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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.
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Seed ontogeny and proteomic analysis of the internal integument of Jatropha curcas L. (Euphorbiaceae) / Ontogenia da semente e anÃlise proteÃmica do integumento interno de Jatropha curcas L. (Euphorbiaceae)

Emanoella Lima Soares 23 February 2015 (has links)
Conselho Nacional de Desenvolvimento CientÃfico e TecnolÃgico / The economic importance of J. curcas is due to the high amount of oil accumulated in their seeds, specifically in the endosperm. In this context, the events related to the endosperm formation and the accumulation of toxic compounds in the seeds could help in the production of varieties that produce more oil and are less toxic. Thus, anatomical, proteomic and gene expression analysis were made with J. curcas seeds to contribute with the knowledge about physiologic events related with the formation of endosperm and the processes of synthesis and accumulation of phorbol esters. The anatomical analysis showed the maternal tissue responsible for the nutrients transfer to the endosperm and contributed to fill the gaps present in the literature about the anatomy of J. curcas seeds. The inner integument is the maternal tissue that presents the vasculature of the mother plant and is the responsible for the nutrition of the endosperm through the process of programmed cell death (PCD). Based on this result proteomic analysis of two anatomically different regions of the inner integument, named as proximal and distal, were performed. The identified proteins indicated that the inner integument pass through PCD in different degrees in the proximal and distal regions and that in this tissue there is a spatial distribution between catabolism and anabolism. In the proximal region especially hydrolases were identified and in the distal region storage proteins and proteins of biosynthetic pathways were prevalent. The gene expression analysis had two purposes: first, verify wether the transcripts levels of the genes likely involved in PCD, whose proteins were identified and quantified in the proteomic analysis, were related to the protein abundance and second, identify the site of synthesis of the committed enzyme in the phorbol esters biosynthesis, the casbene synthase, through a study with root, leaf, embryo, inner integument and endosperm and 14 putative genes of the mentioned enzyme. The results indicated that the proteins related to PCD must be controlled at transcript level. Nine putative genes from casbene synthase were expressed in roots and one of them was expressed in the leaf and embryo. This work denotes the role of PCD in the inner integument to the development of the seed and also demonstrates the transitory nature of storage of this tissue. In addition gives notation that the seed, despite of accumulates phorbol esters, do not synthesizes these compounds, but probably import them from roots. / A importÃncia econÃmica da espÃcie J. curcas deve-se a grande quantidade de Ãleo acumulada em suas sementes, especificamente no endosperma. Os eventos relacionados à formaÃÃo do endosperma e ao acÃmulo de compostos tÃxicos nas sementes podem auxiliar na produÃÃo de variedades com maior produÃÃo de Ãleo e menos tÃxicas. Visando isso, foram realizadas anÃlises anatÃmicas, proteÃmicas e de expressÃo de genes nas sementes de J. curcas para contribuir com o entendimento sobre os eventos fisiolÃgicos associados à formaÃÃo do endosperma e aos processos de sÃntese e acÃmulo dos Ãsteres de forbol. A anÃlise anatÃmica mostrou o tecido maternal responsÃvel pela transferÃncia de nutrientes para o endosperma e contribuiu para resolver lacunas existentes na literatura sobre a anatomia de sementes de J. curcas. O integumento interno à o tecido maternal que apresenta a vascularizaÃÃo da planta-mÃe e à o responsÃvel pela nutriÃÃo do endosperma atravÃs do processo de morte celular programada (PCD). Com base nesse resultado, realizou-se uma anÃlise proteÃmica do integumento interno na regiÃo proximal e distal ao endosperma, uma vez que as caracterÃsticas celulares indicativas de PCD nesse tecido se apresentavam diferentes nessas regiÃes. As proteÃnas identificadas indicaram que o integumento interno sofre PCD em diferentes graus nas diferentes regiÃes e que nesse tecido hà uma distribuiÃÃo espacial entre catabolismo e anabolismo, pois na regiÃo proximal identificaram-se principalmente hidrolases e na regiÃo distal proteÃnas de reserva e de vias biossintÃticas. A anÃlise de expressÃo de genes teve dois propÃsitos: primeiro, verificar se os nÃveis de transcritos de genes possivelmente relacionados à PCD, cujas proteÃnas foram identificadas e quantificadas na anÃlise proteÃmica, estavam relacionados à abundÃncia de proteÃnas e o segundo foi identificar o local de sÃntese da enzima comprometida na biossÃntese dos Ãsteres de forbol, a sintase do casbeno, atravÃs de um estudo com raiz, folha, embriÃo, integumento interno e endosperma e 14 genes candidatos à enzima mencionada. Os resultados indicaram que as proteÃnas relacionadas à PCD devem ser controladas no Ãmbito transcricional. Nove genes candidatos à sintase do casbeno foram expressos em raiz e um deles foi expresso na folha e no embriÃo. Este trabalho evidencia o papel da PCD no integumento interno para o desenvolvimento da semente, alÃm de demonstrar a natureza transitÃria de reserva desse tecido. Adicionalmente, dà indÃcios de que a semente, apesar de acumular Ãsteres de forbol, nÃo sintetiza esses compostos, mas possivelmente os importa das raÃzes.

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