Defesa antioxidativa em plantas de arroz duplamente silenciadas nas APXs citosólicas e expostas a estresses abióticos

Detalhes bibliográficos
Autor(a) principal: Fontenele, Adilton de Vasconcelos
Data de Publicação: 2011
Tipo de documento: Dissertação
Idioma: por
Título da fonte: Repositório Institucional da Universidade Federal do Ceará (UFC)
Texto Completo: http://www.repositorio.ufc.br/handle/riufc/4830
Resumo: The aim of this study was to characterize physiological and biochemical aspects that show if rice plants (Oryza sativa) knockdown on cytosolic ascorbate peroxidase enzyme (cAPX) are more susceptible to oxidative stress than the wild type plants. APX is an important enzyme from oxidative metabolism of plants, acting on regulation of the endogenous levels of hydrogen peroxide (H2O2). For this, rice plants knockdown on cAPX (Apx1/2s) and wild type (WT) were used for the experimentation. The plants were silenced by interference RNA technical (iRNA) and were grown for 35 days into 1.5 L pots containing nutritive solution under greenhouse conditions. The experiment I was performed with leaves segments immersed in methyl viologen (MV) 50 μM for 24h and the experiment II was performed by application of the following treatments: salt stress, high light and MV. The results from experiment I shown Apx1/2s plants have a higher level of H2O2 high than the levels found on WT rice plants, suggesting that Apx1/2s plants present a level of H2O2 near a level of cell signaling. The fact of WT plants had accumulated H2O2 1h after the treatment suggest the necessity of a signaling H2O2 level for stimulate defense systems. Apx1/2s plants unlike of WT plants presented a constant decline on H2O2 content, indicating a likely H2O2 scavenging excess. After 3h of treatment the chloroplastic enzymes SOD, APX and PHGPx presented upper active in Apx1/2s plants, in control and stress, compared with WT plants. These results suggest the existence of an antioxidant system quite active in the Apx1/2s plants. In experiment II the Apx1/2s plants presented no differences in the photochemical parameters when compared with WT plants, even possessing a smaller photosynthesis under controlled conditions. The energy dissipation (NPQ) in the Apx1/2s plants under high light was, in average, higher than WT plants, suggesting better energy dissipation. Even with efficient energy dissipation, the plants could not avoid the excess of energy in the photosystem and they suffered photoinhibition and damage in photosynthetic apparatus (Fv/Fm). In relation the WT plants, Apx1/2s plants presented a higher activity of antioxidant enzymes SOD, CAT and PHGPx under controlled conditions, probably intending compensate the lack of cAPX. In the MV treatment the chloroplastic PHGPx was stimulated above 100% in the Apx1/2s plants, indicating that these plants can repair oxidative damage faster than the WT plants. The results suggest that Apx1/2s plants, despite the absence of cAPX, activated additional security systems to compensate the lack of cAPX and respond quickly and efficiently to stress situations.
