outstanding leaves of coffee, that a fast and efficient form to studies in coffee. The obtained results indicated a reduction in estomatical aperture (gs) that was accompanied of reductions in liquid carbon assimilation (A) for the treatments with ABA and mannitol, while in the controls treatment any alteration was observed. Gene expression analyses demonstrated that many SnRKs genes are responsive to ABA, osmotic stress and saline stress. SnRK2.6 was induced by ABA and mannitol, but not by salinity. Collectively, these results indicate that the SnRKs characterized genes consist of kinases proteins involved in response to different abiotic stresses.
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spelling Paiva, Rita Márcia Cardoso dehttp://lattes.cnpq.br/3293525698179689Almeida, Andréa Miyasaka dehttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4792501H4Damatta, Fábio Murilohttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4784185Y9Loureiro, Marcelo Ehlershttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4780851Y3Otoni, Wagner Camposhttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4786133Y6Fietto, Luciano Gomeshttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4763824H8Diola, Valdirhttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4133062U6Campos, André Narvaes da Rochahttp://lattes.cnpq.br/47183891618445702015-03-26T13:36:45Z2014-08-142015-03-26T13:36:45Z2009-09-14PAIVA, Rita Márcia Cardoso de. Isolation and characterization of genes SnRK family and his expression in response to the water deficit in Coffea canephora. 2009. 83 f. Dissertação (Mestrado em Controle da maturação e senescência em órgãos perecíveis; Fisiologia molecular de plantas superiores) - Universidade Federal de Viçosa, Viçosa, 2009.http://locus.ufv.br/handle/123456789/4344Drought tolerance can result from numerous anatomical, morphological and physiological characteristics that can be from constituent or inductive nature. As one of the most important phytohormones known, abscisic acid (ABA) have been studied extensively and several of these studies it has been demonstrating that ABA is responsible for several responses adaptatives in the water deficit. This way, the isolation, caracterization and expression studies of genes that participate in signalling pathway in response to ABA, can supply important data for understanding its role and importance in the mechanism of drought tolerance in coffee. The aims of this work was identifies, in the bank of the coffee, genes belonging to the SnRK family besides verifying the responses in leaves and roots of clones of Coffea canephora, in conditions of severe and moderate water deficit. Plants of the clones 120 (drought tolerant genotype) and 109A (drought sensitive genotype) were cultivated in vases of 12 L and the water deficit was reached by suspending the irrigation until the plants reached -1,5 MPa and -3,0 MPa predawn hydric potential. Homology analyses, allowed the identification of 24 contigs that can represent possible genes of SnRK family in coffee, being 3 contigs of the subfamily SnRK1, 5 contigs in the subfamily SnRK2 and 16 sequences in the subgroup SnRK3. Among the identified sequences, 2 contigs, 15120 and 12770, in this investigation, we found that the ESTs of 15120 and 12770 correspond to different regions of the same gene, homologous to Arabidopsis SnRK2.6 and named cnSnRK2.6. A complete sequence of this gene that was put efficiently in vector of transformation of plants. The promoter sequence of this gene was also identified. Expression analyses, by Real Time quantitative PCR, showed that the expression patterns of some genes of this family are altered in ix conditions of water deficit, in leaves and roots of coffee. In leaves, the genes SnRK2.4, CIPK6 and CIPK9 were induced, while the genes SnRK2.3, CIPK3, CIPK8, CIPK10 and CIPK11 were repressed by the water deficit. However, in roots, the genes SnRK2.3, SnRK2.8, CIPK6, CIPK8 and CIPK9 were induced, what demonstrates that some genes presented a different behavior in those two organs. Furthermore, SnRK genes are also involved in the response to other abiotic stresses. In this study, we also verified the expression of these genes in response to the ABA, osmotic stress and saline stress. For that, a new experimental approach was tested, with use of outstanding leaves of coffee, that