Nothofagus is the main component of southern South American temperate forests. Overexploitation in the past has led to the loss of 40% of the original distribution range. Genetic diversity as well as biological processes shaping the distribution of the genetic variation (e.g. gene flow) constitutes basic knowledge for the implementation of conservation measures and for the definition of Evolutionary Significant Units. Nuclear microsatellites are the marker of choice for gene flow and fine-scale genetic structure studies. We enlarged a previous set of microsatellites (SSRs) for South American Nothofagus species, with special concern to Nothofagus nervosa (Phil.) Dim. et Mil. Five new SSRs are presented with allele numbers up to 12 in a single population. The primers transferred well to five related species (N. obliqua (Mirb.) Oerst, N. glauca (Phil.) Krasser, N. dombeyi (Mirb.) Oerst, N. pumilio (Poepp et Endl.) Krasser and N. antarctica (G. Forster) Oerst, with allele numbers up to 11. The high level of polymorphism promises a sufficient power for gene flow and parentage analyses.