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spelling Defesa antioxidativa em plantas de arroz duplamente silenciadas nas APXs citosólicas e expostas a estresses abióticosBioquímicaEstresse oxidativoFluorescência da clorofilaPeroxidase do ascorbatoThe aim of this study was to characterize physiological and biochemical aspects that show if rice plants (Oryza sativa) knockdown on cytosolic ascorbate peroxidase enzyme (cAPX) are more susceptible to oxidative stress than the wild type plants. APX is an important enzyme from oxidative metabolism of plants, acting on regulation of the endogenous levels of hydrogen peroxide (H2O2). For this, rice plants knockdown on cAPX (Apx1/2s) and wild type (WT) were used for the experimentation. The plants were silenced by interference RNA technical (iRNA) and were grown for 35 days into 1.5 L pots containing nutritive solution under greenhouse conditions. The experiment I was performed with leaves segments immersed in methyl viologen (MV) 50 μM for 24h and the experiment II was performed by application of the following treatments: salt stress, high light and MV. The results from experiment I shown Apx1/2s plants have a higher level of H2O2 high than the levels found on WT rice plants, suggesting that Apx1/2s plants present a level of H2O2 near a level of cell signaling. The fact of WT plants had accumulated H2O2 1h after the treatment suggest the necessity of a signaling H2O2 level for stimulate defense systems. Apx1/2s plants unlike of WT plants presented a constant decline on H2O2 content, indicating a likely H2O2 scavenging excess. After 3h of treatment the chloroplastic enzymes SOD, APX and PHGPx presented upper active in Apx1/2s plants, in control and stress, compared with WT plants. These results suggest the existence of an antioxidant system quite active in the Apx1/2s plants. In experiment II the Apx1/2s plants presented no differences in the photochemical parameters when compared with WT plants, even possessing a smaller photosynthesis under controlled conditions. The energy dissipation (NPQ) in the Apx1/2s plants under high light was, in average, higher than WT plants, suggesting better energy dissipation. Even with efficient energy dissipation, the plants could not avoid the excess of energy in the photosystem and they suffered photoinhibition and damage in photosynthetic apparatus (Fv/Fm). In relation the WT plants, Apx1/2s plants presented a higher activity of antioxidant enzymes SOD, CAT and PHGPx under controlled conditions, probably intending compensate the lack of cAPX. In the MV treatment the chloroplastic PHGPx was stimulated above 100% in the Apx1/2s plants, indicating that these plants can repair oxidative damage faster than the WT plants. The results suggest that Apx1/2s plants, despite the absence of cAPX, activated additional security systems to compensate the lack of cAPX and respond quickly and efficiently to stress situations.O objetivo do presente trabalho foi caracterizar aspectos fisiológicos e bioquímicos que mostrem se plantas de arroz (Oryza sativa) deficientes da enzima citosólica peroxidase do ascorbato (cAPX) são mais suscetíveis ao estresse oxidativo do que as plantas com cAPX. A APX é uma importante enzima do metabolismo oxidativo de plantas, atuando na regulação dos níveis endógenos do peróxido de hidrogênio (H2O2). Para isso, plantas deficientes em cAPX (Apx1/2s) e plantas não transformadas (WT) foram utilizadas para a experimentação. As plantas foram silenciadas pela técnica do RNA de interferência (iRNA) e cultivadas por 35 dias em vasos de 1.5 L contendo solução nutritiva sob condições de casa de vegetação. O experimento I foi realizado com segmentos de folhas imersos em metil-viologênio (MV) 50 M durante 24h, e o experimento II foi realizado pela aplicação dos seguintes estresses abióticos: salinidade, alta luminosidade e MV. Os resultados do experimento I mostraram que as plantas Apx1/2s possuem um nível basal de H2O2 maior do que os níveis encontrados nas plantas WT, sugerindo que as plantas Apx1/2s apresentam um nível de H2O2 próximo de um nível de sinalização celular. O fato das plantas WT terem acumulado H2O2 após 1h de tratamento sugere a necessidade de um nível