- 期刊
何首烏毛狀根誘導與Emodin及Physcion之成分分析
Agrobacterium rhizogenes-mediated Transformation of Polygonum multiflorum THUNBERG and Quantitative Analysis of Emodin and Physcion
摘要
本試驗之目的為建立何首烏毛狀根轉殖系統與毛狀根轉殖株的鑑定方法，並以emodin及physcion為指標成分，進行何首烏的市售品、栽培的塊根及毛狀根成分含量比較，以探討利用毛狀根培養方式生產植物二次代謝物之可行性。結果發現毛狀根的誘導，以農桿根群菌BCRC15722的菌種感染何首烏節間、葉片及葉柄培植體時，以葉片誘導發根的效果最佳（約32%）。而在毛狀根的鑑定方面，以農桿根群菌的rol B基因與rol C基因，可應用於鑑定所誘導產生的毛狀根系，是否為轉殖成功的毛狀根。高效液相層析儀(HPLC)之分析測得田間何首烏塊根emodin(0.19 mg g^(-1))及physcion(0.15 mg g^(-1))的含量均高於市售品的含量（分別為emodin 0.08 mg g^(-1)及physcion 0.08 mg g^(-1)），但何首烏毛狀根則沒有檢測到emodin及physcion的累積。
並列摘要
The main objectives of the present investigation were (1) to use Agrobacterium rhizogenes mediated transformation to establish a hairy root culture and detection of rol genes in transgenic tissue of Polygonum multiflorum THUNBERG; (2) to analyze and compare the amount of emodin and physcion present in the crude drugs (processed underground or stem parts of P. multiflorum), field grown plants (underground parts) and in vitro grown hairy root by HPLC; and (3) to establish methods for the rapid production of higher amounts of secondary metabolite materials using hairy root culture technology. Of was found that thirty-two percentage of the leaf pieces induced hairy roots after co-culturing with Agrobacterium rhizogenes BCRC15722. The rol B and rol C genes in the hairy roots were confirmed by PCR analysis. HPLC analysis revealed that the major medicinal compounds-emodin and physcion in the field grown plants (emodin 0.19 mg g^(-1) and physcion 0.15 mg g^(-1)) were higher than those of the marketed crude drug (emodin 0.08 mg g^(-1) and physcion 0.08 mg g^(-1)). But, hairy root did not accumulate emodin and physcion.