a fast and efficient form to studies in coffee. The obtained results indicated a reduction in estomatical aperture (gs) that was accompanied of reductions in liquid carbon assimilation (A) for the treatments with ABA and mannitol, while in the controls treatment any alteration was observed. Gene expression analyses demonstrated that many SnRKs genes are responsive to ABA, osmotic stress and saline stress. SnRK2.6 was induced by ABA and mannitol, but not by salinity. Collectively, these results indicate that the SnRKs characterized genes consist of kinases proteins involved in response to different abiotic stresses.A tolerância à seca é o resultado de numerosas características anatômicas, morfológicas e fisiológicas, de natureza constitutiva ou indutiva. Como um dos principais hormônios de plantas, o ácido abscísico (ABA) tem sido extensivamente estudado e vários destes estudos têm demonstrado que o ABA é responsável por diversas respostas adaptativas ao déficit hídrico. Desta forma, o isolamento e estudo da expressão de genes que participam de vias de sinalização em resposta a ABA podem fornecer dados importantes para a compreensão de sua função e de sua importância no mecanismo de tolerância à seca em café. Este trabalho teve como objetivos identificar, a partir do banco do café, genes pertencentes à família SnRK além de verificar a resposta dos mesmos em folhas e raízes de clones de Coffea canephora, em condições de déficit hídrico severo e moderado. Mudas dos clones 120 (genótipo tolerante ao déficit hídrico) e 109A (genótipo sensível) foram cultivadas em vasos de 12 L e o déficit hídrico foi alcançado suspendendo a irrigação até que as plantas atingissem um potencial hídrico de antemanhã de -1,5 MPa e -3,0 MPa. Análises de homologia utilizando o programa Blast, permitiram a identificação de 24 contigs que podem representar possíveis genes da família SnRK em café, sendo 3 contigs da subfamília SnRK1, 5 contigs pertencentes a subfamília SnRK2 e 16 sequências da subfamília SnRK3. Dentre as sequências identificadas, destacam-se 2 contigs, 15120 e 12770, que nesta investigação, demonstrou-se que correspondem a regiões distintas de um mesmo gene, homólogo a SnRK2.6 de Arabidopsis e denominado cnSnRK2.6. Desta forma, foi obtida a sequência completa deste gene que foi eficientemente colocado em vetor de transformação de plantas. Neste estudo, identificou-se também uma parte da região promotora deste gene. Análises de vii expressão, feitas por RT-PCR em Tempo Real, permitiu verificar que a expressão de alguns genes desta família são alteradas em condições de déficit hídrico, tanto em folhas como em raízes. Em folhas, os genes SnRK2.4, CIPK6 e CIPK9 são induzidos, enquanto os genes SnRK2.3, CIPK3, CIPK8, CIPK10 e CIPK11 são reprimidos pela condição de seca. Já em raízes, observou-se que os genes SnRK2.3, SnRK2.8, CIPK6, CIPK8 e CIPK9 são induzidos, o que demonstra que alguns genes apresentaram comportamento diferenciado nesses dois órgãos. Em adição, foi também verificado a expressão destes genes em resposta à ABA, estresse osmótico e estresse salino. Para isso, foi testado um novo sistema experimental, com uso de folhas destacadas de cafeeiro, que se mostrou uma forma rápida e eficiente para estudos em plantas de café. Os resultados obtidos mostraram uma redução da condutância estomática (gs) que foi acompanhada de reduções nas taxas de assimilação líquida do carbono (A) para os tratamentos com ABA e manitol, enquanto no tratamento controle nenhuma alteração foi observada. A análise de expressão gênica, demonstrou que muitos desses genes respondem a ABA, estresse osmótico e estresse salino. SnRK2.6 é induzido por ABA e manitol, mas não por salinidade. Coletivamente, estes resultados sugerem que os genes SnRKs caracterizados constituem proteínas quinases envolvidas na resposta a estresses abióticos.Conselho Nacional de Desenvolvimento Científico e Tecnológicoapplication/pdfporUniversidade Federal de ViçosaMestrado em Fisiologia VegetalUFVBRControle da maturação e senescência em órgãos perecíveis; Fisiologia molecular de plantas superioresEstresse hídricoCoffea canephoraSnRKWater stressCoffea canephoraSnRKCNPQ::CIENCIAS AGRARIAS::AGRONOMIA::FITOTECNIA::FISIOLOGIA DE PLANTAS CULTIVADASIsolamento e caracterização de genes da família SnRK e sua expressão em resposta ao déficit hídrico em Coffea