sinalizador de H2O2 para ativação dos sistemas de defesa. As plantas Apx1/2s ao contrario das WT apresentaram uma queda constante no conteúdo de H2O2, indicando uma provável remoção do excesso de H2O2. Após 3h de tratamento as enzimas SOD, APX e PHGPx de cloroplasto apresentaram atividade superior nas plantas Apx1/2s, tanto no controle quanto no estresse, comparadas com as plantas WT. Esses resultados sugerem existência de um sistema antioxidante bastante ativado nas plantas Apx1/2s. No experimento II as plantas Apx1/2s não apresentaram diferenças nos parâmetros fotoquímicos quando comparadas com as plantas WT, mesmo possuindo uma menor fotossíntese em condições controle. A dissipação do excesso de energia (NPQ) nas plantas Apx1/2s tratadas com luz foi, em média, maior que das plantas WT, indicando uma possível maior eficiência na dissipação de energia. Mesmo com eficiente dissipação de energia ambas as plantas não conseguiram evitar energia excessiva no fotossistema e acabaram sofrendo fotoinibição e danos no aparato fotossintético (Fv/Fm). Em relação às plantas WT, as Apx1/2s apresentaram maior atividade das enzimas antioxidativas SOD, CAT e PHGPx nas condições controle, na provável tentativa de compensar a ausência da cAPX. No tratamento com MV a isoforma cloroplástica da PHGPx foi estimulada em mais de 100% nas plantas Apx1/2s, indicando que essas plantas podem reparar danos oxidativos com mais rapidez que as WT. Os dados sugerem que as plantas Apx1/2s, apesar da ausência da cAPX, ativam sistemas adicionais de proteção antioxidativa para compensar essa ausência e responder mais rápida e eficientemente a situações de estresse.Silveira, Joaquim Albenísio Gomes daFontenele, Adilton de Vasconcelos2013-05-20T19:17:35Z2013-05-20T19:17:35Z2011info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisapplication/pdfFONTENELE, A. V. de. Defesa antioxidativa em plantas de arroz duplamente silenciadas nas APXs citosólicas e expostas a estresses abióticos. 93 f. 2011. Dissertação (Mestrado em Bioquímica) - Centro de Ciências, Universidade Federal do Ceará, Fortaleza, 2011.http://www.repositorio.ufc.br/handle/riufc/4830porreponame:Repositório Institucional da Universidade Federal do Ceará (UFC)instname:Universidade Federal do Ceará (UFC)instacron:UFCinfo:eu-repo/semantics/openAccess2020-05-25T16:28:13Zoai:repositorio.ufc.br:riufc/4830Repositório InstitucionalPUBhttp://www.repositorio.ufc.br/ri-oai/requestbu@ufc.br || repositorio@ufc.bropendoar:2020-05-25T16:28:13Repositório Institucional da Universidade Federal do Ceará (UFC) - Universidade Federal do Ceará (UFC)false
dc.title.none.fl_str_mv Defesa antioxidativa em plantas de arroz duplamente silenciadas nas APXs citosólicas e expostas a estresses abióticos
title Defesa antioxidativa em plantas de arroz duplamente silenciadas nas APXs citosólicas e expostas a estresses abióticos
spellingShingle Defesa antioxidativa em plantas de arroz duplamente silenciadas nas APXs citosólicas e expostas a estresses abióticos
Fontenele, Adilton de Vasconcelos
Bioquímica
Estresse oxidativo
Fluorescência da clorofila
Peroxidase do ascorbato
title_short Defesa antioxidativa em plantas de arroz duplamente silenciadas nas APXs citosólicas e expostas a estresses abióticos
title_full Defesa antioxidativa em plantas de arroz duplamente silenciadas nas APXs citosólicas e expostas a estresses abióticos
title_fullStr Defesa antioxidativa em plantas de arroz duplamente silenciadas nas APXs citosólicas e expostas a estresses abióticos
title_full_unstemmed Defesa antioxidativa em plantas de arroz duplamente silenciadas nas APXs citosólicas e expostas a estresses abióticos
title_sort Defesa antioxidativa em plantas de arroz duplamente silenciadas nas APXs citosólicas e expostas a estresses abióticos
author Fontenele, Adilton de Vasconcelos
author_facet Fontenele, Adilton de Vasconcelos
author_role author
dc.contributor.none.fl_str_mv Silveira, Joaquim Albenísio Gomes da
dc.contributor.author.fl_str_mv Fontenele, Adilton de Vasconcelos
dc.subject.por.fl_str_mv Bioquímica
Estresse oxidativo
Fluorescência da clorofila
Peroxidase do ascorbato
topic Bioquímica
Estresse oxidativo
Fluorescência da clorofila
Peroxidase do ascorbato
