Efeito da superexpress?o do fator de transcri??o OsDof25 sobre a efici?ncia de absor??o de nitrog?nio em Orysa sativa L.

Detalhes bibliográficos
Autor(a) principal: Silva, Renata Aparecida Costa
Data de Publicação: 2012
Tipo de documento: Dissertação
Idioma: por
Título da fonte: Biblioteca Digital de Teses e Dissertações da UFRRJ
Texto Completo: https://tede.ufrrj.br/jspui/handle/jspui/1692
Resumo: Nitrogen is one of the nutrient elements most limiting for plant growth. Thus, increasing plant nitrogen usage efficiency (NUE) is an essential factor for sustainable agriculture, leading to an increased food production with less fertilizer input and less environment impact. The aim of this study was to evaluate the effect of OsDof25 overexpression on N-NO3- and N-NH4+ uptake. In transgenic rice plants, OsDof25 was expressed under control of maize ubiquitin promoter (UBIL:OsDof25:3xHA). Two experiments were conducted: one to evaluate the kinetic parameters Vm?x and KM, and another to analyze the expression level of nitrate (NRT2.1~2.2 and NAR) and ammonium transporters (AMT1.1~1.3), both under high and low NO3- and NH4+ supply. The untransformed plants showed higher growth that transformed lineages. The L1 and L2 showed a lower value of the KM in the resupply treatment of 0.2 mM N-NO3-. In the resupply with 0.2 mM N-NH4 + the L4 showed higher Vmax and L1 lower KM. There were no large variations in uptake kinetics between the transformed and untransformed plants. At the root the NRT2 showed low expression in lineages L1 and L4, when under constant supply of N-NO3-, in contrast, in the treatment under resupply with 0.2 mM N-NO3-was increased expression of OsNTR2.1 ~ 2.2, and NAR in both transformed lineages, but in the root the concentration of NO3- was opposed to the expression of NRT2 and NAR, in both conditions. In the leaves, the line L4 showed high expression of the transporter OsNRT2.1 with the resupply of 0.2 and 2.0 mM N-NO3-. In plants grown under constant supply of N-NH4+, L1 showed lower expression of AMT1 in the root compared to L4 and untransformed plants. When subjected to nitrogen deficiency, there was an increased expression of the OsAMT1.2. However, there was no correlation between N transporter expression levels and NO3- and NH4+ content in the transformed plants, indicating a possible change in enzyme activity and reduction or assimilation of N in these plants. The transformed plants when subjected to resupply with low levels of nitrate and ammonium showed better response parameters Vmax and KM compared to the untransformed. In the plants transformed the resupply with nitrate at low concentration resulted in increasing the gene expression of the transporters (OsNTR2.1 ~ 2.2 and protein OsNAR2.1), and the treatment with constant supply provided greatest nitrate accumulation in these plants. The results of both kinetic parameters and accumulation of fresh matter suggest that plants transformed for the expression of the OsDof25 presented highest tolerance to nutritional stress.
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spelling Souza, Sonia Regina de003.927-487-03Santos, Leandro Azevedo983.907.835-68Souza., Sonia Regina deFerraz J?nior, Altamiro Souza de Lima |Cabral, Luiz Mors077.966.316-00http://lattes.cnpq.br/6396561004465081Silva, Renata Aparecida Costa2017-05-26T20:35:35Z2012-02-15SILVA, Renata Aparecida Costa. Efeito da superexpress?o do fator de transcri??o OsDof25 sobre a efici?ncia de absor??o de nitrog?nio em Orysa sativa L. 2012. 