Synergistic effect of coronatine and sorbitol on artemisinin production in cell suspension culture of Artemisia annua L. cv. Anamed

  • Maryam SalehiEmail author
  • Ghasem Karimzadeh
  • Mohammad Reza Naghavi
Original Article


Artemisinin is an efficient anti-malarial drug and it possesses biological activity against a wide range of cancers. The combined application of two different elicitors can be an efficient way to increase the production of secondary metabolite in plant cell cultures. The results of coronatine (Cor) pretreatment and three concentrations of sorbitol were assessed on the growth, biochemical traits, expression of artemisinin biosynthetic genes, and artemisinin production in Artemisia annua cell suspension culture (CSC). After pretreating CSC with 0.05 µM Cor [on the 14th day (three days before the stationary phase) for 48 h], liquid medium in the culture flasks was decanted and replaced with fresh medium (containing 30 g/L sucrose) plus or minus sorbitol at selected concentrations (0, 20, 30, and 40 g/L) on day 16th (one day before the stationary phase). The sorbitol treatment enhanced the contents of malondialdehyde (MDA) and hydrogen peroxide (H2O2) and resulted in oxidative stress. Cor-pretreatment increased the activity of antioxidant enzymes and consequently it reduced H2O2 content and oxidative stress which resulted in decreased MDA content and better growth. The application of Cor plus sorbitol resulted in a dramatic enhancement in the expression of artemisinin biosynthetic genes and artemisinin production at all concentrations. The expression levels of artemisinin biosynthetic genes (about 7.66, 8.67, 8.67, and 8.33-fold in ADS, CYP71AV1, ALDH1, and DBR2 genes, respectively at 4 h after sorbitol treatment) and artemisinin production (9.33 mg/L, 8-fold) peaked at 30 g/L sorbitol plus Cor and decreased at 40 g/L sorbitol, probably because of higher oxidative stress.


The simultaneous application of Cor and sorbitol resulted in a dramatic enhancement in the expression of artemisinin biosynthetic genes and artemisinin production owing to a synergistic or potentiating result.


Antioxidant enzyme Osmotic stress Relative gene expression Secondary metabolite 



Artemisinic aldehyde


Amorpha-4, 11-diene synthase


Aldehyde dehydrogenase 1


Ascorbate peroxidase




Cell suspension culture




Artemisinic aldehyde Δ11(13) reductase


Dry cell weight


Dihydroartemisinic aldehyde


Ethylenediaminetetracetic acid


Gibberellic acid


Glutathione reductase


Hydrogen peroxide


High-performance liquid chromatography




Least significant difference




Methyl jasmonate


1-Naphthaleneacetic acid




Dihydroartemisinic aldehyde reductase


Reactive oxygen species


Standard error


Secondary metabolite


Superoxide dismutase


Thiobarbituric acid


Trichloroacetic acid



Authors gratefully acknowledge the support provided for this survey by the Tarbiat Modares University, Tehran, Iran.

Author Contributions

Maryam Salehi designed and performed experiments and prepared the manuscript under the joint supervision of Assoc. prof. G. Karimzadeh and Prof. M. R. Naghavi. All authors read and approved the final manuscript.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no competing interests.


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Copyright information

© Springer Nature B.V. 2019

Authors and Affiliations

  • Maryam Salehi
    • 1
    Email author
  • Ghasem Karimzadeh
    • 1
  • Mohammad Reza Naghavi
    • 2
  1. 1.Department of Plant Genetics and Breeding, Faculty of AgricultureTarbiat Modares UniversityTehranIran
  2. 2.Department of Agronomy and Plant Breeding, College of Agricultural and Natural ResourcesUniversity of TehranKarajIran

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