Establishment and triterpenoid production of Ocimum basilicum cambial meristematic cells

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ORIGINAL ARTICLE

Establishment and triterpenoid production of Ocimum basilicum cambial meristematic cells Alexander Mehring1 · Janik Haffelder1 · Jonas Chodorski1 · Judith Stiefelmaier1 · Dorina Strieth1 · Roland Ulber1 Received: 10 June 2020 / Accepted: 1 October 2020 © The Author(s) 2020

Abstract The application of plant suspension culture to produce valuable compounds, such as the triterpenoids oleanolic acid and ursolic acid, is a well-established alternative to the cultivation of whole plants. Cambial meristematic cells (CMCs) are a growing field of research, often showing superior cultivation properties compared to their dedifferentiated cell (DDC) counterparts. In this work, the first-time establishment of O. basilicum CMCs is demonstrated. DDCs and CMCs were cultivated in shake flasks and wave-mixed disposable bioreactors (wDBRs) and evaluated regarding triterpenoid productivity and biomass accumulation. CMCs showed characteristic small vacuoles and were found to be significantly smaller than DDCs. Productivities of oleanolic and ursolic acid of CMCs were determined at 3.02 ± 0.76 mg/(l*d) and 4.79 ± 0.48 mg/ (l*d) after 19 days wDBR cultivation, respectively. These values were consistently higher than any productivities determined for DDCs over the observed cultivation period of 37 days. Elicitation with methyl jasmonate of DDCs and CMCs in shake flasks resulted in increased product contents up to 48 h after elicitor addition, with the highest increase found in CMCs at 232.30 ± 19.33% (oleanolic acid) and 192.44 ± 18.23% (ursolic acid) after 48 h. Key message  For the first time, cambial meristematic cells of Ocimum basilicum were established and cultivated in a disposable bioreactor system. These cells outperform dedifferentiated cells of the same organism regarding productivity. Keywords  Plant cell culture · Cambial meristematic cells · Triterpenoids · Disposable bioreactor · Elicitation

Introduction The perennial herb Ocimum basilicum (family Lamiaceae), commonly known as basil, is valued worldwide for its culinary uses, but its applications go well beyond that. Basil and its essential oil are often used in traditional medicine (Irondi et al. 2016; Ezeani et al. 2017; Bae et al. 2020), cosmetics (Vivas Castaño et al. 2016; Volpe et al. 2018; Yeşil et al. 2020), and Communicated by Christophe Hano. Electronic supplementary material  The online version of this article (https​://doi.org/10.1007/s1124​0-020-01942​-y) contains supplementary material, which is available to authorized users. * Roland Ulber [email protected]‑kl.de 1



Institute of Bioprocess Engineering, University of Kaiserslautern, Gottlieb‑Daimler‑Str. 49‑502, 67663 Kaiserslautern, Germany

pharmacology (Sestili et al. 2018; Zhan et al. 2020). Among the pharmacologically interesting compounds found in basil are linalool (Medeiros Venancio et  al. 2016), rosmarinic (Kwon et al. 2019), oleanolic (OA; Qamar et al. 2020) and ursolic acid (UA; Arshad Qamar et al. 2010; Kümmritz et al. 2014). The latter are triterpenoids, a class of isoprenoid-ba