Establishment of recombinant Catharanthus roseus stem cells stably overexpressing ORCA4 for terpenoid indole alkaloids biosynthesis

• Published in: Plant physiology and biochemistry 2023-03, Vol.196, p.783-792
• Main Authors: Yang, Yuanjian, Ding, Liuyu, Zhou, Ying, Guo, Zizheng, Yu, Rongmin, Zhu, Jianhua
• Format: Article
• Language: English
• Subjects: Cambial meristematic cells; Catharanthus - genetics; Catharanthus roseus; Indole Alkaloids - metabolism; Indole Alkaloids - pharmacology; Octadecanoid-derivative responsive Catharanthus AP2-domain protein 3; Octadecanoid-derivative responsive Catharanthus AP2-domain protein 4; Secologanin Tryptamine Alkaloids - metabolism; Secologanin Tryptamine Alkaloids - pharmacology; Terpenoid indole alkaloids; Transcription Factors - metabolism; Vinblastine - metabolism; Vinblastine - pharmacology; Vincristine - metabolism; Vincristine - pharmacology
• ISSN: 0981-9428; 1873-2690
• DOI: 10.1016/j.plaphy.2023.02.039

Catharanthus roseus is a perennial herb of the Apocynaceae family, from which about 200 kinds of alkaloids have been characterized. Most alkaloids from C. roseus are terpenoid indole alkaloids (TIAs), such as vinblastine and vincristine, which are widely used in the clinic for their good antitumor activity. However, they were only biosynthesized in C. roseus, and their content in C. roseus is extremely low. The access to these valuable compounds is by plant extraction or chemical semisynthesis from their precursors catharanthine and vindoline. Since catharanthine and vindoline are also obtained from C. roseus, the supply of vinblastine and vincristine makes it difficult to meet market demands. Therefore, how to improve the yield of TIAs is an attractive issue. In this study, we compared the regulatory effect of two critical transcription factors, octadecanoid-derivative responsive Catharanthus AP2-domain protein 3 (ORCA3) and octadecanoid-derivative responsive Catharanthus AP2-domain protein 4 (ORCA4), on the biosynthesis of TIAs in C. roseus. The results showed that overexpressing both two transcription factors could increase the accumulation of TIAs. The effect was more significant when ORCA4 was overexpressed. To acquire C. roseus TIAs on a continuous and consistent basis, we then created and acquired C. roseus stem cells stably overexpressing ORCA4. This is the first time a recombinant C. roseus stem cell system with stable ORCA4 overexpression has been developed, which not only provides new ideas for future research in this area but also breaches new life into the industrial application of using plant cell culture to obtain natural products. [Display omitted] •Transcription factor ORCA3 regulated the expression of ORCA4 in Catharanthus roseus.•ORCA4 overexpression resulted in accumulation of catharanthine and vindoline.•C. roseus stem cells overexpressing ORCA4 were built for the first time.•Overexpressing ORCA4 is beneficial for terpenoid indole alkaloids biosynthesis.