【摘要】 目的 :建立人参毛状根转化体系。方法 :利用含有Ri质粒的发根农杆菌Agrobacteriumrhizogenes感染人参Panaxginseng进行毛状根诱导 ,利用PCR技术及TLC分析方法对毛状根进行鉴定。 结果 :首次从人参带叶幼茎处诱导出毛状根 ,用激素和AS(乙酰丁香酮 )处理可提高转化率并缩短转化时间。毛状根在无激素B5培养基上生长并失去向地性 ,月增长倍数可达 5 0倍 (鲜重 )。PCR分析证实农杆菌Ri质粒含有rolC序列 (5 6 4bp)的T DNA已整合到人参转化株的染色体组中 ,TLC分析证明T DNA特异的表达产物冠瘿碱在转化细胞中合成。人参毛状根中人参总皂苷含量为 2 .486 % (干重 ) ,而人参原药材总皂苷含量为 1.40 3% (干重 )。结论 :人参毛状根生长迅速 ,皂苷含量高于原药材 ,该体系的建立为利用生物技术工业化生产人参活性成分奠定了基础。更多还原

【Abstract】 Objective:To obtain the hairy root of Panax ginseng (HRPG). Method:HRPG was obtained by infecting the germ free seedling,cotyledon,leaf the blade,Petiole with Agrobacterium rhizogenes 15834 harbouring agropine type Ri plasmid. The transformation was proved by PCR and TLC. Result: HRPG grew rapidly on hormone free medium and showed typical phenotype of hairy root. The induced rate and time can be improved by adding exogenous hormone and AS(hydroxyacetosyringone). PCR analysis confirmed the integration of TL DNA, including 564bp rolC sequence in the HRPG . Opine analysis evidenced the integration and expression of TL DNA. The total saponin in HRPG (2.486%) was higher the than control(1.403% Panax ginseng ).Conclusion: HRPG grows rapidly and proves high in content of ginseng saponin. This culture system of HRPG will be useful for the production of active components in ginseng.更多还原