一种卷丹愈伤组织的诱导与增殖方法

A Method of Callus Induction and Proliferation for Lilium lancifolium Thunb.

卷丹难以诱导形成块状愈伤组织,以卷丹珠芽、鳞茎鳞片、鳞茎盘和无菌小鳞片为外植体,探讨不同激素和光照条件对愈伤组织诱导与增殖的影响。结果表明,诱导卷丹愈伤组织最佳材料为无菌小鳞片;最佳培养基配方为MS+1.00 mg/L 6-BA+0.15 mg/L NAA+0.20 mg/L KT+0.05 mg/L 2,4-D+30 g/L蔗糖,最适光照条件是700~900 lx。当培养条件为最优时,20 d左右卷丹愈伤诱导率达到71%,愈伤直径大于1 cm。在黑暗条件下,愈伤组织继代增殖最佳培养基为MS+1.00 mg/L 6-BA+0.15 mg/L NAA+0.20 mg/L KT+0.10 mg/L2,4-D+30 g/L蔗糖,30 d后增殖率达到80.83%,增殖倍数为4.63倍。本研究提供了一种快速获得卷丹大量愈伤材料的方法,为卷丹的离体快繁和遗传育种改良提供技术支持。

To induce a large number of calli for Lilium lancifolium Thunb.is hard to perform.Here the effects of different hormones and light conditions on callus induction and proliferation were studied by using the bulbil,bulbous scale,bulbous disc and small sterile bulblet scale as explants.The results showed that the best materials for callus induction were small sterile bulblet scale,the best medium was MS+1.00 mg/L 6-BA+0.15 mg/L NAA+0.20 mg/L KT+0.05 mg/L 2,4-D+30 g/L sucrose,and the best condition of light was 700~900 lx.The callus induction rate can reach 71%and the callus size was more than 1 cm in 20 days under the best cultural conditions.The best medium for callus proliferation was MS+1.00 mg/L 6-BA+0.15 mg/L NAA+0.20 mg/L KT+0.10 mg/L2,4-D+30 g/L sucrose.The best growth coefficient reached 80.83%,and proliferation rate was 4.63 times in 30days respectively.The research provides a rapid method to obtain a large number of calli and lays the foundation for in vitro rapid propagation and genetic breeding improvement of L.lancifolium.