Abstract
Rosmarinic acid (RA), as the major secondary metabolite of Salvia species, is a valuable bioactive compound for pharmaceutical applications. Toward rosmarinic acid production, Murashige and Skoog (MS) medium supplemented with 0.5 mg L−1 2,4-D and 2 mg L−1 BA or 1 mg L−1 2,4-D and 1 mg L−1 BA showed the highest percentage of callus induction (100%). In this research, MS medium supplemented with various concentrations of sucrose, phenylalanine, salicylic acid (SA), and methyl jasmonate (MeJA) were evaluated using cell suspension cultures of Salvia nemorosa L. Sucrose treatment (4.5%) increased cell biomass (20.58 g L−1) and RA content (17.1 mg g−1 DW). Phenylalanine had no significant effect on cell biomass; however, phenylalanine led to a higher accumulation of RA (16.2 mg g−1 DW) compared to non-treated cells. MeJA caused a slight decrease in cell growth. Maximum RA content (14.2 mg g−1 DW) observed in cells treated with 200 μM MeJA, 48 h after elicitation. SA (0.5 μM) treatment showed an impressive effect on RA production (26.4 mg g−1 DW) with an increase of 8 times compared to the control. Phenylalanine ammonia-lyase and RA synthase expression were positively correlated with RA biosynthesis after induction with SA. Results showed that SA might be considered as an economical elicitor compound for large-scale production of RA.
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The authors would like to acknowledge the University of Tehran Vice Chancellor and the College of Agriculture & Natural Resources Vice Chancellor for supporting this research. The authors wish to thank Mr. Hamid Ahadi for his kind help in HPLC analysis.
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Fatemeh Khoshsokhan, Mesbah Babalar, and Mohammad Hossein Mirjalili conceived and designed the experiments. Material preparation, data collection, and analysis were performed by Fatemeh Khoshsokhan, Mesbah Babalar, Seyed Alireza Salami, and Reza Sheikhakbari-Mehr. The first draft of the manuscript was written by Fatemeh Khoshsokhan, and all authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.
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Khoshsokhan, F., Babalar, M., Salami, S.A. et al. An efficient protocol for production of rosmarinic acid in Salvia nemorosa L.. In Vitro Cell.Dev.Biol.-Plant 59, 298–314 (2023). https://doi.org/10.1007/s11627-023-10328-6
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DOI: https://doi.org/10.1007/s11627-023-10328-6