52 f. Disserta??o (Mestrado em Agronomia - Ci?ncia do Solo). Instituto de Agronomia, Departamento de Solos, Universidade Federal Rural do Rio de Janeiro, Serop?dica, 2012.https://tede.ufrrj.br/jspui/handle/jspui/1692Nitrogen is one of the nutrient elements most limiting for plant growth. Thus, increasing plant nitrogen usage efficiency (NUE) is an essential factor for sustainable agriculture, leading to an increased food production with less fertilizer input and less environment impact. The aim of this study was to evaluate the effect of OsDof25 overexpression on N-NO3- and N-NH4+ uptake. In transgenic rice plants, OsDof25 was expressed under control of maize ubiquitin promoter (UBIL:OsDof25:3xHA). Two experiments were conducted: one to evaluate the kinetic parameters Vm?x and KM, and another to analyze the expression level of nitrate (NRT2.1~2.2 and NAR) and ammonium transporters (AMT1.1~1.3), both under high and low NO3- and NH4+ supply. The untransformed plants showed higher growth that transformed lineages. The L1 and L2 showed a lower value of the KM in the resupply treatment of 0.2 mM N-NO3-. In the resupply with 0.2 mM N-NH4 + the L4 showed higher Vmax and L1 lower KM. There were no large variations in uptake kinetics between the transformed and untransformed plants. At the root the NRT2 showed low expression in lineages L1 and L4, when under constant supply of N-NO3-, in contrast, in the treatment under resupply with 0.2 mM N-NO3-was increased expression of OsNTR2.1 ~ 2.2, and NAR in both transformed lineages, but in the root the concentration of NO3- was opposed to the expression of NRT2 and NAR, in both conditions. In the leaves, the line L4 showed high expression of the transporter OsNRT2.1 with the resupply of 0.2 and 2.0 mM N-NO3-. In plants grown under constant supply of N-NH4+, L1 showed lower expression of AMT1 in the root compared to L4 and untransformed plants. When subjected to nitrogen deficiency, there was an increased expression of the OsAMT1.2. However, there was no correlation between N transporter expression levels and NO3- and NH4+ content in the transformed plants, indicating a possible change in enzyme activity and reduction or assimilation of N in these plants. The transformed plants when subjected to resupply with low levels of nitrate and ammonium showed better response parameters Vmax and KM compared to the untransformed. In the plants transformed the resupply with nitrate at low concentration resulted in increasing the gene expression of the transporters (OsNTR2.1 ~ 2.2 and protein OsNAR2.1), and the treatment with constant supply provided greatest nitrate accumulation in these plants. The results of both kinetic parameters and accumulation of fresh matter suggest that plants transformed for the expression of the OsDof25 presented highest tolerance to nutritional stress.O nitrog?nio ? um dos elementos minerais que mais limita o desenvolvimento das plantas. Assim, aumentar a efici?ncia de uso de nitrog?nio (EUN) ? um fator ? essencial para uma agricultura sustent?vel, levando a um aumento da produ??o de alimentos com menor uso de insumos e menos impactos ao ambiente. Este trabalho teve por objetivo avaliar o efeito da superexpress?o do fator de transcri??o OsDof25 sobre a absor??o de nitrog?nio em duas linhagens transformadas de arroz (L1 e L4) da variedade Nipponbare comparando-as com plantas n?o transformadas (WT). Nas plantas transformadas, o OsDof25 foi expresso sob o controle do promotor da ubiquitina 1 de milho (UBIL:OsDof25:3xHA). For feitos dois experimentos: um para avaliar os par?metros cin?ticos Vm?x e KM, sob condi??es de alto e baixo suprimento de N-NO3- e N-NH4+, e outro para analisar a express?o dos transportadores de NO3- (NRT2.1~2.2 e NAR) e NH4+ (AMT1.1~1.3) tamb?m sob alto e baixo suprimento desses ?ons. As plantas n?o transformadas apresentaram maior crescimento do que as linhagens transformadas. As L1 e a L2 mostraram menor valor de KM no tratamento com ressuprimento de 0,2 mM de N-NO3-. No ressuprimento com 0,2 mM de N-NH4+ a L4 apresentou maior Vm?x e L1 menor KM, mas, n?o houve grandes varia??es nos par?metros cin?ticos de absor??o entre as plantas transformadas e n?o transformadas. Na raiz os NRT2 mostraram baixa express?o nas linhagens L1 e L4 quando submetidas ao suprimento constante de N-NO3-, em contrapartida, no tratamento sob ressuprimento com 0,2 mM de N-NO3- ocorreu aumento de express?o dos OsNTR2.1~2.2 e NAR nas duas linhagens transformadas, por?m na raiz a concentra??o de N-NO3- foi oposta a express?o dos NRT2 e NAR, em