组织培养和HPLC法研究红豆杉生物合成紫杉醇的组织化学定位

Study on the Histochemical Localization of Taxus Plants Biosynthesis Taxol by Tissue Culture and HPLC

为了探索用组织培养法和高效液相色谱(HPLC)法研究红豆杉植物生物合成紫杉醇的组织化学定位,对南方红豆杉[Taxuschinensis(Pilger)Rehd.var.mairei(LemeeetL啨vl.)ChengetL.K.Fu]和曼地亚红豆杉(Taxusmediacv.Hicksii)各种组织进行了愈伤组织诱导培养,用HPLC法(检测条件:色谱柱为DiamonsilTMC18,5μm,4.60×250mm柱,检测波长为227nm,流动相为甲醇∶水∶乙腈=1∶2∶2,流速为1.2mlmin,柱温为35℃,灵敏度为0.16AuFs,进样量10μL)对组织培养物中紫杉醇含量进行检测,结果显示:红豆杉幼茎、幼茎皮、茎形成层、幼叶和顶端分生组织的组织培养物中紫杉醇含多以由分生组织细胞诱导培养所形成的愈伤组织为高。南方红豆杉以幼茎皮和茎形成层愈伤组织培养物中为高,分别为0.0409%(干重)和0.0407%(干重),曼地亚红豆杉则以茎形成层和顶端分生组织的愈伤组织培养物中为高,分别为0.025%(干重)和0.016%(干重),均是它们天然材料中含量的2.3～8倍。

For exploring the histochemical localization of Taxus plants biosynthesis taxol by tissue culture and HPLC, the various tissues of Taxus chinensis （Pilger） Rehd. vat. mairei（Le mee et Levl. ） Cheng et L. K. Fu and T. media cv. Hicksii were induced to product callus, then taxol content of the callus cultures was analysed by I-IPLC（The measure condition： Diamonsil^TM C18,5μm,4.60 × 250mm column, at 227nm wavelength is measured by ultraviolet dectector, the compose of flow phase is methanol： water： acetouitrile = 1 ： 2： 2, the velocity of flow is 1.2mL/min, the column temperature is 35%, the sensitivity is 0.16 AuFs） .The result shows： taxol content in the the young stems, the bark of the young stems,the stem cambium,the immature needle and the apical meristem callus cultures,the most of higher content callus cultures are meristem callus cultures, taxol content in the the young stems bark and the stem cambium callus cultures of T. mairei is higher, is 0.0409 % （DW） and 0.0407 % （DW）, and that in T. media stem cambium and the apical meristem callus cultures is higher, is 0.025% （DW） and 0.016% （DW）, the all of them is higher than 2.3 - 8times content of them natural material.