Identificação molecular de cultivares de soja (Glycine max L. Merrill) utilizando um sistema semi-automatizado de genotipagem

• Autor(a) principal: Passianotto, André Luiz de Lima
• Data de Publicação: 2010
• Tipo de documento: Dissertação
• Idioma: por
• Título da fonte: Repositório Institucional da Universidade Estadual de Maringá (RI-UEM)
• dARK ID: ark:/35916/0013000002x0t
• Texto Completo: http://repositorio.uem.br:8080/jspui/handle/1/1398
• topic: Glycine max; Microssatélites; Validação; Marcadores moleculares; Eletroferograma; Brasil.; Glycine max; Microsatellite; Validation; Brazil.; Ciências Agrárias; Agronomia

The cultivar protection is realized based on morphological, physiological and biochemical descriptions, characteristics sensitive to environmental variations. When self pollinating species with low genetic variability are considered, as soybean, the distinction between cultivars using the standard traits demands lots of effort and it?s not efficient. In this sense, one strategy to improve the characterization is to analyze directly the DNA. Molecular markers are ideal tools t perform this work, once they are not environmental sensitive and can be applied in any stage of plant development. Among molecular markers, microsatellites are distributed through all plant genome, specific, multiallelics and co-dominant. Therefore, by the above, this work aimed to develop a semi-automated system, by employing fluorescent SSR markers, that would allow genetically distinguish soybean cultivars. For this, forty ? eight soybean cultivars were selected in the germoplasm bank from Embrapa ? Soybean. Seed were germinated in pots and kept in green house until the first leaflet appearance (V3 stage) which was collected from each cultivar in a bulk methodology and storage in -80°C. Posteriorly, DNA extraction was done and analyzed using 23 pairs of microsatellites primers. Sense strains were labeled with blue (6FAM), green (HEX) or yellow (NED) fluorescence. The analyses from results fragments was carried out in a automatic sequencer ABI PRISM Genetic Analyser® 3100 (Applied Biosystem, Foster City, CA) and the software GeneMapper® (AB-PEC,Foster City, CA) was applied to visualize the alleles exactly and to emit data automatically. The electroferograms obtained allowed the identification informative loci and its alleles.In total of 201 alleles were observed, with an average of 8.73 alleles per loci. The most informative loci were Sat_038 and Satt009, with 12 alleles each and the less informative were Satt042, Satt045, Satt002 and Satt233 with six alleles each. The loci Sat_038, Satt612, Satt181, Satt540, Satt009 and Satt005 allowed the identification and differentiation of all group of the 48 soybean cultivars studied, being named loci differentiator. Based in these loci it was possible perform the identification of genotypes and consequently elaborate genetic tags. Such a system will be used in the characterization of new cultivars, assisting the protection procedure, as well as in cases to analyzed seed genetic purity when visual analyses were doubtful.