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Three natural populations of Copaifera langsdorffii Desf. were studied by isoenzyme electrophoresis to determine the genetic structure, levels of genetic variability, gene flow and effective size of the populations. The populations are located in the countyof Lavras, in Southern Minas Gerais state and are represented by three types of vegetation (Savanna, Riparian forest and Semi-deciduous forest). Samples of leaf tissue were taken from 20 adult plants from each population and from 400 progenies for isoenzyme electrophoresis analysis. Five enzymatic systems were evaluated in adults (24 alleles average) and progenies (29 alleles total). The proportion of polymorphic loci (0,95) among populations was 72.73% to 87.50%. The average number of alleles per locus was 2.2 to 2.5 and the expected heterozygosity (He) was 0.368 to 0.435. The estimated parameters revealed inbreeding in the adult populations (f= 0.130) and excess of heterozygotes in the progenies (f=-0.033). The genetic diversity index of the populations was 0.142 for the Savanna-Riparian forest, 0.162 for the Riparian Semi-deciduous forest, and 0.073 for the Savanna-Semi-deciduous forest. The gene flow, estimated by the number of migrants (Nm), was 0.79 for Savanna-Semi-deciduous forest, 0.38 for Savanna-Riparian forest, 0.32 for Riparian-Semi-deciduous forest and 0.41 for all analyzed populations. The population from the Riparian forest has a relatively high genetic variability and is valuable for genetic conservation of C. langsdorffii.