canephoraIsolation and characterization of genes SnRK family and his expression in response to the water deficit in Coffea canephorainfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisinfo:eu-repo/semantics/openAccessreponame:LOCUS Repositório Institucional da UFVinstname:Universidade Federal de Viçosa (UFV)instacron:UFVORIGINALtexto completo.pdfapplication/pdf794761https://locus.ufv.br//bitstream/123456789/4344/1/texto%20completo.pdf2c6f4b2d5ee311e882270c54be62a38cMD51TEXTtexto completo.pdf.txttexto completo.pdf.txtExtracted texttext/plain141082https://locus.ufv.br//bitstream/123456789/4344/2/texto%20completo.pdf.txt13acafdb38029b01b204f2547ab6ddfaMD52THUMBNAILtexto completo.pdf.jpgtexto completo.pdf.jpgIM Thumbnailimage/jpeg3702https://locus.ufv.br//bitstream/123456789/4344/3/texto%20completo.pdf.jpg0c4157d4f605ce40b4af71b84a1e9930MD53123456789/43442016-04-10 23:08:03.598oai:locus.ufv.br:123456789/4344Repositório InstitucionalPUBhttps://www.locus.ufv.br/oai/requestfabiojreis@ufv.bropendoar:21452016-04-11T02:08:03LOCUS Repositório Institucional da UFV - Universidade Federal de Viçosa (UFV)false
dc.title.por.fl_str_mv Isolamento e caracterização de genes da família SnRK e sua expressão em resposta ao déficit hídrico em Coffea canephora
dc.title.alternative.eng.fl_str_mv Isolation and characterization of genes SnRK family and his expression in response to the water deficit in Coffea canephora
title Isolamento e caracterização de genes da família SnRK e sua expressão em resposta ao déficit hídrico em Coffea canephora
spellingShingle Isolamento e caracterização de genes da família SnRK e sua expressão em resposta ao déficit hídrico em Coffea canephora
Paiva, Rita Márcia Cardoso de
Estresse hídrico
Coffea canephora
SnRK
Water stress
Coffea canephora
SnRK
CNPQ::CIENCIAS AGRARIAS::AGRONOMIA::FITOTECNIA::FISIOLOGIA DE PLANTAS CULTIVADAS
title_short Isolamento e caracterização de genes da família SnRK e sua expressão em resposta ao déficit hídrico em Coffea canephora
title_full Isolamento e caracterização de genes da família SnRK e sua expressão em resposta ao déficit hídrico em Coffea canephora
title_fullStr Isolamento e caracterização de genes da família SnRK e sua expressão em resposta ao déficit hídrico em Coffea canephora
title_full_unstemmed Isolamento e caracterização de genes da família SnRK e sua expressão em resposta ao déficit hídrico em Coffea canephora
title_sort Isolamento e caracterização de genes da família SnRK e sua expressão em resposta ao déficit hídrico em Coffea canephora
author Paiva, Rita Márcia Cardoso de
author_facet Paiva, Rita Márcia Cardoso de
author_role author
dc.contributor.authorLattes.por.fl_str_mv http://lattes.cnpq.br/3293525698179689
dc.contributor.author.fl_str_mv Paiva, Rita Márcia Cardoso de
dc.contributor.advisor-co1.fl_str_mv Almeida, Andréa Miyasaka de
dc.contributor.advisor-co1Lattes.fl_str_mv http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4792501H4
dc.contributor.advisor-co2.fl_str_mv Damatta, Fábio Murilo
dc.contributor.advisor-co2Lattes.fl_str_mv http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4784185Y9
dc.contributor.advisor1.fl_str_mv Loureiro, Marcelo Ehlers
dc.contributor.advisor1Lattes.fl_str_mv http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4780851Y3
dc.contributor.referee1.fl_str_mv Otoni, Wagner Campos
dc.contributor.referee1Lattes.fl_str_mv http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4786133Y6
dc.contributor.referee2.fl_str_mv Fietto, Luciano Gomes
dc.contributor.referee2Lattes.fl_str_mv http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4763824H8
dc.contributor.referee3.fl_str_mv Diola, Valdir
dc.contributor.referee3Lattes.fl_str_mv http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4133062U6
dc.contributor.referee4.fl_str_mv Campos, André Narvaes da Rocha
dc.contributor.referee4Lattes.fl_str_mv http://lattes.cnpq.br/4718389161844570
contributor_str_mv Almeida, Andréa Miyasaka de
Damatta, Fábio Murilo
Loureiro, Marcelo Ehlers
Otoni, Wagner Campos
Fietto, Luciano Gomes
Diola, Valdir
Campos, André Narvaes da Rocha
dc.subject.por.fl_str_mv Estresse hídrico
Coffea canephora
SnRK
topic Estresse hídrico
Coffea canephora
SnRK
Water stress
Coffea canephora
SnRK
CNPQ::CIENCIAS AGRARIAS::AGRONOMIA::FITOTECNIA::FISIOLOGIA DE PLANTAS CULTIVADAS
dc.subject.eng.fl_str_mv Water stress
Coffea canephora
SnRK
