| Ri质粒人参转化系统的建立及鉴定 |
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| 引用本文： | 刘峻,丁家宜,徐红,王建波.Ri质粒人参转化系统的建立及鉴定[J].中国中药杂志,2001,26(2):95-99. |
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| 作者姓名： | 刘峻 丁家宜 徐红 王建波 |
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| 作者单位： | 中国药科大学 中药学院 生物技术研究室, |
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| 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Agrobacterium rhizogenes感染人参Panax ginseng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 50倍 (鲜重 )。PCR分析证实农杆菌Ri质粒含有rolC序列 (564bp)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 1.403% (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。
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| 关 键 词： | 发根农杆菌 人参毛状根 聚合酶链式反应(PCR) 冠瘿碱 |
| 文章编号： | 1001-5302(2001)02-095-05 |
| 收稿时间： | 2000/5/30 0:00:00 |
| 修稿时间： | 2000年5月30日 |
Genetic Transformation of Panax gingseng C.A. Meyer Induced by Root Inducing Plasmid(Ri) of Agrobacterium rhizogenes |
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| LIU Jun;DING Jia yi;WANG Jiang bo.Genetic Transformation of Panax gingseng C.A. Meyer Induced by Root Inducing Plasmid(Ri) of Agrobacterium rhizogenes[J].China Journal of Chinese Materia Medica,2001,26(2):95-99. |
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| Authors: | LIU Jun;DING Jia yi;WANG Jiang bo |
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| Affiliation: | Department of Biotechnology, China Pharmaceutical University, Nanjing 210038, Jiangsu, China. |
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| 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. |
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| Keywords: | Agrobacterium rhizogenes hairy roots of Panax ginseng (HRPG) PCR opine |
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