无机盐对甘草愈伤组织生长及总黄酮含量的影响

Effects of inorganic salts on the growth and total flavonoid content of Glycyrrhiza uralensis callus

本研究以甘草无菌苗的下胚轴为供试材料,以MS+6-BA 0.5mg/L+2,4-D 0.5mg/L+NAA 0.5mg/L+KT 0.4mg/L+agar 8g/L+sugar 30g/L为基本培养基,利用组织培养技术并结合紫外分光光度法探究了MS培养基中5种不同无机盐浓度及各种成分之间的交互作用对甘草愈伤组织增长量、生理指标及次生代谢产物的影响。结果表明,适量增加KNO_3和MnSO_4·4H_2O的含量有利于甘草愈伤组织生物量的积累;随着MS培养基中CaCl_2·2H_2O浓度的增加,有利于可溶性糖和总黄酮含量的积累;浓度为2533mg/L KNO_3最有利于提高POD活性;KH_2PO_4浓度在170~226 mg/L范围内,愈伤组织生长量和总黄酮含量显著提高;MS培养基中MgSO_4·7H_2O有利于甘草愈伤组织的生长及总黄酮的合成。而愈伤组织的生长状况与总黄酮的合成是多种无机盐离子之间交互作用的结果,通过正交试验及极差分析得出最优营养元素组合为MgSO_4·7H_2O 493 mg/L+MnSO_4·4H_2O 7mg/L+KNO_32533mg/L+KH_2PO_4170mg/L+CaCl_2·2H_2O 733mg/L。

In this study,five kinds of inorganic salts were added to Murashige and Skoog（MS）culture medium to determine their effects on the growth rate,physiological indexes,and secondary metabolite contents of licorice（Glycyrrhiza uralensis）callus.The hypocotyledonary axis of sterile licorice seedlings was used as the experimental material.The culture medium consisted of MS＋6-benyzlaminopurine（0.5 mg/L）＋2,4-dichlorophenoxyacetic acid（0.5mg/L）＋naphthalene acetic acid（0.5mg/L）＋kinetin（0.4mg/L）＋agar（8g/L）＋sugar（30g/L）.Tissue culture and ultraviolet spectrophotometry methods were used to study the effects of inorganic salts at various concentrations in the culture medium on the growth and characteristics of licorice callus.Increasing amounts of KNO3 and MnSO4·4H2O in the medium increased the biomass of licorice callus.Increasing concentrations of CaCl2·2H2O in the medium led to increased accumulation of soluble sugars and total flavonoids.The highest peroxidase activity was in callus in medium containing 2533 mg/L KNO3.Callus growth and total flavonoid content increased significantly as the KH2PO4 concentration increased from 170mg/L to 226mg/L.The presence of MgSO4·7H2O in the MS medium promoted licorice callus growth and flavonoid synthesis.However,callus growth and total flavonoid synthesis were affected by the interactions among various inorganic salts.The results of an orthogonal test and range analysis indicated that the optimal combination of salts for growth and secondary product accumulation in licorice callus was as follows：MgSO4·7H2O 493mg/L;MnSO4·4H2O 7mg/L;KNO32533mg/L;KH2PO4170mg/L;and CaCl2·2H2O 733mg/L.