凤尾兰花丝诱导的组培再生体系的建立及遗传稳定性分析

Regeneration Systems for in Vitro Culture of Filament and Genetic Variation Analyses of Yucca gloriosa L.

以凤尾兰(Yucca gloriosa L.)花丝为外植体,研究不同激素种类和配比对愈伤组织诱导和分化的影响,以及活性炭对外植体褐化率的影响,以期为进一步工厂化育苗提供技术支持。结果表明:以花丝为外植体不仅可以获得较高的繁殖系数,而且可以避免灭菌剂对外植体的伤害,从而有效降低组织培养的难度和污染率;凤尾兰花丝最佳愈伤诱导培养基为:MS+6-BA 5.0mg潘·L-1+KT 1.0mg·L-1+2,4-D 0.5mg·L-1,最佳分化培养基为:MS+6-BA 6.0 mg·L-1+KT 0.5 mg·L-1+NAA 0.5 mg·L-1,最佳不定芽增殖培养基为:MS+6-BA 3.0mg·L-1+KT 0.5mg·L-1+NAA 0.05mg·L-1+椰乳100mL·L-1,增殖系数达到3.5左右;利用RAPD分子标记技术,在DNA水平上对再生株进行遗传变异鉴定,未检测出遗传变异,表明再生植株能够保持遗传上的稳定性。本研究首次建立了凤尾兰花丝高效稳定的组培再生体系。

Using the filament as explants,the effects of different combinations of hormone on callus induction and differentiation,and the effects of activated carbon concentration on callus browning rate in tissue culture process of Yucca gloriosa L.were studied.Results showed that explanted filament could not only gain a higher propagation coefficient,but also reduce tissue damage from disinfectant contaminations.The best callus induction medium was MS＋6-BA 5.0 mg·L-1＋KT 1.0 mg·L-1＋2,4-D 0.5 mg·L-1;and the best callus differentiation medium was MS＋6-BA 6.0 mg·L-1＋KT 0.5 mg·L-1 ＋NAA 0.5 mg·L-1;and the best in vitro shoot proliferation culture medium was MS＋6-BA 3.0 mg·L-1 ＋KT 0.5 mg·L-1＋NAA 0.05 mg·L-1＋coconut milk 100 mL·L-1.Regenerated plantlets were tested by randomly amplified polymorphic DNA（RAPD） with no genetic variation detected.Regenerated plantlets can maintain genetic stability.This is the first report for in vitro culture and regeneration using filaments from Y.gloriosa L.