论文部分内容阅读
目的 :建立菘蓝 ( Isatis indigotica Fort)的毛状根离体培养系统 ,并诱导毛状根再生植株。 方法 :分别用发根农杆菌A4,R16 0 1和 ATCC15 834三种菌株感染菘蓝的子叶外植体 ,获得毛状根 ,并筛选了优质株系 ;测定了毛状根的生长曲线 ;在含不同激素的培养基上诱导毛状根再生植株 ;利用高压纸电泳法对毛状根和再生植株进行 T- DNA转化的检测。 结果 :首次利用发根农杆菌 A4,R16 0 1和 ATCC15 834三种菌株成功地从菘蓝中诱导出毛状根 ;菘蓝毛状根在含 BA的 MS培养基中成功地诱导出再生植株 ;经高压纸电泳检测 ,菘蓝毛状根及其再生植株中均含有甘露碱 ,表明 Ri质粒的 T- DNA已整合进毛状根和再生植株中。 结论 :菘蓝毛状根离体培养和植株再生系统的建立 ,为进一步进行药用活性成分的工业化生产和引入外源基因改良性状奠定了基础。
Objective : To establish a hairy root explant culture system of Isatis indigotica Fort and induce hairy root regeneration. METHODS: The cotyledon explants of A. indica were infected with Agrobacterium rhizogenes A4, R16 01 and ATCC15 834, respectively, to obtain hairy roots. High-quality strains were selected and the growth curve of hairy roots was determined. Hairy root regeneration plants were induced on media containing different hormones; hairy roots and regenerated plants were examined for T-DNA transformation by high pressure paper electrophoresis. Results: Agrobacterium rhizogenes A4, R16 0 1 and ATCC 15 834 were used for the first time to induce hairy roots from indigo. Indigo hairy roots were successfully induced in MS medium containing BA. The high-yield paper electrophoresis showed that the hairy roots and their regenerated plants contained mannopine, indicating that the Ri plasmid T-DNA had been integrated into hairy roots and regenerated plants. Conclusion : The in vitro culture of indigo hairy roots and the establishment of plant regeneration system lay the foundation for the further industrial production of medicinal active ingredients and the introduction of exogenous genetic improvement traits.