摘要
葛根是传统中药。利用发根土壤杆菌 (Agrobacteriumrhizogenes (Rikeretal.)Conn)R16 0 1转化野葛 (Puerarialobata (Willd .)Ohwi)、山葛 (P .lobatavar.montana)和三裂叶野葛 (P .phaseoloides (Roxb .)Benth .)离体叶片 ,在叶片表面直接形成毛状根。毛状根的诱导频率分别为 16 .6 %、16 .2 %和 2 6 .6 %。三种药用葛毛状根均表现出生长快、多分支、负向重力性的典型特征。毛状根离体培养均具有激素自主特性和抗卡那霉素特性 ,毛状根的冠瘿碱纸电泳、rol基因扩增及分子杂交实验均证实了其转基因性质。
Pueraria radix (the dried root of Pueraria plant) is known as a traditional Chinese drug. Hairy roots of Pueraria lobata (Willd.) Ohwi, P.lobata var. montana and P. phaseoloides (Roxb.) Benth. transformed by Agrobacterium rhizogenes (Riker et al .) Conn R1601 were developed directly from the surface of sterile leaves in vitro . The transformation frequency was 16.6%, 16.2% and 26.6%, respectively. All hairy roots in the three species displayed the typical phenotypes of rapid growth, highly branched and plagiotropism, and also exhibited hormone autotrophy and resistance to kanamycin.The genetic transformations were confirmed by opine paper electrophoretic analysis, rol gene PCR amplification and molecular hybridization.
基金
广东省重点科技攻关项目!(S990 40 1)&&
关键词
发根土壤杆菌
毛状根
遗传转化
药用葛
葛根素
Agrobacterium rhizogenes
hairy root
genetic transformation
medical Pueraria
