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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

AnÃlise fisiolÃgica, bioquÃmica e proteÃmica de respostas ao estresse salino em plantas de feijÃo de corda [Vigna unguiculata (L.) Walp.] / Physiologic, biochemistry and proteomic analysis of responses to salt stress in plants of cowpea [Vigna unguiculata (L.) Walp.]

Carlos Eduardo Braga de Abreu 28 September 2012 (has links)
Conselho Nacional de Desenvolvimento CientÃfico e TecnolÃgico / No presente trabalho foi realizado um estudo integrado de fisiologia, bioquÃmica e proteÃmica comparativa em feijÃo de corda, uma cultura de grande valor econÃmico, com o objetivo de entender respostas de aclimataÃÃo das plantas à salinidade. Para tanto, dois experimentos foram conduzidos em casa de vegetaÃÃo, sob condiÃÃes hidropÃnicas, utilizando dois cultivares de feijÃo de corda (Vigna unguiculata) com tolerÃncia diferencial ao estresse salino: PitiÃba (tolerante) e TVu 2331 (sensÃvel). No primeiro experimento foram avaliadas as mudanÃas fisiolÃgicas (crescimento, trocas gasosas, teor relativo de Ãgua, teor de clorofila, fluorescÃncia da clorofila) e bioquÃmicas (acÃmulo de Ãons e solutos orgÃnicos em folhas e raÃzes e padrÃo de expressÃo protÃico foliar) induzidas por concentraÃÃes crescentes de salinidade (NaCl a 50, 75 e 100 mM). Os resultados demonstraram a existÃncia de respostas contrastantes entre os cultivares estudados, especialmente com relaÃÃo à inibiÃÃo do crescimento da parte aÃrea e ao acÃmulo de solutos compatÃveis, os quais foram maiores no TVu. Contudo, a salinidade nÃo alterou os parÃmetros de fluorescÃncia da clorofila em ambos os cultivares. ConcentraÃÃes crescentes de NaCl alteraram de forma diferencial o padrÃo de expressÃo de proteÃnas nas folhas, com maiores alteraÃÃes na dose 100 mM de NaCl. Apesar disso, no segundo experimento, a concentraÃÃo moderada de sal com NaCl a 75 mM foi escolhida como referÃncia para o estudo das mudanÃas no proteoma durante o estresse salino e apÃs um perÃodo pÃs-estresse (recuperaÃÃo). A salinidade modificou a expressÃo de 22 âspotsâ protÃicos no PitiÃba, dos quais 10 (6 que aumentaram e 4 que diminuÃram) tiveram suas identidades determinadas por espectrometria de massas acoplada a cromatografia lÃquida (LC-ESI-MS/MS). No TVu foram observadas mudanÃas em 27 âspotsâ, sendo determinada a identidade de 9 (5 que aumentaram e 4 que diminuÃram). AlÃm destes, 5 âspotsâ que mantiveram suas taxas de expressÃes em condiÃÃes de salinidade no PitiÃba tambÃm foram identificados. A maioria das proteÃnas identificadas està relacionada com o processo de fotossÃntese, realizando funÃÃes enzimÃticas ou participando da constituiÃÃo molecular dos fotossistemas. ProteÃnas com papel protetor contra o estresse, como chaperonas e enzimas do estresse oxidativo (SOD) foram identificadas, assim como a calreticulina, que provavelmente està envolvida em processos de transduÃÃo de sinal. Em adiÃÃo, foi possÃvel observar que durante a recuperaÃÃo das plantas os mecanismos de homeostase Ãs novas condiÃÃes restabeleceram os nÃveis de algumas proteÃnas, o que sugere a participaÃÃo delas no processo de aclimataÃÃo ao estresse salino. No geral, os resultados fornecem informaÃÃes adicionais que podem levar a uma melhor compreensÃo das bases moleculares da tolerÃncia ou sensibilidade de plantas de feijÃo de corda à salinidade. / In the present work, an integrated physiological, biochemical and proteomic analysis on cowpea (an economically important species) was carried out in order to understand the responses of plants to salinity. Two experiments were conducted in a greenhouse under hydroponic conditions, using two cultivars of cowpea (Vigna unguiculata) with differential tolerance to salt stress: Pitiuba (tolerant) and TVu 2331 (sensitive). In the first experiment, we evaluated the physiological (growth, gas exchange, relative water content, chlorophyll content, chlorophyll fluorescence) and biochemical (ions and organic solutes accumulations) changes induced by increasing levels of salinity (50, 75 and 100 mM NaCl), as well as the influences of stress two-dimensional (2D) protein patterns in leaves tissues. The results showed the existence of contrasting responses between the cultivars studied, especially with respect to inhibition of shoot growth and the accumulation of compatible solutes, which were higher in TVu. However, salinity did not alter the fluorescence parameters in both genotypes. The global analysis of 2D protein patterns showed that increasing levels of NaCl differentially alter the expression pattern of proteins in leaves, with larger changes in dose 100 mM NaCl. Nevertheless, in the second experiment, the moderate dose of salt with 75 mM NaCl was chosen as the reference dose for the study of changes in the proteome during salt stress and recovery. Salinity altered the expression of 22 protein spots in Pitiuba. Of these spots, the identities of 10 (6 up-regulated and 4 down-regulated) were determined by liquid chromatography electro-spray ionization tandem mass spectrometry (LC-ESI-MS/MS). In TVU changes were observed in 27 spots being the identity determined for 9 (5 up-regulated and 4 down-regulated). Besides these, 5 spots that kept their rates expressions in salinity conditions in PitiÃba were also identified. The majority of the identified proteins is related to the process of photosynthesis, performing enzymatic functions or participating in the molecular constitution of photosystems. Proteins with protective role against stress such as chaperones and enzymes of oxidative stress (SOD) were identified, as well as calreticulin, which are probably involved in signal transduction network. In addition, during recovery treatments homeostasis mechanisms of plants to new conditions restored the levels of certain proteins, suggesting their participation in the process of acclimation to salt stress. Overall, results provide some additional information that can lead to a better understanding of the molecular basis of salt tolerance or sensitivity in cowpea plants.

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