Alfalfa Seedling Root Morphology under Alkaline Stress

A total of 5 plants of alfalfa seedlings for each replication was randomly selected from the 25 seedlings with 5 replications in each treatment after 48 h of alkaline stress. The roots of each plant seedling were scanned using an Epson Expression 10000XL (Epson America Inc., Long Beach, CA, United States). The resulting images were digitized using the WinRHIZO program according to the instructions of the manufacturer (Regent Instruments Canada Inc., Ville de Québec, QC, Canada), and the root parameters, total root length, total root surface area, total root volume, average root diameter, and root number of forks were determined. After scanning, the seedlings were cut and divided into aboveground parts and roots. The fresh weight (FW) of alfalfa seedlings was estimated after washing with deionized water and blotting on paper towel. These samples were then dried in a forced-air-driven oven at 105°C for 2 h, followed by 70°C until maintaining a stable mass prior to the determination of dry mass (Zhang et al., 2017 (link)).