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目的:建立灰毡毛忍冬新品种‘渝蕾一号’Lonicera macranthoides Hand.-Mazz.‘Yulei1’组培快繁体系,为实现其工厂化育苗提供理论依据。方法:以‘渝蕾一号’幼嫩茎段和叶片为外植体,以MS为基本培养基,在组织培养的不同阶段添加不同浓度的NAA、6-BA、TDZ、IBA等植物生长调节物质,对愈伤组织的诱导、增殖及分化,茎段腋芽诱导、增殖及生根炼苗进行系统研究。结果:外植体的最佳灭菌时间:茎段为2~3 min,叶片为1~2 min;愈伤组织诱导最佳培养基为MS+2,4-D 1.0 mg/L+6-BA 0.1 mg/L;腋芽诱导最佳培养基为MS+6-BA 1.0 mg/L+NAA0.5 mg/L;愈伤组织增殖最佳培养基为MS+TDZ(0.2~1.0)mg/L+NAA(0.02~0.05)mg/L;愈伤组织分化最佳培养基为MS+6-BA(0.5~1.0)mg/L+NAA(0.02~0.05)mg/L;芽增殖最佳培养基为MS+6-BA 1.5 mg/L+NAA0.5 mg/L;最佳生根培养为1/2MS+IBA 1.0 mg/L+NAA 0.2 mg/L。试管苗在珍珠岩和腐殖质(1∶1)基质中炼苗,成活率达75%以上。结论:筛选出了外植体适宜的消毒方法及愈伤组织诱导、增殖、分化及芽增殖、生根的适宜培养基,建立了‘渝蕾一号’组培快繁体系。
OBJECTIVE: To establish a tissue culture rapid propagation system of Lonicera macranthoides Hand.-Mazz.Yulei1, a new species of Lonicera macranthoides, which provides a theoretical basis for its industrialized breeding. Methods: Young stem segments and leaves of ’Yule-1’ were used as explants and MS as basic medium. Different concentrations of NAA, 6-BA, TDZ, IBA and other plant growth regulators were added at different stages of tissue culture Material, callus induction, proliferation and differentiation, axillary bud stem induction, proliferation and rooting of the system for the study. Results: The optimum sterilization time of explants was 2 ~ 3 min for stems and 1 ~ 2 min for leaves. The best medium for callus induction was MS + 2,4-D 1.0 mg / L + 6- BA 0.1 mg / L. The best culture medium for axillary bud induction was MS + 6-BA 1.0 mg / L + NAA 0.5 mg / L. The best medium for callus induction was MS + TDZ (0.2-1.0) mg / L + NAA (0.02-0.05) mg / L. The optimal culture medium for callus differentiation was MS + 6-BA (0.5-1.0 mg / L) and NAA The best rooting culture was 1 / 2MS + IBA 1.0 mg / L + NAA 0.2 mg / L for MS + 6-BA 1.5 mg / L + NAA 0.5 mg / L. Test-tube seedlings perlite and humus (1: 1) matrix refining, survival rate of 75% or more. Conclusion: The proper method of disinfection of explants and suitable medium for callus induction, proliferation, differentiation, bud multiplication and rooting were screened out. The rapid tissue culture system of ’Yulei No.1’ was established.