%0 Journal Article
%T Transformation of Protoplast-derived Colonies by Agrobactenum tumefaciens in Taxus vunnanensis<br>根癌农杆菌对云南红豆杉原生质体的转化
%A Luo Jianping
%A Jiang Shaotong
%A Pan Lijun
%A <br>罗建平
%A 姜绍通
%A 潘丽军
%J 世界科学技术-中医药现代化
%D 2003
%I 
%X In order to isolate cell lines with high paclitaxel productivity via molecular mutagenesis, the transformation of Taxus yunnanensis protoplasts by Agrobacterium tumefaciens was studied. Protoplasts were obtained from friable, light yellow callus of T. yunnanensis in enzyme solution. Callus in the exponential growth phase of 15-20 days after subculture produced the highest yield of viable protoplasts. When cultured in liquid medium composed of B5 salts, KM vitamin and organic components and 0.45 mol/L fructose and supplemented with 3.0 mg/L 2,4-D and 0.1 mg/L KT, protoplasts undenvent sustained divisions and formed cell colonies. Although A. tumefaciens failed to transform freshly isolated protoplasts and protoplasts with regenerated cell walls, transformants were obtained by co-cultivating protoplast-derived minicolonies consisting of 10 or more celis with the strain B6S3. The transformants were confirmed by opine analysis. The transformation frequency was about 5%. HPLC analysis showed that there was a significant difference in paclitaxel content among all transformants. The highest paclitasel content in transformants was found to be 0.076%, which was 6 times as high as in control callus. The transformant with high paclitaxel accumulation showed a lower cell growth as compared to its control. During subculture the transformant remained unchanged with respect to cell growth and paclitaxel accumulation. Although the paclitaxel accumulation in the transformants was not high enough to produce paclitaxel for commercial purposes, the method described in the present study would provide an opportunity to isolate cell lines with high paclitaxel productivity from mutagenized single cell culture via T-DNA insertion.
%K 云南红豆杉
%K 原生质体
%K 根癌农杆菌
%K 转化
%K 紫杉醇
%U http://www.alljournals.cn/get_abstract_url.aspx?pcid=A9DB1C13C87CE289EA38239A9433C9DC&cid=B5F35A1FC27C1FF7&jid=44257A35316DB0727BA6BC707B9EDAAB&aid=29C0764AAC1C331F3F712D965E6F7D04&yid=D43C4A19B2EE3C0A&vid=94C357A881DFC066&iid=38B194292C032A66&sid=2A3781E88AB1776F&eid=286FB2D22CF8D013&journal_id=1674-3849&journal_name=世界科学技术-中医药现代化&referenced_num=0&reference_num=0