Determinação da composição de bases AT e GC por citometria de fluxo em abelhas

• Ano de defesa: 2012
• Autor(a) principal: Soares, Fernanda Aparecida Ferrari
• Orientador(a): Carvalho, Carlos Roberto de
• Banca de defesa: Clarindo, Wellington Ronildo
• Tipo de documento: Dissertação
• Tipo de acesso: Acesso aberto
• Idioma: por
• Instituição de defesa: Universidade Federal de Viçosa
• Programa de Pós-Graduação: Mestrado em Genética e Melhoramento
• Departamento: Genética animal; Genética molecular e de microrganismos; Genética quantitativa; Genética vegetal; Me
• País: BR
• Palavras-chave em Português: Citometria de fluxo; Ácido desoxirribonucleico; Genoma; Abelha
• Palavras-chave em Inglês: Flow cytometry; Acid desoxirribonucleico; Genomic; Bees
• Área do conhecimento CNPq: CNPQ::CIENCIAS BIOLOGICAS::GENETICA::GENETICA ANIMAL
• Link de acesso: http://locus.ufv.br/handle/123456789/4767

The genomic size determination and AT GC base composition should be considered an important factor in the analysis to genetic material characterization, or even in studies of molecular biology and phylogenetic interpretations. Flow cytometry has emerged as a tool for quantification of DNA content and characterization of base composition in many plant and animal groups, including insects. Considering the hymenopterous, about 100 species had their DNA content determined, and 46 % of them are stingless bees. This work aimed to detail the procedure for DNA quantification and to develop methodology for determining the AT and GC base composition by flow cytometry in bees. It will increase the number of species with DNA content determined and provide an effective genomic characterization procedure. Scaptotrigona xantotricha (2C = 0.88 picograms) was used as internal standard to determinate the DNA content and base composition of Trigona hyalinata and Partamona rustica. The histograms generated in the flow cytometry showed coefficients of variation below 5.0 % and enabled the determination of the nuclear DNA content and AT GC composition. P. rustica and T. hyalinata showed 1.15 and 1.07 picograms of DNA, respectively. S. xantotricha presented 61.32 % of bases AT and 38.68 % of GC, P. rustica presented AT = 62.82 % and GC = 37.18 %, T. hyalinata presented AT = 62.40 % and GC = 37.60 %. This study details the protocol for DNA quantification by flow cytometry, contributing to increasing the number of species of DNA content determined. Also provides methodology for determining the base composition AT and GC bees using flow cytometry. This new technique can be applied to other bee species or even other hymenopteran insects, in order to provide relevant data for genomic characterization analysis.