- 期刊
Enhanced Tanshinone Production in Hairy Roots of "Salvia Miltiorrhiza Bunge" Under the Influence of Plant Growth Regulators in Liquid Culture
植物生長調節劑對丹參毛狀根液態培養累積丹參酮之影響
摘要
丹參(Salvia miltiorrhiza Bunge) 所含之二萜類丹參酮化合物於現代藥理研究證實其對許多疾病具有良好的療效。本研究以丹參葉片為材料，建立毛狀根培養系統，並探討植物生長調節劑對丹參毛狀根生產3 種丹參酮(tanshinone-I, tanshinone-IIA, cryptotanshinone) 之影響。以農桿根群菌 (Agrobacteriumrhizogenes）BCRC15010 菌系感染丹參葉片培植體可誘導毛狀根形成率達78%。轉殖毛狀根之鑑定則利用農桿根群菌rolB 及rolC 基因序列為引子，進行PCR 反應並比對其基因片段序列後加以證實。誘導所得之丹參毛狀根於無光照條件下培養於B5 液體培養基中具有生長迅速的特性。丹參毛狀根培養在含不同植物生長調節劑 (auxins, cytokinins 與abscisic acid) 的培養基中均可增加丹參酮類產生；毛狀根培養在含1 mg/L ABA (abscisic acid) 或TDZ (thidiazuron) 培養基中時，其tanshinone-I 與cryptotanshinone含量分別是溫室植株根部含量的5 與7.5 倍；添加TDZ, ABA 或BA (benzyladenine) 於培養基中可提升cryptotanshinone 之含量，其值分別為市售藥材之6.3, 5.0 與3.75 倍。本研究結果顯示添加植物生長調節劑可作為提升丹參酮產量之有效策略。
並列摘要
Tanshinone constituents are the most potent diterpene diketones used to treat several diseases. We studied the influence of different plant growth regulators (PGR) on the growth and production of three tanshinone constituents using hairy root culture of Salvia miltiorrhiza Bunge. Leaves of the medicinally-important plant S. miltiorrhiza were infected with Agrobacterium rhizogenes strain BCRC15010, which induced hairy roots in 78% of the explants. The best line of hairy roots was established on B5 liquid medium under dark conditions. The transformed nature of the root lines was confirmed by polymerase chain reaction using ”rol” B and C gene-specific primers. Adding different PGRs i.e., auxins, cytokinins and abscisic acid (ABA), enhanced the production of cryptotanshinone, tanshinone I and IIA. Tanshinone I and cryptotanshinone accumulationwas 5- and 7.5-fold higher with the addition of 1.0 mg/l ABA and TDZ, respectively, compared toroot of greenhouse-grown plant. Cryptotanshinone content was 6.3-, 5.0- and 3.75-fold higher in hairy roots grown under the influence of TDZ, ABA and BA, respectively, compared to a commercial herb. The devisedprotocol can be used to enhance tanshinone production.
延伸閱讀
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