研究青钱柳细胞悬浮培养体系的建立及三萜化合物的积累.结果表明,以青钱柳无菌幼叶为外植体,可诱导出愈伤组织.选择绿色、较为疏松的愈伤组织继续筛选和继代培养,可获得疏松易碎、生长旺盛、适于悬浮培养的优良愈伤组织.青钱柳细胞优化培养条件为:MS培养基、2,4-D 0.8 mg/L、NAA1.8 mg/L、6-BA 0.8 mg/L、KT 0.8 mg/L、蔗糖40 g/L,pH值6.0,100 mL三角瓶中装液量为40 mL,接种量为3 g(鲜质量),暗培养.该条件下悬浮细胞生长迅速,最大生长量(干质量)为15.56 g/L,总三萜产量达1 179.85 mg/L,熊果酸和齐墩果酸是其中主要的三萜化合物,产量分别达443.64 mg/L和159.00mg/L.青钱柳细胞悬浮培养可用于熊果酸和齐墩果酸生产.
In order to provide references for industrialized production of secondary metabolites such as triterpenoids by plant cell culture,the establishment ofCyclocarya paliuruscell suspension culture system and the accumulationof triterpenoids were studied.Results show that callus could be induced from sterile immature leaves.The crispgreen callus was further selected for subculture and suspension culture.Suspended cells grew very well.The in-creased dry weight of cells could reach a level of15.56 g/Lunder the following conditions: MS media containing2,4-D 0.8 mg/L,NAA1.8 mg/L,6-BA0.8 mg/L,KT0.8 mg/Land 4 % sucrose,pH=6.0,inoculumconcen-tration(fresh weight) of3 g/40 mL,filling volume of40 mL/100 mL,dark cultivation.The output of total triter-penoids was up to 1 179.85 mg/L,ursolic acid and oleanolic acid were the major compounds of total triterpenoids,and its output reached 443.64 mg/L and 159.00 mg/L under the optimal culture conditions.These results suggestthatCyclocarya paliuruscell suspension culture could be applied to producing ursolic acid and oleanolic acid.