南方红豆杉愈伤组织培养及其紫杉烷二萜类成分的分析  

Tissue culture and analysis on taxane diterpenoids in callus of Taxus chinensis var. mairei

作　　者：姚晓[1] 步达[1] 陈建伟[1,2] 李祥[1,3] 汤彬[1] 

机构地区：[1]南京中医药大学药学院,江苏南京210023 [2]江苏省方剂研究重点实验室,江苏南京210023 [3]江苏省中药炮制重点实验室,江苏南京210023

出　　处：《中草药》2014年第18期2696-2702,共7页Chinese Traditional and Herbal Drugs

基　　金：江苏高校优势学科建设工程资助项目(YSXK2010);中药学优势学科开放课题(2011ZYX6-008)

摘　　要：目的研究南方红豆杉愈伤组织的出愈时间、诱导率、生长率、生长状态、褐变程度,比较愈伤组织中紫杉醇等5种紫杉烷二萜类成分的量。方法采用单因素比较、正交试验设计,研究影响南方红豆杉愈伤组织诱导和生长的各种因素;采用HPLC法测定不同来源愈伤组织中10-脱乙酰基巴卡丁Ⅲ(10-DAB)、巴卡亭Ⅲ、三尖杉宁碱、紫杉醇和东北红豆杉素5种紫杉烷二萜类成分的量;采用加权评分综合考虑以上各因素优选组织培养条件。结果外植体脱分化形成愈伤组织的能力,以完整叶片及茎段最好;暗培养有利于紫杉醇等5种萜类成分的富集;愈伤组织诱导的最适培养基为B5+2,4-D 3.0mg/L+6-BA 1.0 mg/L+NAA 1.0 mg/L+KT 0.5 mg/L。结论南方红豆杉愈伤组织诱导、培养与外植体种类、培养基种类、光照有较大关联,紫杉醇等5种紫杉烷二萜类成分的富集受植物生长调节剂种类及浓度影响显著。Objective To investigate some factors which could influence the tissue culture of callus from Taxus chinensis var. mairei such as the culturing callus time, callus induction rate, callus development rate, growth rate, and browning degree. Then, the five taxane diterpenoids including paclitaxel in callus were studied and compared. Methods The factors which influence the T. chinensis var. mairei callus induction and growth rate were studied by single factor analysis and orthogonal test design. The five taxane diterpenoids including paclitaxel in callus from different sources were determined by HPLC analysis. The weighted score for each of the above factors mentioned would be taken account to optimise the tissue culture conditions. Results Among the explants, the stems intacted leaves would exhibit the best ability of dedifferentiation to form callus. The enrichment dark culture was conducive to paclitaxel and other five kinds of terpene constituents. The most optimum culture medium of the callus induction was B5 ~ 2, 4-D 3.0 mg/L ＋ 6-BA 1.0 mg/L ＋ NAA 1.0 mg/L ＋ KT 0.5 mg/L. Conclusion The callus culture conditions for T. chinensis vat. mairei have indicated greater association with different types of the explantation, nutrient medium, and light illumination. The five kinds of enrichment taxane paclitaxel diterpenoids would be significantly affected by the type and concentration of the plant growth regulators. This study has shown some references value for T. chinensis var. mairei tissue culture and for the screening of high-yielding cell lines.

关 键 词：南方红豆杉 愈伤组织 加权评分 紫杉醇 紫杉烷二萜类 

分 类 号：R931.2[医药卫生—生药学]