In vitro bulblet regeneration from immature embryos of Muscari azureum

A high frequency bulblet regeneration was achieved for endemic and endangered ornamental plant Muscari azureum using immature embryos. Immature embryos of M. azureum were cultured on callus induction medium consisting of N(6) mineral salts and vitamins, 400 mg/L casein + 40 g/L sucrose + 2 g/l L-proline, 2 mg/L 2,4-D and 2 g/L gelrite. Then, embryogenic callus clusters were transferred to bulblet induction medium consisting of MS mineral salts and vitamins containing different concentrations and combinations of N(6)-benzylamino-purine (BAP), kinetin (KIN), thidiazuron (TDZ), zeatin, indole-3-acetic acid (IAA), alpha-naphthaleneacetic acid (NAA), 30 g/L sucrose and 7 g/L agar. Prolific bulblets 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 mg/l IBA, 0.5 g/l activated charcoal, 20 g/l sucrose and 6 g/l agar and acclimatized.