杜仲胚轴、子叶直接诱导不定芽及再生体系的建立

Direct inducement of adventitious bud by hypocotyl and cotyledon and establisment of regeneration system of Eucommia ulmoides

目的探索杜仲胚轴、子叶不定芽诱导条件,建立杜仲再生体系。方法分别以杜仲种胚、胚轴切段、子叶切段、芽为外植体,将它们接种到一系列含有不同激素质量浓度的培养基中。结果在外植体的选择上,胚轴优于子叶。对于杜仲胚轴的伸长生长,以MS培养基附加2.0mg/LGA效果最好。在含有0.1mg/LNAA及0.8mg/L6-BA的MS培养基中,杜仲胚轴可直接诱导产生不定芽,诱导率达到47%。在壮芽及生根培养中,分别采用附加1.0mg/L6-BA、0.1mg/LNAA的MS培养基及附加1.5mg/LIBA、20mg/L蔗糖的1/2MS培养基效果最好,经28、14d,芽苗生长系数及生根率分别达到2.48和91.7%。结论建立了稳定高效的杜仲再生体系,为外源基因导入杜仲奠定了基础。

Objective To explore the buds inducement condition of hypocotyls and cotyledons and establish the regeneration system of Eucommia ulmoides. Methods Taking embryos and hypocotyl slices, cotyledon pieces and shoots from E. ulmoides as explants, to inoculate them in a series of media with hormone at various concentrations. Results In selection of explants, hypoeotyls were better than cotyledons. The MS medium containing 2.0 mg/L GA was the most favourable for hypocotyl elongation. Being cultured in MS medium containing 0. 1 mg/L NAA and 0. 8 mg/L 6-BA, hypocotyl slices could be induced to produce adventitious buds directly and the inducement ratio was up to 47%. The optimum media for shoot strengthening and root inducing were MS＋1.0 mg/L 6-BA and 0. 1 mg/L NAA, and 1/2 MS＋1.5 mg/L IBA and 20 mg/L sucrose. Bud growth coefficient and root inducting ratio reached 2. 48 and 91.7% in 28 d and 14 d, respectively. Conclusion A stable regeneration system for E. ulmoides with higher efficient has been established, so as to lay a way for transferring the exogenous gene into E. ulrnoides.