description The aim of this study was to characterize physiological and biochemical aspects that show if rice plants (Oryza sativa) knockdown on cytosolic ascorbate peroxidase enzyme (cAPX) are more susceptible to oxidative stress than the wild type plants. APX is an important enzyme from oxidative metabolism of plants, acting on regulation of the endogenous levels of hydrogen peroxide (H2O2). For this, rice plants knockdown on cAPX (Apx1/2s) and wild type (WT) were used for the experimentation. The plants were silenced by interference RNA technical (iRNA) and were grown for 35 days into 1.5 L pots containing nutritive solution under greenhouse conditions. The experiment I was performed with leaves segments immersed in methyl viologen (MV) 50 μM for 24h and the experiment II was performed by application of the following treatments: salt stress, high light and MV. The results from experiment I shown Apx1/2s plants have a higher level of H2O2 high than the levels found on WT rice plants, suggesting that Apx1/2s plants present a level of H2O2 near a level of cell signaling. The fact of WT plants had accumulated H2O2 1h after the treatment suggest the necessity of a signaling H2O2 level for stimulate defense systems. Apx1/2s plants unlike of WT plants presented a constant decline on H2O2 content, indicating a likely H2O2 scavenging excess. After 3h of treatment the chloroplastic enzymes SOD, APX and PHGPx presented upper active in Apx1/2s plants, in control and stress, compared with WT plants. These results suggest the existence of an antioxidant system quite active in the Apx1/2s plants. In experiment II the Apx1/2s plants presented no differences in the photochemical parameters when compared with WT plants, even possessing a smaller photosynthesis under controlled conditions. The energy dissipation (NPQ) in the Apx1/2s plants under high light was, in average, higher than WT plants, suggesting better energy dissipation. Even with efficient energy dissipation, the plants could not avoid the excess of energy in the photosystem and they suffered photoinhibition and damage in photosynthetic apparatus (Fv/Fm). In relation the WT plants, Apx1/2s plants presented a higher activity of antioxidant enzymes SOD, CAT and PHGPx under controlled conditions, probably intending compensate the lack of cAPX. In the MV treatment the chloroplastic PHGPx was stimulated above 100% in the Apx1/2s plants, indicating that these plants can repair oxidative damage faster than the WT plants. The results suggest that Apx1/2s plants, despite the absence of cAPX, activated additional security systems to compensate the lack of cAPX and respond quickly and efficiently to stress situations.
publishDate 2011
dc.date.none.fl_str_mv 2011
2013-05-20T19:17:35Z
2013-05-20T19:17:35Z
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.uri.fl_str_mv FONTENELE, A. V. de. Defesa antioxidativa em plantas de arroz duplamente silenciadas nas APXs citosólicas e expostas a estresses abióticos. 93 f. 2011. Dissertação (Mestrado em Bioquímica) - Centro de Ciências, Universidade Federal do Ceará, Fortaleza, 2011.
http://www.repositorio.ufc.br/handle/riufc/4830
identifier_str_mv FONTENELE, A. V. de. Defesa antioxidativa em plantas de arroz duplamente silenciadas nas APXs citosólicas e expostas a estresses abióticos. 93 f. 2011. Dissertação (Mestrado em Bioquímica) - Centro de Ciências, Universidade Federal do Ceará, Fortaleza, 2011.
url http://www.repositorio.ufc.br/handle/riufc/4830
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
dc.format.none.fl_str_mv application/pdf
dc.source.none.fl_str_mv reponame:Repositório Institucional da Universidade Federal do Ceará (UFC)
instname:Universidade Federal do Ceará (UFC)
instacron:UFC
instname_str Universidade Federal do Ceará (UFC)
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institution UFC
reponame_str Repositório Institucional da Universidade Federal do Ceará (UFC)
collection Repositório Institucional da Universidade Federal do Ceará (UFC)
repository.name.fl_str_mv Repositório Institucional da Universidade Federal do Ceará (UFC) - Universidade Federal do Ceará (UFC)
repository.mail.fl_str_mv bu@ufc.br || repositorio@ufc.br
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