Winrhizo pro 2009c

At 24 DAT, all plants were harvested by separating the stem of each plant at ground level from its root system. Roots were then removed from the pots, placed on a wire screen, and washed thoroughly to remove the soil medium, using a moderate hydro flow speed and exercising maximum caution to avoid damage to the root structures. The longest root length was recorded using a meter ruler for each plant root. The individually cleaned root system was scanned using a Epson Expression 11000XL scanner, attached to a computer system. The individually cleaned root structures were placed onto a waterproof Plexiglas tray (40-cm length × 30-cm width) filled with approximately 5 mm of water and fitted onto the scanner. The roots were submerged, and the crossings and tips were spread using a small paintbrush to avoid overlapping. The acquired gray-scale root images were obtained through a high accuracy setting (resolution of 800 by 800 dpi) for the parameters measured by the WinRHIZO Pro 2009C software (Regent Instruments, Québec, Canada). The software calculated the following components: total root length, root surface area, average root diameter, root volume, number of root tips, forks, root crossings, root length density (ratio of total root length to root volume), and root to shoot percentage (ratio of root weight to shoot weight).

After the above-ground plant parts were removed, root systems and soil media were gently removed from the PVC pots and washed with a gentle stream of water over a wire mesh sieve to remove soil media until the roots were clean. Individual root systems were floated in a 400 × 300 cm acrylic tray filled with 5 mm water. Once placed on the trays, the roots were carefully untangled using plastic forceps to minimize roots’ overlap to ensure quality imagery. An Epson Expression 11000XL scanner captured root morphology images at a 800 dpi resolution (Epson America, Inc., Long Beach, CA, USA). These images were analyzed by WinRHIZO Pro 2009C software (Regent Instruments, Inc., Québec, QC, Canada). The digitized output from the analysis quantified the multiple root growth and development parameters for each plant: root tips (RT, no plant⁻¹), root forks (RF, no plant⁻¹), total root length (TRL, cm plant⁻¹), root surface area (RSA, cm² plant⁻¹), and root volume (RV, cm³ plant ⁻¹)