ambas as situa??es. Nas folhas, a linhagem L4 apresentou alta express?o do transportador OsNRT2.1 com o ressuprimento de 0,2 e 2,0 mM de N-NO3-. Nas plantas submetidas ao suprimento constante de N-NH4+, a L1 apresentou menor express?o dos AMT1 na raiz quando comparada a L4 e a planta n?o transformada. Quando submetida a defici?ncia de N-NH4+, a express?o do OsAMT1.2 aumentou nas ra?zes de todas as plantas. Entretanto, n?o houve correla??o positiva entre a express?o dos transportadores de N e os teores de NO3- e NH4+ nas linhagens transformadas, indicando uma poss?vel altera??o na atividade das enzimas de redu??o e ou assimila??o de N. As plantas transformadas quando submetidas ao ressuprimento com baixos teores de nitrato e am?nio apresentaram melhor resposta dos par?metros Vm?x e KM em rela??o a n?o transformadas. Nas plantas transformadas o ressuprimento com nitrato em baixa concentra??o resultou em maior express?o dos genes dos transportadores OsNTR2.1~2.2 e da prote?na OsNAR2.1 e o tratamento com suprimento constante proporcionou maior ac?mulo de nitrato nestas plantas. Os resultados tanto dos par?metros cin?ticos quanto do ac?mulo de mat?ria fresca sugerem que as plantas transformadas para express?o do OsDof25 apresentaram maior toler?ncia ao estresse nutricional.Submitted by Jorge Silva (jorgelmsilva@ufrrj.br) on 2017-05-26T20:35:35Z No. of bitstreams: 1 2012 - Renata Aparecida Costa Silva.pdf: 1687897 bytes, checksum: 4d1af8e117daa1842ae1766319ddf8d6 (MD5)Made available in DSpace on 2017-05-26T20:35:35Z (GMT). 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dc.title.por.fl_str_mv Efeito da superexpress?o do fator de transcri??o OsDof25 sobre a efici?ncia de absor??o de nitrog?nio em Orysa sativa L.
dc.title.alternative.eng.fl_str_mv Effect of superexpression of the transcription factor OsDof25 on the efficiency of nitrogen uptake in Orysa sativa L.
title Efeito da superexpress?o do fator de transcri??o OsDof25 sobre a efici?ncia de absor??o de nitrog?nio em Orysa sativa L.
spellingShingle Efeito da superexpress?o do fator de transcri??o OsDof25 sobre a efici?ncia de absor??o de nitrog?nio em Orysa sativa L.
Silva, Renata Aparecida Costa
Ammonium transporter
Nitrate transporter
Gene expression
Transportador de am?nio
Transportador de nitrato
Express?o g?nica.
Agronomia
title_short Efeito da superexpress?o do fator de transcri??o OsDof25 sobre a efici?ncia de absor??o de nitrog?nio em Orysa sativa L.
title_full Efeito da superexpress?o do fator de transcri??o OsDof25 sobre a efici?ncia de absor??o de nitrog?nio em Orysa sativa L.
title_fullStr Efeito da superexpress?o do fator de transcri??o OsDof25 sobre a efici?ncia de absor??o de nitrog?nio em Orysa sativa L.
title_full_unstemmed Efeito da superexpress?o do fator de transcri??o OsDof25 sobre a efici?ncia de absor??o de nitrog?nio em Orysa sativa L.
title_sort Efeito da superexpress?o do fator de transcri??o OsDof25 sobre a efici?ncia de absor??o de nitrog?nio em Orysa sativa L.
author Silva, Renata Aparecida Costa
author_facet Silva, Renata Aparecida Costa
author_role author
dc.contributor.advisor1.fl_str_mv Souza, Sonia Regina de
dc.contributor.advisor1ID.fl_str_mv 003.927-487-03
dc.contributor.advisor-co1.fl_str_mv Santos, Leandro Azevedo
dc.contributor.advisor-co1ID.fl_str_mv 983.907.835-68
dc.contributor.referee1.fl_str_mv Souza., Sonia Regina de
dc.contributor.referee2.fl_str_mv Ferraz J?nior, Altamiro Souza de Lima |
dc.contributor.referee3.fl_str_mv Cabral, Luiz Mors
dc.contributor.authorID.fl_str_mv 077.966.316-00
dc.contributor.authorLattes.fl_str_mv http://lattes.cnpq.br/6396561004465081
dc.contributor.author.fl_str_mv Silva, Renata Aparecida Costa
contributor_str_mv Souza, Sonia Regina de
Santos, Leandro Azevedo
Souza., Sonia Regina de
Ferraz J?nior, Altamiro Souza de Lima |
Cabral, Luiz Mors
dc.subject.eng.fl_str_mv Ammonium transporter
Nitrate transporter
Gene expression
topic Ammonium transporter
Nitrate transporter
Gene expression
Transportador de am?nio
Transportador de nitrato
Express?o g?nica.
