IN VITRO BULBLET REGENERATION FROM IMMATURE EMBRYOS OF ENDANGERED AND ENDEMIC MUSCARI AZUREUM

A high frequency of bulblet regeneration was achieved for the endemic and endangered ornamental plant Muscari azureum using immature embryos. Immature embryos of M. azureum were cultured on a callus induction medium consisting of N(6) mineral salts and vitamins, 400 gL(-1) casein + 40 gL(-1) sucrose + 2 mgL(-1) L-proline, 2 mgL(-1) 2,4-D and 2 gL(-1) Gelrite. Then the embryogenic callus clusters were transferred to a bulblet induction medium consisting of MS mineral salts and vitamins containing different concentrations and combinations of BAP, KIN, TDZ, Zeatin, IAA, NAA, 30 gL(-1) sucrose and 7 gL(-1) agar. Prolific bulblet multiplication (over 13 bulblets/embryo) was achieved from immature embryos after 5-6 months of culture initiation. Well-developed bulblets were excised and individually rooted on 1/2 strength MS medium supplemented with 1 mgL(-1) IBA, 0.5 gL(-1) activated charcoal, 20 gL(-1) sucrose and 6 gL(-1) agar and acclimatized.