Establishment regeneration system of Russian variegated alfalfa

To realize the use of excellent germplasm resource and lay the foundation of alfalfa genetic improvement. This study took MS solid medium as basic medium and cotyledons and hypocotyls of Medicago sativa subsp. varia as explants to discuss the effects of different phytohormone types and combinations on callus induction and differentiation. The results showed that hypocotyls was more suitable to induce callus than cotyledons as the explant of Russian variegated alfalfa and the best induced medium with hypocotyls as the explant was MS+1.0 mg·L-1 2,4-D +0.5mg·L-1 6-BA+0.5 mg·L-1 KT; The best induced medium with cotyledon as the explant was MS+1.0 mg·L-1 2,4-D+1.0 mg·L-1 6-BA+1.0 mg·L-1 NAA. The best differentiation medium was 0.5 mg·L-1 KT +0.3 mg·L-1 NAA, and differentiantion ratios of hypocotyls and cotyledons were 83.3% and 91.1%, respectively. The optimal seedling formation medium was 1/2MS+1.0 mg·L-1 IBA, and the ratio was 96.7%.