*2.4. Data and Molecular Analysis*

A completely random design was used for the gamma irradiation and in vitro multiplication. All treatments were performed in triplicate. Data were subjected to an analysis of variance and Tukey test (*p* < 0.05) using IBM SPSS statistical software v21. Percentage data were transformed with the formula Y = arcsine (√(×/100)), where <sup>×</sup> is the percentage value.

A binary matrix was made with the ISSR fragment bands and recorded as present (1) or absent (0). For each primer, the percentage of polymorphism was calculated. In addition, a cluster analysis was performed using Jaccard's similarity coefficient based on similarity between sets of samples relative to matches, where 1 is similarity and 0 is divergence, and the neighbor joining (NJ) agglomeration model based on minimum evolution [33]. The donor plant was set as an outgroup. Neighbor joining clustering was performed after 1000 Bootstrap replicates. The resulting cluster was expressed as a dendrogram using the PAST software v3.04.
