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In an effort to optimize tissue culture responses of Italian ryegrass(Lolium multiflorum Lam.) for future genetic manipulations to improve forage characteristics, the effects of culture medium supplements on tissue culture responses were investigated with mature seeds of three cultivars, ´Jeanne´, ´Florida-80´ and ´Metro´, as explant tissues. For all explants, MS medium containing 5㎎/L 2,4-D was optimal for embryogenic callus induction from mature seed and had a strong effect on successive plant regeneration. The optimal concentration of dicamba for the induction of embryogenic callus from mature seeds was 7㎎/L. The highest plant regeneration frequency was observed when embryogenic callus was transferred to N6 medium supplemented with 1㎎/L 2,4-D and 5㎎/L BA. Plant regeneration frequency of callus cultured in the dark was higher than that of cultured in the light. Casein Hydrolysate and L-proline improved both in empryogenic callus induction from mature seeds and plant regeneration. High-frequency regeneration system established in this study will be useful for molecular breeding of Italian ryegrass through genetic transformation.
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