多花黄精组织培养体系的研究

Study on tissue culture system of Polygonatum cyrtonema

为加快多花黄精优良种苗的繁育,通过叶片叶龄及部位、基本培养基和植物生长调节剂等关键因子的综合调控,建立多花黄精的组培快繁体系并通过细胞学观察解析愈伤组织形成与分化。结果表明:30 d龄多花黄精试管苗叶基部在MS+6-BA 1. 50 mg·L^-1+2,4-D 0. 20 mg·L^-1培养基上愈伤组织诱导率最高,为80. 00%,愈伤组织脱分化启动及愈伤组织的形成是多起源的;不定芽分化最佳培养基为MS+6-BA 4. 00 mg·L^-1+2,4-D 0. 20 mg·L^-1,分化率达90. 33%,每块愈伤组织平均分化芽数5. 16个,不定芽有外起源和内起源2种发生方式,分别由愈伤组织近表层细胞和愈伤组织内部拟分生组织形成;采用正交试验设计筛选出不定芽最佳增殖培养基为MS+6-BA 2. 00 mg·L^-1+NAA 0. 10 mg·L^-1和MS+6-BA 2. 00 mg·L^-1+NAA0. 20 mg·L^-1;切分三叶无菌苗接入培养基1/2 MS+IBA 2. 00 mg·L^-1中,生根率最高,为94. 00%;移栽于泥炭-蛭石1∶1的基质中生长良好,存活率达85. 00%以上。该研究为多花黄精工厂化育苗提供了关键解决途径。

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 of 90. 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.