甘草组培苗的培养基优化及其有效成分分析

Tissue Culture Medium Optimization and Active Ingredient Analysis of Liquorice Tissue Culture Plantlets

为了探明用甘草种子无菌幼苗不同部位进行组培繁育的优化条件及其有效成分含量,利用组织培养技术结合高效液相色谱法和紫外分光光度法研究了甘草幼苗各部位进行组培的适宜外植体、组培苗的优化培养基及其有效成分。结果表明:1)适宜分蘖培养的外植体为生长4～7d的无菌幼苗子叶段,培养基为MS+0.2mg/L NAA+1.0mg/L 6-BA。2)以MS+3.0mg/L IBA+0.1mg/L KT培养基进行生根培养,其真叶段的效果最好,成苗率达79.33%,但相同培养基又以茎尖的生根效果最好,成苗率达94.67%。3)组培苗移栽以培养30d以上,蛭石和河沙为基质的效果较好,移栽成活率可达88%以上。4)组培苗中检测的8种甘草黄酮成分都低于甘草药材,但甘草组培苗中的总黄酮含量高于甘草药材,多糠含量为154.2mg/g,极显著高于甘草药材(145.5mg/g)。甘草不同部位、培养基、移栽时间和基质都会显著影响甘草的成苗和移栽成活率,甘草组培苗都含有甘草黄酮和多糖等有效成分。

In order to explore the optimization conditions for tissue culture and breeding of different parts of sterile liquorice seedlings and their active ingredient contents,the suitable explants,optimum culture medium of tissue culture seedling and active ingredients of different parts of liquorice were studied by using tissue culture technique,HPLC and ultraviolet spectrophotometry.Results： 1） the suitable explant for tillering culture was 4~7 day-old sterile cotyledon segments and the culture medium was MS＋0.2 mg/L NAA＋1.0 mg/L 6-BA.2） the seedling rate of euphylla segment reached 79.33% on the culture medium MS＋3.0 mg/L IBA ＋0.1 mg/L KT,but the rooting effect of stem tip was the best on the same culture medium,the seedling rate was up to 94.67%.3） the transplanting survival rate of 30 day-age plantlets was over 88% when taking vermiculite and river sand as the substrates.4） the contents of the eight kinds of licorice flavonoids were all lower than that in liquorice medicinal material,but the total flavonoid content was higher than liquorice medicinal material;the polysaccharide content in liquorice tissue culture plantlet was 154.2 mg/g,significantly higher than that in liquorice medicinal material（145.5 mg/g）.Various parts of liquorice,culture medium,transplanting time and substrate could all significantly influence the seedling rate and transplanting survival rate.All the liquorice tissue culture plantlets contained liquorice flavonoids and liquorice polysaccharide.