The choice of the best ISSR primers is an essential step in obtaining robust data for genetic studies and for this selection, the use of statistical indices should be prioritized by visual evaluation. In this study, the evaluation of PIC, MI, and RP data allowed the selection of the 10 most informative primers for genetic variability studies in A. flaccida (Table 3). These 10 primers amplified 190 polymorphic loci in the seven A. flaccida populations. This number is considered adequate because according to Colombo et al. , seven to 30 primers that amplify between 50 and 200 polymorphic fragments are sufficient for genetic diversity studies within a species.