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

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Abstract

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.

Keymessage

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.

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Abbreviations

AA:

Artemisinic aldehyde

ADS:

Amorpha-4, 11-diene synthase

ALDH1:

Aldehyde dehydrogenase 1

APX:

Ascorbate peroxidase

Cor:

Coronatine

CSC:

Cell suspension culture

CYP71AV1:

Amorphadiene-12-hydroxylase

DBR2:

Artemisinic aldehyde Δ11(13) reductase

DCW:

Dry cell weight

DHAA:

Dihydroartemisinic aldehyde

EDTA:

Ethylenediaminetetracetic acid

GA3:

Gibberellic acid

GR:

Glutathione reductase

H2O2 :

Hydrogen peroxide

HPLC:

High-performance liquid chromatography

Kin:

Kinetin

LSD:

Least significant difference

MDA:

Malondialdehyde

MeJA:

Methyl jasmonate

NAA:

1-Naphthaleneacetic acid

PVP-40:

Polyvinylpyrrolidone

RED1:

Dihydroartemisinic aldehyde reductase

ROS:

Reactive oxygen species

SE:

Standard error

SM:

Secondary metabolite

SOD:

Superoxide dismutase

TBA:

Thiobarbituric acid

TCA:

Trichloroacetic acid

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Acknowledgements

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

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Authors

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.

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Correspondence to Maryam Salehi.

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The authors declare that they have no competing interests.

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Communicated by Ali R. Alan.

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Salehi, M., Karimzadeh, G. & Naghavi, M.R. Synergistic effect of coronatine and sorbitol on artemisinin production in cell suspension culture of Artemisia annua L. cv. Anamed. Plant Cell Tiss Organ Cult 137, 587–597 (2019). https://doi.org/10.1007/s11240-019-01593-8

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  • DOI: https://doi.org/10.1007/s11240-019-01593-8

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