Abstract
Artemisinin is widely used as an antimalarial drug, and the regulation of artemisinin metabolism is at the forefront of artemisinin research. A WRKY transcription factor, named as AaWRKY4, was cloned from high artemisinin-yielding Artemisia annua, which has similar expression pattern with the key enzymes in artemisinin biosynthetic pathway. AaWRKY4 was preferentially expressed in glandular secretory trichomes (GSTs) of young leaves and flower buds, but weakly expressed in other tissues. To further study the function of AaWRKY4, plant expression vector pHB-AaWRKY4 containing AaWRKY4 driven by CaMV 35S promoter was constructed and introduced into A. annua via Agrobacterium tumafeciens-mediated transformation. Expression analysis showed that the expression of AaWRKY4 was increased in transgenic plants. Four independent transgenic plants overexpressing AaWRKY4 were selected for further analysis. The expression levels of artemisinin biosynthetic pathway genes ADS, CYP71AV1, DBR2 and ALDH1 were dramatically increased in AaWRKY4-overexpressing A. annua plants. Furthermore, the artemisinin yield was increased by 35–50% in AaWRKY4-overexpressing A. annua plants. These results indicate AaWRKY4 can upregulate artemisinin content through regulating artemisinin metabolism.
Key message
A WRKY transcription factor, named as AaWRKY4, was cloned from high artemisinin-yielding Artemisia annua. AaWRKY4 was preferentially expressed in glandular secretory trichomes (GSTs) of young leaves and flower buds. The expression levels of artemisinin biosynthetic pathway genes ADS, CYP71AV1, DBR2 and ALDH1 were dramatically increased in AaWRKY4-overexpressing A. annua plants. Furthermore, the artemisinin yield was increased by 35–50% in AaWRKY4- overexpressing A. annua plants.
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Data availability
The Gene Bank number for AaWRKY4 is MW169031. Sequence data from this article can be found in the Arabidopsis Genome Initiative or the GenBank databases.
Abbreviations
- GSTs:
-
Glandular secretory trichomes
- DMAPP:
-
Dimethylallyl diphosphate
- IPP:
-
Isopentenyl diphosphate
- MVA pathway:
-
Mevalonate pathway
- MEP pathway:
-
Nonmevalonate pathway
- ADS:
-
Amorpha-4, 11-diene synthase
- CYP:
-
Cytochrome P450 enzyme CYP71AV1
- DBR2:
-
Artemisinic aldehyde ∆11 (13) reductase
- ALDH1:
-
Aldehyde dehydrogenase
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Acknowledgements
This work was supported by Hunan Provincial Natural Science Foundation of China (Grant No. 2018JJ3008), NSF of Anhui Province (Grant No. 1908085MH268), SJTU Trans-med Awards Research (Grant No. 20190104), the Open Fund Project of Key Laboratory in Hunan Universities (Grant No. NY20K04), Science Foundation of Hengyang Normal University (Grant No. 17D14), and Science Foundation of Hengyang Normal University (Grant No. 17A03).
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WJ and KT conceived and designed the project. HH and XH conducted the experiments and analyzed the data. HH and WJ drafted the paper. WJ and KT reviewed the manuscript. All authors read and approved the final manuscript.
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Huang, H., Xing, S., Tang, K. et al. AaWRKY4 upregulates artemisinin content through boosting the expressions of key enzymes in artemisinin biosynthetic pathway. Plant Cell Tiss Organ Cult 146, 97–105 (2021). https://doi.org/10.1007/s11240-021-02049-8
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DOI: https://doi.org/10.1007/s11240-021-02049-8