## 2 Results  
### THC and CBD Quantification and Phenotyping  
The study of chemical patterns and profiling of novel cannabis germplasm grown in highly diverse climates and geographical regions are likely to valuable and informative as a starting point for future cannabis breeding programs. HPLC-derived chromatograms of the extracts from the female buds and standards solution are shown in Supplementary Figure S1. Variation of THC and CBD composition of cannabis plants derived from 20 locations indicate that this collection is diverse in THC and CBD both within and between populations, and that phenotypes characterized by THC/CBD values could be classified into three classes: Type I (THC-predominant), Type II (approximately equal proportions of THC and CBD), and Type III (CBD-predominant). Furthermore, studied populations were completely different in terms of many morphophysiological features, which is beyond the scope of this study, but the plants of three unique dwarf and early flowering populations with compact inflorescence including Pir-01, Sqz-01, and Naq-01 were all located chemically in the group of Type I. Additionally, the plants of location Ard-01, which was morphologically distinct in terms of inflorescence structure, seeding time, and seed size, surprisingly, it failed to classify into either THC-predominant or CBD-predominant groups. The majority of plants of this population with almost equal concentration of THC and CBD, hereafter, a ratio of THC/CBD\(\approx 1\), were defined as Type II. The different structure of female inflorescences and buds in studied populations is presented in Figure 1.  
Our results illustrated that maximum THC was observed in the plants collected from locations Dez-02 and Dez-01 distributed in the south-west areas. The minimum THC was quantified from plants from the location Sam-01, the maximum CBD was obtained from the plants of location Rmhz-01, and the minimum CBD was obtained from Pir-01 (Figures 2 and 3a). A significant variation in THC content from 0.02% to 15.66% and CBD from 0.03% to 15.89% (% DW) were observed in this collection (Figure 2). The distribution of THC and CBD concentration (%) for each location is shown in Figure 2. A greater THC/CBD amount can be recorded due to high THC or low CBD levels. The populations of this collection with a prevalence of THC and THC/CBD content well beyond 1.0, including Pir-01, San-01, Sqz-01, Zah-01, Qzv-01, Naq-01, Abh-01, Krmn-01, Krsh-01, Bsh-01, Bam-01, Dez-01, and Dez-02, are defined as Type I. Additionally, six populations, including Nhv-01, Ban-01, Sir-01, Rmhz-01, Ark-01, and Sam-01, with a prevalence of CBD, hereafter, THC/CBD well below 1.0, were considered CBD- predominant populations [8; 29; 30; 31; 32; 33; 40]. Among CBD-predominant populations, all plants of population Sam-01 were characterized with THC level \(\leq\)0.3%. The phenotype of each population based on THC/CBD ratio is  
Figure 1: Diversity in female infloresence of 20 natural cannabis populations distributed across Iran from west and north-west to south-west and east and south-east. given in Figure 3a. As we observed THC and CBD diversity both between and within studied populations, so did we measure the abundance (%) of each phenotype (Type I, Type II, and Type III) per population in terms of the ratio of THC/CBD (Figure 3b). In order to define the phenotype of a population, we have considered the phenotype that is dominant in that population (Figure 3a); then, we have shown the abundance of all existed phenotypes in that population as well (Figure 3b). All plants of population Ard-01 had approximately equal proportions of THC and CBD, and three populations of Ban-01, Ark-01 and Sam-01 were 100% CBD- predominant and all their individuals assigned to the group of Type III (Figure 3b).