Tripterygium wilfordii Hook. f. (T. wilfordii) is an important medicinal plant with anti-inflammatory, immunosuppressive and anti-tumor activities. The main bioactive ingredients are diterpenoids and triterpenoids, such as triptolide, triptophenolide and celastrol. However, the production of terpenoids from original plants, hairy roots and dedifferentiated cells (DDCs) are not satisfactory for clinical applications. To find a new way to further improve the production of terpenoids, we established a new culture system of cambial meristematic cells (CMCs) with stem cell-like properties, which had strong vigor and high efficiency to produce large amounts of terpenoids of T. wilfordii. CMCs of T. wilfordii were isolated and cultured for the first time. CMCs were characterized consistent with stem cell identities based on their physiological and molecular analysis, including morphology of CMCs, hypersensitivity to zeocin, thin cell wall and orthogonal partial least square-discriminant analysis, combination of transcriptional data analysis. After induction with methyl jasmonate (MJ), the maximal production of triptolide, celastrol and triptophenolide in CMCs was 312%, 400% and 327% higher than that of control group, respectively. As for medium, MJ-induced CMCs secreted 231% triptolide and 130% triptophenolide at the maximum level into medium higher than that of control group. Maximal celastrol production of induced CMCs medium was 48% lower than that of control group. Long-term induction significantly enhanced the production of terpenoids both in cells and medium. The reason for increasing the yield of terpenoids was that expression levels of 1-deoxy-d-xylulose-5-phosphate synthase (DXS), 1-deoxy-d-xylulose-5-phosphate reductoisomerase (DXR) and hydroxymethylglutaryl-CoA synthase (HMGS) were upregulated in CMCs after induction. For the first time, CMCs of T. wilfordii were isolated, cultured, characterized and applied. Considering the significant enrichment of terpenoids in CMCs of T. wilfordii, CMCs could provide an efficient and controllable platform for sustainable production of terpenoids, which can be a better choice than DDCs.

中文翻译：

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使用雷公藤钩的形成层分生细胞增强萜类化合物生产的新策略。F。

雷公藤钩。F。(T. wilfordii) 是一种重要的药用植物，具有抗炎、免疫抑制和抗肿瘤活性。主要的生物活性成分是二萜类和三萜类，如雷公藤内酯、雷公藤酚和天竺葵素。然而，从原始植物、毛状根和去分化细胞（DDCs）生产萜类化合物的临床应用并不令人满意。为了寻找进一步提高萜类化合物产量的新途径，我们建立了具有干细胞特性的形成层分生细胞（CMCs）培养体系，该体系具有强大的活力和高效生产大量的T. wilfordii萜类化合物。 . T. wilfordii 的CMCs 首次被分离和培养。基于其生理和分子分析，包括CMC的形态、对zeocin的超敏性、薄细胞壁和正交偏最小二乘判别分析、转录数据分析的组合，CMC的特征与干细胞特性一致。用茉莉酸甲酯（MJ）诱导后，CMCs中雷公藤内酯、紫杉醇和雷公藤酚的最大产量分别比对照组高312%、400%和327%。培养基方面，MJ诱导的CMCs分泌的雷公藤内酯含量为231%，雷公藤内酯含量为130%，高于对照组。诱导 CMCs 培养基的最大 celastrol 产量比对照组低 48%。长期诱导显着增强了细胞和培养基中萜类化合物的产生。萜类化合物产量增加的原因是1-脱氧-d-木酮糖-5-磷酸合酶（DXS）、1-脱氧-d-木酮糖-5-磷酸还原异构酶（DXR）和羟甲基戊二酰辅酶A合酶的表达水平提高。 HMGS）在诱导后在 CMC 中上调。首次分离、培养、表征和应用了 T. wilfordii 的 CMC。考虑到T. wilfordii CMCs中萜类化合物的显着富集，CMCs可以为萜类化合物的可持续生产提供一个高效可控的平台，是比DDCs更好的选择。wilfordii 被分离、培养、表征和应用。考虑到T. wilfordii CMCs中萜类化合物的显着富集，CMCs可以为萜类化合物的可持续生产提供一个高效可控的平台，是比DDCs更好的选择。wilfordii 被分离、培养、表征和应用。考虑到T. wilfordii CMCs中萜类化合物的显着富集，CMCs可以为萜类化合物的可持续生产提供一个高效可控的平台，是比DDCs更好的选择。