VALIDACIÓN Y ADAPTACIÓN DE LA TÉCNICA DE MICROSATÉLITES PARA EL ANÁLISIS GENÉTICO DE Vochysia guatemalensis Donn. Sm.

Recent advances in clonal forestry in Costa Rica and internationally as well, generates new needs. It is now mandatory to be able to guaranty genetic purity in clonal collections; to control the inbreeding and genetic variability in breeding populations, among other applications. Gene markers usage and late developments, like with microsatellites (SSR), constitute a new and invaluable tool for guaranteeing and certifying genetic identity in gene conservation and breeding programs in forestry. In this work is presented the adaptation and validation of microsatellites in Vochysia guatemalensis, that had been previously developed for V. ferruginea. DNA extraction method, based on CTAB, allowed a high quality material, determined through agarosa gels (1% concentration). Initial amplification evaluation, based on a previous protocol for Vochysia ferruginea, showed non expected results, therefore, a new protocol for V. guatemalensis was needed and developed. The validation process resulted in the elimination of one of the five primers analyzed (A1-26). Alleles size found was different from reported in V. ferruginea. The closer to V. ferruginea average allele size (in bp) was determined for primer A1-35. The four investigated loci showed high degree of polymorphism (4, 4, 3 and 2 alleles respectively), which allowed them to be utilized as appropriated primers for V. guatemalensis.