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Enhancement of artemisinin content and biomass in Artemisia annua by exogenous GA3 treatment

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Abstract

Artemisinin is a promising and potent antimalarial drug naturally produced by the plant Artemisia annua L. but in very low yield. Its artemisinin content is known to be greatly affected by both genotype and environmental factors. In this study, the production of artemisinin and leaf biomass in Artemisia annua L. was significantly increased by exogenous GA3 treatment. The effect of GA3 application on expression of proposed key enzymes involved in artemisinin yield was examined in both wild type (007) and FPS-overexpression (253-2) lines of A. annua. In the wild type (007) at 6 h post GA3 application there was an abrupt rise in FPS, ADS and CYP71AV1 expression and at 24 h a temporary and significant peak in artemisinin (1.45-fold higher than the control). After GA3 application in line 253-2, there was a dramatic rise in expression of FPS at 3 h, CYP71AV1 at 9 h and ADS at 72 h and accumulation of artemisinin after 7 days, which was a delay when compared with the wild type plant. Thus, increased artemisinin content from exogenous GA3 treatment was associated with increased expression of key enzymes in the artemisinin biosynthesis pathway. Interestingly, exogenous GA3 continuously enhanced artemisinin content from the vegetative stage to flower initiation in both plant lines and gave significantly higher leaf biomass than in control plants. Consequently, the artemisinin yield in GA3-treated plants was much higher than in control plants. Although the maximum artemisinin content was found at the full blooming stage [2.1% dry weight (DW) in 007 and 2.4% DW in 253-2], the highest artemisinin yield in GA3-treated plants was obtained during the flower initiation stage (2.4 mg/plant in 007 and 2.3 mg/plant in 235-2). This was 26.3 and 27.8% higher, respectively, than in non-treated plants 007 and 253-2. This study showed that exogenous GA3 treatment enhanced artemisinin production in pot experiments and should be suitable for field application.

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Abbreviations

ADS:

Amorpha-4,11-diene synthase

CYP71AV1:

Cytochrome P450 monooxygenase

DW:

Dry weight

EF1-α:

Elongation factor 1 alpha

FPS:

Farnesyl pyrophosphate synthase

FW:

Fresh weight

GA:

Gibberellic acid

qPCR:

Quantitative Real-time PCR

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Acknowledgments

This research was supported by Mahidol University grant, Thailand. The authors would like to thank the scholarship from The Royal Golden Jubilee PhD Program (RGJ) of the Thailand Research Fund. This work was conducted under the collaboration of Mahidol University and Chiba University for double degree program. Thanks to Prof. Timothy W. Flegel and FGS, MU for English language proof of the manuscript.

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Authors and Affiliations

  1. Department of Biotechnology, Faculty of Science, Mahidol University, Bangkok, 10400, Thailand

    Waleerat Banyai & Kanyaratt Supaibulwatana

  2. Laboratory of Plant Cell Technology, Faculty of Horticulture, Chiba University, Chiba, 271-8510, Japan

    Waleerat Banyai & Masahiro Mii

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  1. Waleerat Banyai
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  2. Masahiro Mii
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  3. Kanyaratt Supaibulwatana
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Correspondence to Kanyaratt Supaibulwatana.

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Banyai, W., Mii, M. & Supaibulwatana, K. Enhancement of artemisinin content and biomass in Artemisia annua by exogenous GA3 treatment. Plant Growth Regul 63, 45–54 (2011). https://doi.org/10.1007/s10725-010-9510-9

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