PRODUCTION OF TROPANE ALKALOIDS BY HAIRY ROOT CULTURES OF DUBOISIA-LEICHHARDTII TRANSFORMED BY AGROBACTERIUM-RHIZOGENES
PRODUCTION OF TROPANE ALKALOIDS BY HAIRY ROOT CULTURES OF DUBOISIA-LEICHHARDTII TRANSFORMED BY AGROBACTERIUM-RHIZOGENES
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DOI：
10.1016/0168-9452(89)90137-4
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发表时间：
1989-01-01
期刊：
影响因子：
5.2
通讯作者：
YAMADA, Y
中科院分区：
文献类型：
--
作者：
MANO, Y;OHKAWA, H;YAMADA, Y
通过用发根农杆菌菌株 15834 和 A4 接种 Duboisia leichhardtii F. Muell 的叶盘和茎段来诱导毛状根。通过在不含植物激素的 HF（改良 Heller's）培养基中培养毛状根至少 5 代，建立了源自单个根分生组织的 45 个毛状根克隆。已建立的毛状根克隆的表征表明，不同克隆的生长速率、生物碱含量和生产力存在相当大的差异。产生最多东莨菪碱的克隆DL-34是通过选择多个毛状根克隆而建立的。 DL-34 在 HF 培养基中连续传代 12 代后，快速生长并伴有广泛的侧枝、冠脒产量和高东莨菪碱产量等特性均保持稳定；因此，检查了该克隆的生长和生物碱生产的培养条件。 25%避光培养4周后，其重量增加了64倍，在各最佳培养基中以干重计东莨菪碱含量为1.8%。该含量是 D. leichhardtii 植物叶子含量的 2 倍。培养 4 周后，克隆 DL-34 产生 78 mg/l 的东莨菪碱。我们表明，选择和培养基组成是获得良好生长和东莨菪碱产量的重要因素。
Hairy roots were induced by inoculating leaf disks and stem segments of Duboisia leichhardtii F. Muell with Agrobacterium rhizogenes strains 15834 and A4. Forty-five hairy root clones that had been derived from individual root meristems were established by culturing hairy roots for at least 5 passages in phytohormone-free HF (modified Heller''s) medium. Characterization of the established hairy root clones showed that there was considerable variation in growth rate, alkaloid content and productivity from clone to clone. Clone DL-34, which produced the most scopolamine, was established from the selection of a number of hairy root clones. Properties such as rapid growth accompanied by extensive lateral branching, opine production and high scopolamine production were stable in DL-34 for 12 successive passages in HF medium; therefore, culture conditions for growth and alkaloid production were examined for this clone. After 4 weeks of culture at 25% in the dark, its weight had increased 64-fold, and its scopolamine content was 1.8% on a dry weight basis in each optimum medium. This content was 2-fold that of the leaves of D. leichhardtii plants. Clone DL-34 produced 78 mg/l of scopolamine after 4 weeks of culture. We show that selection and medium-composition are essential factors in obtaining good growth and scopolamine production.