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发根农杆菌诱导人参产生发根及离体发根中人参皂甙含量的测定
引用本文:赵寿经,杨振堂,李昌禹,臧埔,申玉华. 发根农杆菌诱导人参产生发根及离体发根中人参皂甙含量的测定[J]. 吉林农业大学学报, 2001, 23(2): 57-63
作者姓名:赵寿经  杨振堂  李昌禹  臧埔  申玉华
作者单位:1. 中国农业科学院特产研究所,
2. 吉林省药品检验所,
基金项目:InternationalCooperativeFoundationofChineseAcademyofAgriculturalScience,The Research Development Pro gram of JilinProvince ( 2 0 0 0 0 2 0 3 2 )
摘    要:发根农杆菌A4菌株诱导人参(Panax ginseng C.A.Meyer)产生发根,诱导率31.6%,转化发根可在无任何激素的MS培养基上快速生长,且具有分枝性强、丛生、无向地性等特点。通过对液体培养条件的选择,发现人参发根在1/2MS液体培养基上(25℃、110r/min摇床上暗培养)获得最大生长速率,经14d培养,人参发根鲜重增加了7.97倍。利用高效液相色谱法测定发根中人参皂甙含量发现,发根系R9923中人参单体皂甙Rb1含量达到10.38mg/g,超过栽培六年生人参根中Rb1含量(6.45mg/g)。用高压纸电泳方法在人参发根中检测到农杆碱及甘露碱的存在。Southern点杂交证明:本研究获得的人参发根确已被A4菌株Ri质粒的T-DNA所转化,并被整合到人参发根DNA上。

关 键 词:人参发根 遗传转化 人参皂甙 液体培养 人参 含量测定 发根农杆菌
文章编号:1000-5684(2001)02-0057-07
修稿时间:2001-03-13

Formation of Ginseng Hairy Roots Mediated by Agrobacterium rhizogenes and Analysis of Ginsenosides in Hairy Roots In Vitro
ZHAO Shou-jing,YANG Zhen-tang,LI Chang-yu,ZANG Pu,SHEN Yu-hua. Formation of Ginseng Hairy Roots Mediated by Agrobacterium rhizogenes and Analysis of Ginsenosides in Hairy Roots In Vitro[J]. Journal of Jilin Agricultural University, 2001, 23(2): 57-63
Authors:ZHAO Shou-jing  YANG Zhen-tang  LI Chang-yu  ZANG Pu  SHEN Yu-hua
Affiliation:ZHAO Shou jing 1,YANG Zhen tang 1,LI Chang yu 1,ZANG Pu 1,SHEN Yu hua 2
Abstract:Hairy roots of ginseng (Panax ginseng C. A. Meyer) were induced by strain A4 of Agrobacterium rhizogenes and the induction rate was 31.6%. The transformed hairy roots grew rapidly on MS medium without phytohormones and were characterized by strongly branching, clustering and non-geotropism, etc. The hairy roots of ginseng achieved the highest growth rate on MS liquid medium (shaked at 110 r/min at 25℃ in darkness) and their fresh weight was increased by 7.97 times after culture for 14 days. Determined by HPLC, the content of ginsenoside-Rb1 in hairy root clone R9923 was found to be 10.38 mg/g, higher than that of 6-year-old cultivated ginseng (6.45mg/g). Agropine and mannopine were detected in the hairy roots of ginseng by the high-voltage paper electrophoresis. Southern dot blotting proved that the hairy roots of ginseng had been indeed transformed by the strain A4 and the T-DNA of Ri plasmid had been integrated into the DNA of hairy roots.
Keywords:Ginseng hairy roots  genetic transformation  A rhizogenes A 4  ginsenosides  liquid culture
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