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  

Formation of Ginseng Hairy Roots Mediated by Agrobacterium rhizogenes and Analysis of Ginsenosides in Hairy Roots In Vitro

作　　者：赵寿经[1] 杨振堂[1] 李昌禹[1] 臧埔 申玉华[2] 

机构地区：[1]中国农业科学院特产研究所,吉林吉林132109 [2]吉林省药品检验所,吉林长春130062

出　　处：《吉林农业大学学报》2001年第2期57-63,共7页Journal of Jilin Agricultural University

基　　金：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 (PanaxginsengC .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 ( 2 5℃、1 1 0r/min摇床上暗培养 )获得最大生长速率 ,经 1 4d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R992 3中人参单体皂甙Rb1 含量达到 1 0 .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。Hairy roots of ginseng ( Panax ginseng C A Meyer) were induced by strain A 4 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 Rb 1 in hairy root clone R9923 was found to be 10 38 mg/g, higher than that of 6 year old cultivated ginseng (6 45?mg/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 A 4 and the T DNA of Ri plasmid had been integrated into the DNA of hairy roots

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 

分 类 号：S567.51[农业科学—中草药栽培]