*4.1. Plant Materials*

The study material comprised 12 tetraploid CWG lines consisting of six breeding lines, three cultivars, and three genebank accessions (Table 1). These accessions were acquired from USDA-ARS plant germplasm system, Plant Gene Resources of Canada (PGRC), and the joint forage breeding program of the University of Saskatchewan and Agriculture and Agri-Food Canada (AAFC). For ease of interpretation, all the acquired material will be referred to as lines, rather than accessions, in this study. Seeds of each line were grown for six weeks in the greenhouse at the Saskatoon Research and Development Centre, AAFC, under the following growth conditions: 16 h photoperiod at 22 ◦C and 8 h dark at 16 ◦C. Young leaf tissues were collected from 16 randomly selected plants for each of the lines and stored at −80 ◦C prior to DNA extraction. A total of 192 genotypes from the 12 tetraploid lines, listed in Table 1, were used for bioinformatics and genetic diversity analyses.
