DESENVOLVIMENTO E CARACTERIZAÇÃO DE MARCADORES MICROSSATÉLITES PARA REGIÕES ALVOS DE RESISTÊNCIA À Meloidogyne ssp. em Phaseolus vulgaris

Abstract

The use of molecular markers has grown exponentially in recent years, especially in studies of plant populations genetics, when it comes to the conservation of important species or that are inserted in biomes that need to be preserved. As an example, we have the bean (Phaseolus vulgaris) cultivated in the Cerrado biome, being exposed to different types of soils, climates and pathogens. Among the root pathogens, the gill nematodes stand out, being considered pest of the common bean. Therefore, the target was develop and characterize microsatellite markers that confer resistance to the nematode Meloidogyne spp. in common beans. For this, QTL sequences were selected in the NCBI database because they presented genes homologous to those of soybeans because they had resistance to the pathogen in question and were identified for the bean using the BLAST program at Phytozome. Nine primers were designed using the Primer3 software. The DNA extraction of common bean seeds was done using a 10% SDS extraction protocol. Quantification was performed on 0.8% agarose gel electrophoresis, stained with ethidium bromide (0.5%/ml) in 1x buffer solution (TBE). For amplification and PCR optimization, the DNA of a single genotype was used, using thermocycler. Functional PCRs were submitted to vertical electrophoresis in polyacrylamide gels (4%) and stained with silver nitrate. Transferability allows to increase the efficiency in the transfer of genomic information through EST homology of genes of interest. Nine of 26 EST-SSR bean candidates distributed in seven different genes conferring nematode resistance to gall in soya are available for validation.