侧柏茎段培养快速繁殖技术研究和黄帝手植柏克隆

以3个不同年龄段的侧柏茎段为外植体，探讨基本培养基、植物生长调节剂、大量元素对侧柏茎段培养快繁的影响，研究侧柏茎段培养快速繁殖技术，并分析年龄对培养各阶段的影响，进而建立了黄帝手植柏克隆繁殖技术体系。结果表明，在试验范围内，各年龄段的侧柏茎段消毒效果均随消毒时间延长而提升，黄帝手植柏最佳升汞消毒时间为8 min；各年龄段的侧柏外植体在不同培养基上启动率均大于90%，选择MS+0.1 mg·L^-1 NAA+0.1 mg·L^-1 6-BA作为黄帝手植柏适宜启动培养基，启动率可达100%；50 a侧柏适宜在较高激素浓度的增殖培养基上培养，800、3 000 a侧柏适宜在中等激素水平的培养基上培养，黄帝手植柏适宜增殖培养基为MS+0.1 mg·L^-1 NAA+0.2 mg·L^-1 6-BA，30 d增殖系数为2.52；随着侧柏年龄增大，茎段生根更加困难，800、3 000 a侧柏在培养基MS+0.1 mg·L^-1 IBA+0.1 mg·L^-1 6-BA+0.1 mg·L^-1 NAA上有较好的生根效果，黄帝手植柏适宜生根培养基为MS+0.1 mg·L^-1 IBA+0.1 mg·L^-1 6-BA+0.1 mg·L^-1 NAA。本研究初步建立了黄帝手植柏茎段培养技术体系，为黄帝手植柏的克隆和种质资源保存奠定了技术基础。

Rapid Propagation of Platycladus orientalis by Stem Segment Culture and Clone of the P.orientalis Tree Planted by Huangdi

Stem segments of three different-aged Platycladus orientalis trees were used as explants to study the effects of basic medium,hormone and macroelement on stem segment culture.The rapid propagation technology was studied,and the influences of tree age on each stage of culture were analyzed.Furthermore,the clonal propagation technology of the P.orientalis tree planted by Huangdi was established.The results showed that under the conditions of the test,the disinfection effect of all aged segments was improved with the prolongation of disinfection time,and the optimal disinfection time was 8 min.The activation rate of all the explants on different media reached more than 90%.MS+0.1 mg·L^-1 NAA+0.1 mg·L^-16-BA was selected as the suitable starting medium,and the starting rate could reach 100%.Segments of 50-year-old tree was suitable for culturing on the proliferation medium with higher hormone concentration,while the segments of 800-and 3000-year-old tree were suitable for culturing on the medium with moderate hormone-level.The optimal medium for proliferation culture of the P.orientalis tree planted by Huangdi was MS+0.1 mg·L^-1 NAA+0.2 mg·L^-16-BA,from which the proliferation factor was 2.52 in 30 days.With the increase of the tree age,the segments were more difficult to root.The rooting effects of 800-and 3000-year-old tree were better on the medium MS+0.1 mg·L^-1 IBA+0.1 mg·L^-16-BA+0.1 mg·L^-1 NAA,which was also the optimal medium for rooting of the P.orientalis tree planted by Huangdi.The technical system for culturing the segment of the P.orientalis tree planted by Huangdi was initially established,which laid a technical foundation for the cloning and germplasm resource preservation of this most precious tree in China.