Establishment of a 13C-positional isotope labelling approach and comparative investigation of metabolic fluxes in mesophyll and guard cells
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Data de Publicação: | 2020 |
Tipo de documento: | Dissertação |
Idioma: | eng |
Título da fonte: | Repositório Institucional da UFLA |
Texto Completo: | http://repositorio.ufla.br/jspui/handle/1/43232 |
Resumo: | Guard cells respond to a wide range of stimuli in order to change its volume and, thus, open or close the stomatal pore. Several stimuli cause changes in guard cells turgor, such as light, CO2, phytohormones, among others. Guard cells metabolism have differences in comparison with mesophyll cells. For instance, whilst the metabolic flux throughout the tricarboxylic acid (TCA) cycle is inhibited by light in mesophyll cells, evidences suggest that guard cells TCA cycle is active in presence of light. In this sense, this work aimed to investigate the metabolic fluxes through the TCA cycle and other associated pathways, such as glycolysis and CO2 fixation mediated by phosphoenolpyruvate carboxylase (PEPc). For this purpose, we stablished a mass spectrometry-based 13C-positional isotope labelling approach to analyse previously published gas-chromatography coupled to mass spectrometry data. Several metabolites of, or associated to the TCA cycle were evaluated in terms of relative isotopomer abundance, total 13C-enrichment and metabolite content, and the results from guard cells were compared with mesophyll cells of C3 or C4 plants, as well as to source or sink C3-mesophyll cells. In this sense, the results suggest that guard cells have a particular metabolic flux mode throughout TCA cycle and associated pathways, with certain similarities with mesophyll cells of C3 plants. Furthermore, the results indicate that the light-dependent inhibition of pyruvate dehydrogenase severely restricts the fluxes of carbon to glutamate synthesis. Results are discussed in the context of TCA cycle regulation and guard cell metabolism. |
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Establishment of a 13C-positional isotope labelling approach and comparative investigation of metabolic fluxes in mesophyll and guard cellsEstabelecimento de abordagem posicional isotópica de 13 C e investigação comparativa de fluxos metabólicos em células mesofílicas e células-guardaCiclo do ácido tricarboxílicoFluxo metabólicoGlutaminaMarcação isotópica posicionalMetabolismo de células guardaRegulação metabólica13C-positional isotope labellingGlutamineGuard cells metabolismMetabolic fluxesMetabolic regulationTricarboxylic acid cycleFisiologia VegetalGuard cells respond to a wide range of stimuli in order to change its volume and, thus, open or close the stomatal pore. Several stimuli cause changes in guard cells turgor, such as light, CO2, phytohormones, among others. Guard cells metabolism have differences in comparison with mesophyll cells. For instance, whilst the metabolic flux throughout the tricarboxylic acid (TCA) cycle is inhibited by light in mesophyll cells, evidences suggest that guard cells TCA cycle is active in presence of light. In this sense, this work aimed to investigate the metabolic fluxes through the TCA cycle and other associated pathways, such as glycolysis and CO2 fixation mediated by phosphoenolpyruvate carboxylase (PEPc). For this purpose, we stablished a mass spectrometry-based 13C-positional isotope labelling approach to analyse previously published gas-chromatography coupled to mass spectrometry data. Several metabolites of, or associated to the TCA cycle were evaluated in terms of relative isotopomer abundance, total 13C-enrichment and metabolite content, and the results from guard cells were compared with mesophyll cells of C3 or C4 plants, as well as to source or sink C3-mesophyll cells. In this sense, the results suggest that guard cells have a particular metabolic flux mode throughout TCA cycle and associated pathways, with certain similarities with mesophyll cells of C3 plants. Furthermore, the results indicate that the light-dependent inhibition of pyruvate dehydrogenase severely restricts the fluxes of carbon to glutamate synthesis. Results are discussed in the context of TCA cycle regulation and guard cell metabolism.Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)As células-guarda respondem à diversos estímulos que resultam na alteração do seu volume e, como consequência, na abertura ou fechamento do poro estomático. Diversos estímulos ocasionam mudanças no volume das células guarda, como por exemplo luz, CO2 e fitohormônios. As células-guarda possuem um metabolismo com diversas particularidades, em comparação com as células mesofílicas. Por exemplo, enquanto o fluxo metabólico através do ciclo do ácido tricarboxílico (TCA) é inibido pela luz em células mesofílicas, evidências sugerem que o ciclo TCA e a glicólise de células guarda sejam ativados na presença de luz. Dentro desta perspectiva, este trabalho objetivou investigar os fluxos metabólicos através do ciclo TCA e de rotas metabólicas associadas, como da glicólise, da fixação de CO2 mediada pela enzima fosfoenolpiruvatocarboxilase (PEPc) e da síntese de amino ácidos como Ala, Glu e Gln. Para isso, foi estabelecido uma abordagem de marcação isotópica (13C) posicional utilizando dados previamente publicados de cromatografia gasosa acoplada à espectrometria de massas. Diversos metabólitos do ciclo TCA e de rotas metabólicas associadas foram avaliados em termos de abundância relativa do isotopômero, enriquecimento total de 13C e conteúdo dos metabólitos. Os resultados obtidos para células-guarda foram comparados com resultados de células mesofílicas de plantas C3 e C4, bem como com os de células mesofílicas de folhas fonte e dreno de plantas C3. Nesse sentido, os resultados sugerem que as células-guarda têm um modo particular de fluxo do ciclo TCA, e das rotas associadas, com certas similaridades com células-dreno de plantas C3. Além disso, os resultados indicam que a inibição dependente de luz da piruvato desidrogenase restringe severamente o fluxo do ciclo TCA em direção à síntese de glutamato.Os resultados são discutidos no contexto da regulação do ciclo TCA e do metabolismo de células guarda.Universidade Federal de LavrasPrograma de Pós-Graduação em Agronomia/Fisiologia VegetalUFLAbrasilDepartamento de BiologiaAnjos, Letícia dosDaloso, Danilo de MenezesAnjos, Letícia dosDaloso, Danilo de MenezesMedeiros, David BarbosaGarcia, Rebeca Patrícia OmenaReis, André Guilherme Daubermann2020-09-29T12:22:50Z2020-09-29T12:22:50Z2020-09-292020-08-10info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisapplication/pdfREIS, A. G. D. Establishment of a 13C-positional isotope labelling approach and comparative investigation of metabolic fluxes in mesophyll and guard cells. 2020. 76 p. Dissertação (Mestrado em Agronomia/Fisiologia Vegetal) – Universidade Federal de Lavras, Lavras, 2020.http://repositorio.ufla.br/jspui/handle/1/43232enginfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da UFLAinstname:Universidade Federal de Lavras (UFLA)instacron:UFLA2023-04-27T16:18:32Zoai:localhost:1/43232Repositório InstitucionalPUBhttp://repositorio.ufla.br/oai/requestnivaldo@ufla.br || repositorio.biblioteca@ufla.bropendoar:2023-04-27T16:18:32Repositório Institucional da UFLA - Universidade Federal de Lavras (UFLA)false |
dc.title.none.fl_str_mv |
Establishment of a 13C-positional isotope labelling approach and comparative investigation of metabolic fluxes in mesophyll and guard cells Estabelecimento de abordagem posicional isotópica de 13 C e investigação comparativa de fluxos metabólicos em células mesofílicas e células-guarda |
title |
Establishment of a 13C-positional isotope labelling approach and comparative investigation of metabolic fluxes in mesophyll and guard cells |
spellingShingle |
Establishment of a 13C-positional isotope labelling approach and comparative investigation of metabolic fluxes in mesophyll and guard cells Reis, André Guilherme Daubermann Ciclo do ácido tricarboxílico Fluxo metabólico Glutamina Marcação isotópica posicional Metabolismo de células guarda Regulação metabólica 13C-positional isotope labelling Glutamine Guard cells metabolism Metabolic fluxes Metabolic regulation Tricarboxylic acid cycle Fisiologia Vegetal |
title_short |
Establishment of a 13C-positional isotope labelling approach and comparative investigation of metabolic fluxes in mesophyll and guard cells |
title_full |
Establishment of a 13C-positional isotope labelling approach and comparative investigation of metabolic fluxes in mesophyll and guard cells |
title_fullStr |
Establishment of a 13C-positional isotope labelling approach and comparative investigation of metabolic fluxes in mesophyll and guard cells |
title_full_unstemmed |
Establishment of a 13C-positional isotope labelling approach and comparative investigation of metabolic fluxes in mesophyll and guard cells |
title_sort |
