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Previous issue date: 2013-12-18 / Coffee is in international and Brazilian economical scenery as the second most important natural commodity. Despite that, coffee crops suffer great damage due to the coleopteran coffee borer (Hypothenemus hampei) attack leading to around US$ 500 million loss. In order to control the coffee borer, one strategy is the inhibition of the insect digestive enzyme ??-amylase by Phaseolus vulgaris amylase inhibitor 1 (??AI1). In this context, the research group of Plant-pest Molecular Interaction Laboratory developed GM coffee (Coffea arabica) plants expressing the P. vulgaris ??AI1 gene. This is a promising event to be used in coffee borer control by inhibiting starch insect digestion. However, releasing a GM crop for commercialization and human consumption is possible only after assessing food and environmental safety status, based on international and national procedures. This work is divided in two chapters. In the first chapter is presented the isolation of an amylase cDNA from H. hampei, designated AmyHha. The highest transcript levels of this amylase coincide with the feeding stages of insect, the second instar and adult stage. The Southern blot analysis showed the presence of only one copy of the AmyHha in the H. hampei genome. Finally, was carried out the construction of a structural model based on the AmyHha predicted protein sequence. In the second chapter, food safety of GM coffee crops expressing P. vulgaris ???-AI1 was evaluated. First, the evaluation of allergenic potential of GM coffee grains expressing ???-AI1 and the study of hemolytic effect and thermostability using coffee grains suggest absence of feeding risks concerning cytotoxic effects and inhibition of ???-AI1 after heat treatment. Otherwise, in vitro resistance to digestibility and in silico analysis suggest an allergenic risk of GM coffee grains due to ???-AI1 expression. Thus more investigation must be carried out in order to clarify allergenicity risks and in vivo consequences of this GM coffee grain consumption. / O caf?? destaca-se no cen??rio econ??mico nacional e internacional como a segunda maior ???commodity??? natural. No entanto, a cultura cafeeira apresenta grandes perdas monet??rias em torno de US$ 500 milh??es por ano devido ao ataque da broca-do-caf?? (Hypothenemus hampei). Uma estrat??gia para o controle do H. hampei ?? a inibi????o da enzima digestiva ???-amilase deste inseto pelo inibidor de ???-amilase 1 (???-AI1) de Phaseolus vulgaris. Assim, o grupo de pesquisa do Laborat??rio de Intera????o Molecular Planta-Praga desenvolveu plantas de caf?? Coffea arabica geneticamente modificadas (GM) expressando o gene para o ???-AI1 de P. vulgaris. Entretanto, a libera????o de plantas GM para comercializa????o e consumo humano s?? ?? poss??vel ap??s avalia????es de seguran??a alimentar e ambiental. Esta tese ?? dividida em dois cap??tulos. No primeiro cap??tulo ?? apresentado o isolamento de um cDNA de uma amilase de H. hampei ??? AmyHha. Os maiores n??veis de transcritos desta amilase coincide com as fases de alimenta????o do inseto, o segundo instar larval e a fase adulta. A an??lise por Southern blot demonstrou a presen??a de apenas uma c??pia de AmyHha no genoma do inseto, por fim realizou-se uma constru????o do modelo estrutural com base na sequencia proteica predita de AmyHha. No segundo cap??tulo que trata da avalia????o de seguran??a alimentar de plantas C. arabica GM expressando o inibidor de ???-AI1 de P. vulgaris, o estudo de efeito hemol??tico e termoestabilidade com os gr??os de caf?? GM sugerem a aus??ncia de risco alimentar quanto aos efeitos citot??xicos e inibi????o pelo ???-AI1 ap??s tratamento t??rmico. J?? a resist??ncia ?? digestibilidade in vitro e as an??lises in silico sugerem um potencial risco alerg??nico dos gr??os de caf?? GM devido ?? express??o do ???-AI1. Entretanto, mais investiga????es devem ser realizadas para maiores esclarecimentos quanto ao risco de alergenicidade dos gr??os de caf?? GM expressando ???-AI1 e as consequ??ncias de seu consumo in vivo.
| Identifer | oai:union.ndltd.org:IBICT/oai:bdtd.ucb.br:tede/2084 |
| Date | 18 December 2013 |
| Creators | Bezerra, Caroline de Andrade |
| Contributors | S??, Maria F??tima Grossi de |
| Publisher | Universidade Cat??lica de Bras??lia, Programa Strictu Sensu em Ci??ncias Gen??micas e Biotecnologia, UCB, Brasil, Escola de Sa??de e Medicina |
| Source Sets | IBICT Brazilian ETDs |
| Language | Portuguese |
| Detected Language | English |
| Type | info:eu-repo/semantics/publishedVersion, info:eu-repo/semantics/doctoralThesis |
| Format | application/pdf |
| Source | reponame:Biblioteca Digital de Teses e Dissertações da UCB, instname:Universidade Católica de Brasília, instacron:UCB |
| Rights | info:eu-repo/semantics/openAccess |
| Relation | -2827197273900952156, 500, 500, 600, -6392058866414562720, -5518144268585252051 |
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