Agronomia
dc.subject.por.fl_str_mv Transportador de am?nio
Transportador de nitrato
Express?o g?nica.
dc.subject.cnpq.fl_str_mv Agronomia
description Nitrogen is one of the nutrient elements most limiting for plant growth. Thus, increasing plant nitrogen usage efficiency (NUE) is an essential factor for sustainable agriculture, leading to an increased food production with less fertilizer input and less environment impact. The aim of this study was to evaluate the effect of OsDof25 overexpression on N-NO3- and N-NH4+ uptake. In transgenic rice plants, OsDof25 was expressed under control of maize ubiquitin promoter (UBIL:OsDof25:3xHA). Two experiments were conducted: one to evaluate the kinetic parameters Vm?x and KM, and another to analyze the expression level of nitrate (NRT2.1~2.2 and NAR) and ammonium transporters (AMT1.1~1.3), both under high and low NO3- and NH4+ supply. The untransformed plants showed higher growth that transformed lineages. The L1 and L2 showed a lower value of the KM in the resupply treatment of 0.2 mM N-NO3-. In the resupply with 0.2 mM N-NH4 + the L4 showed higher Vmax and L1 lower KM. There were no large variations in uptake kinetics between the transformed and untransformed plants. At the root the NRT2 showed low expression in lineages L1 and L4, when under constant supply of N-NO3-, in contrast, in the treatment under resupply with 0.2 mM N-NO3-was increased expression of OsNTR2.1 ~ 2.2, and NAR in both transformed lineages, but in the root the concentration of NO3- was opposed to the expression of NRT2 and NAR, in both conditions. In the leaves, the line L4 showed high expression of the transporter OsNRT2.1 with the resupply of 0.2 and 2.0 mM N-NO3-. In plants grown under constant supply of N-NH4+, L1 showed lower expression of AMT1 in the root compared to L4 and untransformed plants. When subjected to nitrogen deficiency, there was an increased expression of the OsAMT1.2. However, there was no correlation between N transporter expression levels and NO3- and NH4+ content in the transformed plants, indicating a possible change in enzyme activity and reduction or assimilation of N in these plants. The transformed plants when subjected to resupply with low levels of nitrate and ammonium showed better response parameters Vmax and KM compared to the untransformed. In the plants transformed the resupply with nitrate at low concentration resulted in increasing the gene expression of the transporters (OsNTR2.1 ~ 2.2 and protein OsNAR2.1), and the treatment with constant supply provided greatest nitrate accumulation in these plants. The results of both kinetic parameters and accumulation of fresh matter suggest that plants transformed for the expression of the OsDof25 presented highest tolerance to nutritional stress.
publishDate 2012
dc.date.issued.fl_str_mv 2012-02-15
dc.date.accessioned.fl_str_mv 2017-05-26T20:35:35Z
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 SILVA, Renata Aparecida Costa. Efeito da superexpress?o do fator de transcri??o OsDof25 sobre a efici?ncia de absor??o de nitrog?nio em Orysa sativa L. 2012. 52 f. Disserta??o (Mestrado em Agronomia - Ci?ncia do Solo). Instituto de Agronomia, Departamento de Solos, Universidade Federal Rural do Rio de Janeiro, Serop?dica, 2012.
dc.identifier.uri.fl_str_mv https://tede.ufrrj.br/jspui/handle/jspui/1692
identifier_str_mv SILVA, Renata Aparecida Costa. Efeito da superexpress?o do fator de transcri??o OsDof25 sobre a efici?ncia de absor??o de nitrog?nio em Orysa sativa L. 2012. 52 f. Disserta??o (Mestrado em Agronomia - Ci?ncia do Solo). Instituto de Agronomia, Departamento de Solos, Universidade Federal Rural do Rio de Janeiro, Serop?dica, 2012.
url https://tede.ufrrj.br/jspui/handle/jspui/1692
dc.language.iso.fl_str_mv por
language por
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
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dc.publisher.none.fl_str_mv Universidade Federal Rural do Rio de Janeiro
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