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  • About
  • The Global ETD Search service is a free service for researchers to find electronic theses and dissertations. This service is provided by the Networked Digital Library of Theses and Dissertations.
    Our metadata is collected from universities around the world. If you manage a university/consortium/country archive and want to be added, details can be found on the NDLTD website.
1

Estudo do metabolismo dos lipÃdeos de membranas do cloroplasto e dos genes associados em Vigna unguiculata (L.) Walp. em condiÃÃo de dÃficit hÃdrico e reidrataÃÃo subseqÃente / Study of chloroplast membrane lipids metabolism and the associated genes in Vigna unguiculata (L.) Walp. under drought and recovery after rehydration

Maria Lucia Torres Franklin 19 December 2008 (has links)
Conselho Nacional de Desenvolvimento CientÃfico e TecnolÃgico / FundaÃÃo Cearense de Apoio ao Desenvolvimento Cientifico e TecnolÃgico / As membranas biolÃgicas sÃo alvos preferenciais dos efeitos deletÃrios do estresse hÃdrico, induzidos por aÃÃo de enzimas hidrolÃticas e espÃcies reativas do oxigÃnio (ERO), ambas estimuladas durante o estresse. A biossÃntese dos lipÃdeos dos cloroplastos pode ser importante para a tolerÃncia ao estresse hÃdrico e para a recuperaÃÃo apÃs reidrataÃÃo. Nesse trabalho nos estudamos o metabolismo dos cloroplastos, monogalactosil-diacilglicerol (MGDG), digalactosil-diacilglicerol (DGDG), sulfoquinovosil-diacilglicerol (SQDG), phosphatidil-glicerol (PG), no Ãmbito do dÃficit hÃdrico e da reidrataÃÃo apÃs o fim do estresse. Com este intuito, nos medimos o teor dos lipÃdeos da folhas, acompanhamos a incorporaÃÃo do precursor 14C-acetato nos lipÃdeos e analisamos a expressÃo dos genes codificadores das enzimas de sÃntese chave dos lipÃdeos (MGD1, MGD2, DGD1, DGD2, SQD2 e PGP1) durante o estresse hÃdrico e apÃs a reidrataÃÃo. Visando de uma melhor compreensÃo da relaÃÃo entre o metabolismo destes lipÃdeos e a tolerÃncia a seca, nos trabalhamos com duas cultivares de Vigna unguiculata L. Walp, uma tolerante (cv. EPACE) e outra sensÃvel (cv. 1183) a seca. Por meio de varredura diferencial de um biblioteca de cDNA de V.unguiculata, foram obtidas as seqÃÃncias completas dos cDNA dos genes VuMGD1, VuMGD2, VuDGD1, VuDGD2, VuSQD2 e VuPGP1. Os resultados mostram que em condiÃÃes de estresse hÃdrico a cultivar tolerante, alÃm de preservar seu teor de lipÃdeos durante a seca, à igualmente capaz de estimular a biossÃntese do DGDG aumentando significativamente a relaÃÃo DGDG:MGDG de suas membranas. NÃs sugerimos que o DGDG acumulado em condiÃÃo de seca à transportado para as membranas externas ao cloroplasto e que isso contribui para a tolerÃncia à seca. Os efeitos da desidrataÃÃo celular sobre as membranas tÃm conseqÃÃncias diretas sobre a capacidade das plantas a se recuperarem apÃs reidrataÃÃo. 48 horas apÃs a rega, a cv. sensÃvel 1183 nÃo à capaz de se recuperar em termos de teor de galactolipÃdeos e incorporaÃÃo do precursor. Na cv. tolerante, no entanto, o teor de DGDG permanece elevado, mesmo apÃs a reidrataÃÃo. Em conclusÃo, nossos resultados sugerem a importÃncia dos lipÃdeos membranares na tolerÃncia/sensibilidade das plantas ao dÃficit hÃdrico, em particular o balanÃo entre as classes lipÃdicas de propriedades fÃsico-quÃmicas diferentes (SQDG versus PG e DGDG versus MGDG) que poderiam afetar a estrutura e o funcionamento das membranas. / Membranes are main targets of degradation by reactive oxygen species and hydrolytic activities induced by drought. Chloroplasts lipid biosynthesis, especially galactolipids monogalactosyl-diacylglycerol (MGDG) and digalactosyl-diacylglycerol (DGDG) are important for plant tolerance to water deficit and for recovery after rehydration. In this thesis, we studied the metabolism of the chloroplast membrane lipids, MGDG, DGDG, sulphoquinovosyl-diacylglycerol (SQDG), phosphatidyl-glycerol (PG) under drought and during recovery from drought. Aiming this, we measured leaf lipids content, followed 14C-acÃtate incorporation and expression of genes coding for chloroplast membrane lipid synthases (MGD1, MGD2, DGD1, DGD2, SQD2 and PGP1) during drought and recovery. In order to better understand the relationship between drought tolerance and lipid metabolism, two cultivars of Vigna unguiculata L. Walp, one drought tolerant (cv. EPACE) the other drought susceptible (cv. 1183) were compared. The cDNA complete sequences for VuMGD1, VuMGD2, VuDGD1, VuDGD2, VuSQD2 and VuPGP1 were obtained from screening of a V.unguiculata cDNA library. The results showed that under water stress conditions, the tolerant cultivar, besides its ability to preserve its lipids pool despites drought, is able to strongly stimulate the DGDG biosynthesis, increasing the DGDG:MGDG ratio in its membranes. We suggest that DGDG accumulated under drought condition, when phosphate is deficient, is exported for extrachloroplastic membranes, and thus contributes to plant drought tolerance. Effects of loss of water on cell membranes have direct consequences on plant capacity to recover from stress. 48 hours after rewatering, the susceptible cv. 1183 was not able to fully recover from a moderate stress in terms of leaf galactolipid content and acetate incorporation into MGDG. In EPACE-1, MGDG leaf content remained unchanged after rehydration and DGDG remained higher than in the control plants. In conclusion, our results highlight the importance of membrane lipids in plant adaptation to water deficit and in their capacity to recover from stress. Of particular importance is the balance between lipid classes with various physico-chemical properties (SQDG versus PG, DGDG versus MGDG), since they most likely have a profound influence on membrane structure and function.

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