A novel strategy to enhance terpenoids production using cambial meristematic cells of Tripterygium wilfordii Hook. f.

Abstract

Background 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. Results 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. Conclusions 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.

Establishment and triterpenoid production of Ocimum basilicum cambial meristematic cells

“…In Taxus cuspidata CMCs, paclitaxel yield was found to be eightfold higher in CMCs than in DDCs (Lee et al 2010). Tripterygium wilfordii CMCs also yielded at least twofold higher terpenoid contents than DDCs (Song et al 2019). Across most cultivations in this work, productivities of oleanolic acid were found to be lower than of ursolic acid, which is consistent with previous reports on O. basilicum (Pandey et al 2019).…”

“…1a). Abundance of small vacuoles and generally small cell size compared to DDCs from the same species are distinguishing features widely described for CMCs (Lee et al 2010;Song et al 2019;Zhou et al 2015).To confirm the observed structures as small vacuoles in CMCs, they were stained with neutral red, which penetrates the cell wall and membranes, accumulating in vacuoles (Fig. 1c).…”

“…1d, red box, interquartile range 17.60 µm, min 21.30 µm, max 133.893 µm). Although previous reports state that CMCs are smaller than DDCs (Lee et al 2010;Song et al 2019), their sizes were not comparatively quantified before.…”

“…These cells were described to grow at a faster rate, aggregate less and accumulate more product than dedifferentiated cells (DDCs) of the same plant (Ochoa-villarreal et al 2015). CMCs were described for Taxus cuspidata (Lee et al 2010), Catharanthus roseus (Moon et al 2015;Zhu et al 2018) and Tripterygium wilfordii (Song et al 2019) among others, but not yet for O. basilicum.…”

Food ingredients and food made with plant cell and tissue cultures: State‐of‐the art and future trends

“…They are embedded in meristems located at the tips of shoots, as well as roots, or are contained inside the vascular system and can grow faster as well as reach higher product concentrations than DDC‐based suspension cells [61]. The technology for the generation of CMCs is still in its infancy and has been reported for Taxus cuspidata [38], Catharanthus roseus [62, 63], Ginko biloba and Solanum lycopersicum [64], Tripterygium wilfordii [65] and Ocimum basilicum (Mehring A, University of Kaiserslautern, personal communication, 2019). Unhwa Corporation having secured the world`s first patent technology in plant stem cell isolation is the leading company in this field.…”