欧洲百合愈伤组织诱导及植株再生体系的建立

以欧洲百合(Lilium martagon)无菌苗鳞片为外植体, 探讨不同植物激素组合及光暗培养条件对愈伤组织诱导、增殖和再生不定芽的影响, 进而建立欧洲百合高效再生体系。结果显示, 诱导愈伤组织的最佳培养基为MS+0.2 mg?L^-1 TDZ+0.5 mg?L^-1 NAA, 诱导率为77.14%。在添加TDZ和NAA组合的培养基中进行继代培养, 愈伤组织极易褐化, 胚性活性下降; 采用添加6-BA和NAA组合的培养基可改善愈伤组织的褐化现象, MS+0.5 mg?L^-1 6-BA+0.1 mg?L^-1 NAA是愈伤组织增殖的最佳培养基, 增殖指数为2.93, 表明6-BA在愈伤组织状态维持中起关键作用。暗培养条件下愈伤组织的诱导率、增殖指数和芽再生系数最高, 分别可达77.14%、2.93和5.43, 且愈伤组织生长状态较好, 不定芽生根正常。研究建立的欧洲百合高效再生体系对于百合种质资源保存、基因工程育种及在国内的推广应用具有重要意义。

Callus Induction and Establishment of a Plant Regeneration System with Lilium martagon

We studied the effectiveness of different combinations of plant growth regulators and the light/dark condition on callus induction, proliferation and differentiation of bulb scales in Lilium martagon and established a high-frequency regeneration system. The optimal medium for callus induction was MS+0.2 mg?L^-1 TDZ+0.5 mg?L^-1 NAA, with embryogenic callus induced from 77.14% bulb scales. For callus proliferation, MS medium containing TDZ and NAA produced a browning appearance with the callus lost embryogenic ability quickly; therefore, 6-BA was applied instead of TDZ. Generation of buds from callus was reduced with 6-BA concentration increasing, which indicates that 6-BA plays a key role in keeping meristematic cells of callus from differentiating. The optimal medium for callus proliferation was MS+0.5 mg?L^-1 6-BA+0.1 mg?L^-1 NAA, with the proliferation coefficient 2.93. Dark treatment led to the highest callus induction rate, proliferation coefficient and number of buds/callus (77.14%, 2.93 and 5.43, respectively); callus showed good growth and the buds could grow roots and develop complete plants, which suggests that the dark condition is more appropriate for in vitro regeneration than the light condition. The rapid propagation system of L. martagon established in this study offers insights into germplasm protection, genetic transformation and application of L. martagon in China.