Caracteriza????o gen??mica e molecular do guaranazeiro (Paullinia cupana var. Sorbilis)

• Autor(es): Freitas, Danival Vieira de
• Data: 2015
• Origem: Oasisbr
• Assunto(s): Citogen??tica; Citometria de fluxo; Genoma funcional; Express??o g??nica; Paullinia cupana var. Sorbilis (Guaranazeiro); CI??NCIAS BIOL??GICAS
• Tipo de Documento: Tese de doutoramento
• Idioma: Português

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N??o informada

Paullinia cupana is originary from Amazon and is divided in two different varieties, P. cupana var. cupana (Venezuela and Colombia) and P. cupana var. sorbilis (Brazil), this last one is the true guarana plant. The few studies on this plant are concentrated, mainly, in its bothanical aspects and effects of its seed extract over some medical experimentation. The main objective of this work was to amplify knowledgment over those informations about the guarana plant, moreover by the means of genetic molecular. Aiming the following propositions: (1) to determine the guarana plant (P. cupana var sorbilis) caryotype and its genome size; (2) to characterize and validate sequences expressing in the guaran?? berries, gathering especial information on those related with caffeine synthesis (N-methil-transferases); (3) to determine its (Nmethil- transferases) copy number in the plant genome; (4) to determine its (N-methiltransferases) expression patterns in different organs/tissues. The results showed an elevated number of chromosomes (210) and DNA content (2C = 22.5 pg DNA/cell). The caryogram analysis presented two sets of chromosomes, one tetraploid and other hexaploid. The transcripts sequencing yield a 15,387 ESTs databank, grouped in 2,628 contigs and 5,969 singletons. Among these are enzymes genes involved in caffeine synthesis, the so called caffeine sintases. The analysis of these guarana sequences (denominated GNMT) shows it is homologous to N-methyltransferases of other species such as coffee (Cofea ar??bica and C. canephora), cocoa (Theobroma cacao), and green tea (Camellia sinensis). Regarding to the number of genome copies, the guarana plant presented three times more than its native wild plants (the guaranaranas, Paullinia sp) correlates. Fruits posses the highest level of GNMT transcript expression when it reach 70 days of development age, increasing the rate as berries ripening process occurs. These results are especially interesting and can collaborate with the experimentations in progress course which are proving the healing properties attributed to this plant and its seed powder extract.

A esp??cie Paullinia cupana origin??ria da Amaz??nia, ?? dividida em duas variedades, P. cupana var. cupana (Venezuela e Col??mbia) e P. cupana var. sorbilis (Brasil). Os poucos estudos com o guaranazeiro concentraram, principalmente, nos seus aspectos bot??nicos, no seu melhoramento e em testes experimentais sobre o efeito de seu extrato para fins terap??uticos. O objetivo principal do presente trabalho foi ampliar o conhecimento sobre as informa????es b??sicas do ponto de vista gen??tico e molecular do guaranazeiro. Deste modo foi proposto: (1) determinar o cari??tipo e o tamanho do genoma da planta (P. cupana var sorbilis); (2) caracterizar e validar seq????ncias expressas em seus frutos, e em especial as das enzimas relacionadas com a s??ntese de cafe??na (N-metiltransferases); (3) determinar o seu n??mero de c??pias; (4) determinar o padr??o de express??o em diferentes ??rg??os/tecidos. A var. sorbilis apresentaou elevado n??mero de cromossomos (2n = 210) e conte??do de DNA (2C = 22,5 pg DNA/c??lulas). A an??lise do cariograma mostrou que os cromossomos est??o agrupados em dois conjuntos, um tetrapl??ide e outro hexapl??ide. O seq??enciamento de transcritos gerou um banco com 15.387 ESTs, distribu??das em 2.628 contigs e 5.969 singletons. Entre estas est??o as enzimas que participam das vias de s??ntese da cafe??na sintase, sendo o melhor hit com as sequ??ncias de N-metiltransferases, aqui denominadas GNMT. As an??lises indicam que a sequ??ncia GNMT ?? hom??loga as N-metiltransferases da via biossint??tica da cafe??na de outras esp??cies tais como caf?? (Coffea arabica and C. canephora), cacau (Theobroma ca????o), e ch?? verde (Camellia sinensis). O n??mero de c??pias g??nicas da N-metiltransferase da GNMT presentes no genoma do guaranazeiro foi cerca de tr??s vezes mais c??pias que os esp??cimes selvagens (guaranarana, Paullinia sp). O fruto do guaranazeiro com 70 dias de desenvolvimento tem o maior n??vel de transcritos da GNMT, sendo que o n??mero de transcritos aumentou gradualmente com o processo de matura????o do fruto do guaranazeiro. Os resultados aqui discutidos s??o especialmente interessantes e podem colaborar com a experimenta????o cient??fica que est??o, aos poucos, comprovando as propriedades medicinais atribu??das ao guaran?? e seu extrato obtido do p?? de suas sementes.