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[目的]研究野生中药资源血叶兰的无菌播种技术,实现其快速繁殖。[方法]以血叶兰种子为外植体,接种于4种不同的培养基MS、1/2MS、花宝1号(3.0 g/L)和1/2花宝1号(1.5g/L),80 d后统计萌发率,筛选适合血叶兰种子萌发的培养基。在培养基中添加不同激素配比,不同的有机物等,筛选适宜血叶兰原球茎增殖与分化、壮苗与生根的培养基。[结果]成熟血叶兰种子在1/2花宝1号(1.5 g/L)培养基中暗培养80 d左右形成原球茎团,萌发率为86.67%,是最适于血叶兰种子萌发的培养基;而种子萌发效果较差的是MS培养基。在1/2花宝1号(1.5 g/L)+6-BA 2.0 mg/L+椰汁100ml/L培养基中添加NAA 0.4-0.6 mg/L,原球茎团转接培养45 d左右形成原球茎和小芽的混合体,最高增殖倍数为6.11倍,增殖效果显著优于添加NAA 0.2和0.8 mg/L的培养基(P<0.05)。将培养获得的小芽转接至分别添加10%土豆汁、10%香蕉汁和100 ml/L椰汁的花宝1号(3.0 g/L)+NAA 1.0 mg/L+培养基中进行壮苗和生根培养,结果表明添加香蕉汁10%的培养效果最佳,培养80 d后可形成完整植株,幼苗生根率为93.33%,株高7.19 cm。经温室大棚炼苗10 d后,将血叶兰试管苗移栽至泥炭土与1 cm火山石(3∶1)的混合基质中,60 d后成活率达91.7%。[结论]该研究以血叶兰种子为外植体所建立的血叶兰无菌播种育苗技术体系可用于血叶兰的种质资源保护、种苗生产及产业化开发等。
[Objective] The research aimed to study the aseptic sowing technology of the wild Chinese medicinal materials, such as the blood-leaf orchid, to realize its rapid propagation. [Method] The seeds of Heteromonas flavescens were used as explants and inoculated into four different media MS, 1 / 2MS, Hua Bao 1 (3.0 g / L) and 1/2 Huabao 1 (1.5 g / L ). After 80 days, the germination rate was calculated and the suitable medium for the seed germination of P. septempunctata was screened. In the medium to add different hormone ratio, different organic matter, screening appropriate Heterophalranthes bulbi proliferation and differentiation, strong seedling and rooting medium. [Result] The mature Pueraria lobata seedlings formed protocorm groups in the medium of 1/2 Huabao 1 (1.5 g / L) for about 80 days with the germination rate of 86.67%, which was the most suitable for the seed germination Of the medium; and poor seed germination MS medium. The NAA 0.4-0.6 mg / L was added into the medium of 1/2 Huabao 1 (1.5 g / L) + 6-BA 2.0 mg / L + coconut milk 100 ml / L, and the protocorm groups were transferred and cultured for about 45 days The combination of bulbs and shoots had a maximum multiplication factor of 6.11, which was significantly better than that of NAA 0.2 and 0.8 mg / L (P <0.05). The small shoots were transferred to the medium of Huabao 1 (3.0 g / L) + NAA 1.0 mg / L + with 10% potato juice, 10% banana juice and 100 ml / And rooting. The results showed that 10% of banana juice was best cultured. After 80 days, the whole plant could be formed. The rooting rate of seedling was 93.33% and plant height was 7.19 cm. After incubation for 10 d in the greenhouse, the test tube was transplanted into a mixed matrix of peat soil and 1 cm volcanic stone (3: 1). The survival rate reached 91.7% after 60 days. [Conclusion] The system of seeding and nursery of sepia was established in this study using the sepia sepium as explant, which can be used for germplasm conservation, seedling production and industrialization development of geranium.