GBS of Medicago sativa
Breeding populations containing 179 breeding lines developed from 3 families by Forage Genetics International (FGI) for VW resistance. The populations were propagated by stem cuttings and three clones from each individual were evaluated for resistance to Verticillium wilt in a greenhouse assay as described below. The cloned plants were removed from propagation flats, and transplanted into a pot containing commercial potting soil (Pro-Mix). The stems were clipped to a length of approximately 5 cm once plants began to flower to generate clones of approximately the same size. The cloned populations were placed in a randomized complete block design with three replications and with each replication consisting of one plant at least. Cloned plants from the population were inoculated with conidia from V. alfalfae isolates (N-29, Nyvall and 34, provided by the S&W Seed Co.) mixed with equal amount and inoculated to the plants according to the standard protocol (Grau et al. 1991). After incubation, Plants were evaluated for disease reaction of foliar symptoms based on the standard protocol (Grau et al. 1991). The disease resistance scores of three replications from the same individual were statistically analyzed to obtain least square means for further analysis.