*2.1. Plant Materials and DNA Extraction*

A total number of 147 pepper (*C. frutescens*) accessions, which had been collected from 25 countries, were used in this study. The details of the samples encompassing their English names, cultivation regions, DNA concentrations (ng/uL), and the type of selected species (wild/cultivated) are summarized in Supplementary File S1. Following the sampling processes, young leaves were immediately frozen under liquid nitrogen and transferred to −80 ◦C conditions for future DNA extraction. Then, following the process described by Murray and Thompson [34], CTAB (Cetyl Trimethyl Ammonium Bromide) methods were used for genomic DNA extraction. In addition, the quality of isolated DNA was ascertained using an NanoDropOneC Microvolume UV-Vis Spectrophotometer. Finally, the DNA concentrations were adjusted to 10 to 35 ng/μL for use in the subsequent polymerase chain reactions (PCR).
