He Yan, Zhu Yu-Qiu, Xiao Bo, Fu Shun-Hua, Si Jin-Ping
State Key Laboratory of Subtropical Silviculture,Zhejiang A&F University Lin'an 311300,China.
Chongqing Institute of Medicinal Plant Cultivation Chongqing 408435,China.
Zhongguo Zhong Yao Za Zhi. 2019 May;44(10):2032-2037. doi: 10.19540/j.cnki.cjcmm.20190301.010.
In order to accelerate the breeding of the excellent seedlings of Polygonatum cyrtonema,tissue culture system of P. cyrtonema was established through the comprehensive regulation of key factors( leaf age,leaf location,basic media and plant growth regulators) and cytological basis of callus formation and differentiation was analyzed through paraffin section. The results showed that the 30-day-old leaf base explanton medium MS+6-BA 1. 50 mg·L(-1)+2,4-D 0. 20 mg·L(-1) had the highest induction rate( 80. 00%). The callus was initiated from cells on leaf base epidermis and near cortex,formed by the differentiation of middle vascular bundle cells. The optimal medium for adventitious bud differentiation was MS+ 6-BA 4. 00 mg·L(-1)+ 2,4-D 0. 20 mg·L(-1) with the differentiation rate of90. 33%,and the average number of buds was 5. 16. The adventitious buds had two origin types: exogenous and endogenous origin,formed by callus proximal cells and callus internal meristemoid. The adventitious bud proliferation medium was screened by orthogonal design,which determined the optimum combination was MS+ 6-BA 2. 00 mg·L(-1)+NAA 0. 10 mg·L(-1) and MS+ 6-BA 2. 00 mg·L(-1)+NAA 0. 20 mg·L(-1). The tubers with three leaves were cut and inoculated in the medium 1/2 MS+IBA 2. 00 mg·L~(-1),showing the highest rooting rate of 94. 00%. The rooting seedlings transplanted into the peat-vermiculite( 1 ∶ 1) matrix grew healthy and the survival rate was over 85. 00%. This research provided a novel solution for large-scale cultivation of P. cyrtonema seedling.
为加速黄精优良种苗的繁育，通过对关键因素（叶龄、叶位、基本培养基和植物生长调节剂）的综合调控建立了黄精组织培养体系，并通过石蜡切片分析了愈伤组织形成和分化的细胞学基础。结果表明，30日龄叶基外植体培养基MS+6-BA 1.50 mg·L⁻¹+2,4-D 0.20 mg·L⁻¹的诱导率最高（80.00%）。愈伤组织由叶基表皮和近皮层细胞起始，由中维管束细胞分化形成。不定芽分化的最佳培养基为MS+6-BA 4.00 mg·L⁻¹+2,4-D 0.20 mg·L⁻¹，分化率为90.33%，平均芽数为5.16。不定芽有外起源和内起源两种类型，分别由愈伤组织近端细胞和愈伤组织内部类分生组织形成。通过正交设计筛选不定芽增殖培养基，确定最佳组合为MS+6-BA 2.00 mg·L⁻¹+NAA 0.10 mg·L⁻¹和MS+6-BA 2.00 mg·L⁻¹+NAA 0.20 mg·L⁻¹。将三叶块茎切割接种于1/2 MS+IBA 2.00 mg·L⁻¹培养基中，生根率最高，为94.00%。生根苗移栽到泥炭-蛭石（1∶1）基质中生长良好，成活率超过85.00%。本研究为黄精种苗的规模化栽培提供了新的解决方法。
