High frequency plant regeneration from mature seedderived callus of Italian ryegrass (<i>Lolium multiflorum</i>) cultivars

Abstract

In the present study, we have developed a high-frequency plant regeneration system for Italian ryegrass via callus culture using mature seeds as explants. Optimal embryogenic callus induction was found to occur in MS medium containing 5 mg l-1 2,4-D, 0.5 mg l-1 BA, 500 mg l-1 L-proline, 1 g l-1 casein hydrolysate, 30 g l-1 sucrose, 7 mg l-1 AgNO3, 2 mg l-1 CuSO4 and solidified with 3 g l-1 Gelrite. The highest regeneration rate was obtained in MS medium containing 1 mg l-1 2,4-D, 5 mg l-1 BA, 500 mg l-1 L-proline, 1 g l-1 casein hydrolysate, 1 mg l-1 thiamine-HCl, 30 g l-1 sucrose, 7 mg l-1 AgNO3, 2 mg l-1 CuSO4 and solidified with 3 g l-1 Gelrite. By using the most effective treatment determined for each parameter, the highest rates of embryogenic callus formation (48.9%) and regeneration (47.6%) were obtained with the Hwasan 101 cultivar. The overall plant regeneration rates of the examined cultivars ranged from 7.5 to 23.2%. Thus, optimization of regeneration frequency using mature seeds as explant material may offer a simple and efficient protocol for Italian ryegrass that may improve molecular breeding of this species.