濒危药材金线莲愈伤组织诱导及分化成苗研究

[目的]建立有效的濒危药材金线莲的再生体系,为其工厂化生产奠定技术基础。[方法]选取福建金线莲的叶片、茎片、带节茎段、不定芽为外植体,建立无菌体系,而后以愈伤组织的诱导率为指标,采用正交设计优化不同外植体各阶段(愈伤组织的诱导和分化)培养基中激素的种类及浓度。[结果]在试验的福建金线莲4种外植体中,不定芽的愈伤组织诱导率最高,可达96.67%,其最适培养基配方为MS+0.5 mg/L NAA+4.0 mg/L BA+0.3 mg/L KT。在此培养基上,诱导的愈伤组织在分化培养基MS+0.5 mg/L NAA+2.0 mg/L BA和增殖培养基MS+0.2 mg/L NAA+2.0 mg/L BA上继代形成了无根试管苗。[结论]该研究建立的无菌繁殖体系,解决了前人所指出的金线莲愈伤组织诱导率极低的瓶颈问题,组培成苗的效率有较大提高。

Study on Callus Induction and Seeding Differentiation of the Rare Medicinal Species Anoectochilus roxburghii

[Objective] The research aimed to establish an effective recycling system for Anoectochilus roxburghii, and lay the technical foundation for its industrial production.[Method] As explant of leaf, stem, stem segment with node and adventitious bud of Anoectochilus roxburghii, the aseptic system was established, and then the induction rate of callus was used as an index to optimize the type and concentration of hormones in the culture medium of different explants (callus induction and differentiation) by orthogonal design.[Result]In the test of Anoectochilus roxburghii, four explants, the callus induction rate of adventitious bud was the highest, up to 96.67%, the optimum medium formula was MS+0.5 mg/L NAA+4.0 mg/L BA+0.3 mg/L KT.On the basis of the medium, the callus was subcultured on the differentiation medium(MS+0.5 mg/L NAA+2.0 mg/L BA) and the proliferation medium (MS+0.2 mg/L NAA+2.0 mg/L BA) to form a rootless test tube.[Conclusion]Asepsis reproductive system has been established, which solves the bottleneck problem that the predecessors pointed out that the callus induction rate of Anoectochilus roxburghii was very low,the efficiency of tissue culture seedlings have greatly improved.