Establishment of a 13C-positional isotope labelling approach and comparative investigation of metabolic fluxes in mesophyll and guard cells |
author |
Reis, André Guilherme Daubermann |
author_facet |
Reis, André Guilherme Daubermann |
author_role |
author |
dc.contributor.none.fl_str_mv |
Anjos, Letícia dos Daloso, Danilo de Menezes Anjos, Letícia dos Daloso, Danilo de Menezes Medeiros, David Barbosa Garcia, Rebeca Patrícia Omena |
dc.contributor.author.fl_str_mv |
Reis, André Guilherme Daubermann |
dc.subject.por.fl_str_mv |
Ciclo do ácido tricarboxílico Fluxo metabólico Glutamina Marcação isotópica posicional Metabolismo de células guarda Regulação metabólica 13C-positional isotope labelling Glutamine Guard cells metabolism Metabolic fluxes Metabolic regulation Tricarboxylic acid cycle Fisiologia Vegetal |
topic |
Ciclo do ácido tricarboxílico Fluxo metabólico Glutamina Marcação isotópica posicional Metabolismo de células guarda Regulação metabólica 13C-positional isotope labelling Glutamine Guard cells metabolism Metabolic fluxes Metabolic regulation Tricarboxylic acid cycle Fisiologia Vegetal |
description |
Guard cells respond to a wide range of stimuli in order to change its volume and, thus, open or close the stomatal pore. Several stimuli cause changes in guard cells turgor, such as light, CO2, phytohormones, among others. Guard cells metabolism have differences in comparison with mesophyll cells. For instance, whilst the metabolic flux throughout the tricarboxylic acid (TCA) cycle is inhibited by light in mesophyll cells, evidences suggest that guard cells TCA cycle is active in presence of light. In this sense, this work aimed to investigate the metabolic fluxes through the TCA cycle and other associated pathways, such as glycolysis and CO2 fixation mediated by phosphoenolpyruvate carboxylase (PEPc). For this purpose, we stablished a mass spectrometry-based 13C-positional isotope labelling approach to analyse previously published gas-chromatography coupled to mass spectrometry data. Several metabolites of, or associated to the TCA cycle were evaluated in terms of relative isotopomer abundance, total 13C-enrichment and metabolite content, and the results from guard cells were compared with mesophyll cells of C3 or C4 plants, as well as to source or sink C3-mesophyll cells. In this sense, the results suggest that guard cells have a particular metabolic flux mode throughout TCA cycle and associated pathways, with certain similarities with mesophyll cells of C3 plants. Furthermore, the results indicate that the light-dependent inhibition of pyruvate dehydrogenase severely restricts the fluxes of carbon to glutamate synthesis. Results are discussed in the context of TCA cycle regulation and guard cell metabolism. |
publishDate |
2020 |
dc.date.none.fl_str_mv |
2020-09-29T12:22:50Z 2020-09-29T12:22:50Z 2020-09-29 2020-08-10 |
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 |
REIS, A. G. D. Establishment of a 13C-positional isotope labelling approach and comparative investigation of metabolic fluxes in mesophyll and guard cells. 2020. 76 p. Dissertação (Mestrado em Agronomia/Fisiologia Vegetal) – Universidade Federal de Lavras, Lavras, 2020. http://repositorio.ufla.br/jspui/handle/1/43232 |
identifier_str_mv |
REIS, A. G. D. Establishment of a 13C-positional isotope labelling approach and comparative investigation of metabolic fluxes in mesophyll and guard cells. 2020. 76 p. Dissertação (Mestrado em Agronomia/Fisiologia Vegetal) – Universidade Federal de Lavras, Lavras, 2020. |
url |
http://repositorio.ufla.br/jspui/handle/1/43232 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
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.publisher.none.fl_str_mv |
Universidade Federal de Lavras Programa de Pós-Graduação em Agronomia/Fisiologia Vegetal UFLA brasil Departamento de Biologia |
publisher.none.fl_str_mv |
Universidade Federal de Lavras Programa de Pós-Graduação em Agronomia/Fisiologia Vegetal UFLA brasil Departamento de Biologia |
dc.source.none.fl_str_mv |
reponame:Repositório Institucional da UFLA instname:Universidade Federal de Lavras (UFLA) instacron:UFLA |
instname_str |
Universidade Federal de Lavras (UFLA) |
instacron_str |
UFLA |
institution |
UFLA |
reponame_str |
Repositório Institucional da UFLA |
collection |
Repositório Institucional da UFLA |
repository.name.fl_str_mv |
Repositório Institucional da UFLA - Universidade Federal de Lavras (UFLA) |
repository.mail.fl_str_mv |
nivaldo@ufla.br || repositorio.biblioteca@ufla.br |
_version_ |
1807835140516216832 |