dc.subject.cnpq.fl_str_mv CNPQ::CIENCIAS AGRARIAS::AGRONOMIA::FITOTECNIA::FISIOLOGIA DE PLANTAS CULTIVADAS
description Drought tolerance can result from numerous anatomical, morphological and physiological characteristics that can be from constituent or inductive nature. As one of the most important phytohormones known, abscisic acid (ABA) have been studied extensively and several of these studies it has been demonstrating that ABA is responsible for several responses adaptatives in the water deficit. This way, the isolation, caracterization and expression studies of genes that participate in signalling pathway in response to ABA, can supply important data for understanding its role and importance in the mechanism of drought tolerance in coffee. The aims of this work was identifies, in the bank of the coffee, genes belonging to the SnRK family besides verifying the responses in leaves and roots of clones of Coffea canephora, in conditions of severe and moderate water deficit. Plants of the clones 120 (drought tolerant genotype) and 109A (drought sensitive genotype) were cultivated in vases of 12 L and the water deficit was reached by suspending the irrigation until the plants reached -1,5 MPa and -3,0 MPa predawn hydric potential. Homology analyses, allowed the identification of 24 contigs that can represent possible genes of SnRK family in coffee, being 3 contigs of the subfamily SnRK1, 5 contigs in the subfamily SnRK2 and 16 sequences in the subgroup SnRK3. Among the identified sequences, 2 contigs, 15120 and 12770, in this investigation, we found that the ESTs of 15120 and 12770 correspond to different regions of the same gene, homologous to Arabidopsis SnRK2.6 and named cnSnRK2.6. A complete sequence of this gene that was put efficiently in vector of transformation of plants. The promoter sequence of this gene was also identified. Expression analyses, by Real Time quantitative PCR, showed that the expression patterns of some genes of this family are altered in ix conditions of water deficit, in leaves and roots of coffee. In leaves, the genes SnRK2.4, CIPK6 and CIPK9 were induced, while the genes SnRK2.3, CIPK3, CIPK8, CIPK10 and CIPK11 were repressed by the water deficit. However, in roots, the genes SnRK2.3, SnRK2.8, CIPK6, CIPK8 and CIPK9 were induced, what demonstrates that some genes presented a different behavior in those two organs. Furthermore, SnRK genes are also involved in the response to other abiotic stresses. In this study, we also verified the expression of these genes in response to the ABA, osmotic stress and saline stress. For that, a new experimental approach was tested, with use of outstanding leaves of coffee, that a fast and efficient form to studies in coffee. The obtained results indicated a reduction in estomatical aperture (gs) that was accompanied of reductions in liquid carbon assimilation (A) for the treatments with ABA and mannitol, while in the controls treatment any alteration was observed. Gene expression analyses demonstrated that many SnRKs genes are responsive to ABA, osmotic stress and saline stress. SnRK2.6 was induced by ABA and mannitol, but not by salinity. Collectively, these results indicate that the SnRKs characterized genes consist of kinases proteins involved in response to different abiotic stresses.
publishDate 2009
dc.date.issued.fl_str_mv 2009-09-14
dc.date.available.fl_str_mv 2014-08-14
2015-03-26T13:36:45Z
dc.date.accessioned.fl_str_mv 2015-03-26T13:36:45Z
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dc.identifier.citation.fl_str_mv PAIVA, Rita Márcia Cardoso de. Isolation and characterization of genes SnRK family and his expression in response to the water deficit in Coffea canephora. 2009. 83 f. Dissertação (Mestrado em Controle da maturação e senescência em órgãos perecíveis; Fisiologia molecular de plantas superiores) - Universidade Federal de Viçosa, Viçosa, 2009.
dc.identifier.uri.fl_str_mv http://locus.ufv.br/handle/123456789/4344
identifier_str_mv PAIVA, Rita Márcia Cardoso de. Isolation and characterization of genes SnRK family and his expression in response to the water deficit in Coffea canephora. 2009. 83 f. Dissertação (Mestrado em Controle da maturação e senescência em órgãos perecíveis; Fisiologia molecular de plantas superiores) - Universidade Federal de Viçosa, Viçosa, 2009.
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