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Promotion of artemisinin content in Artemisia annua by overexpression of multiple artemisinin biosynthetic pathway genes

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Abstract

Artemisinin, isolated from an annual herbaceous plant Artemisia annua L., is an effective antimalarial compound. However, artemisinin is accumulated in small amounts (0.01–0.1% leaf dry weight) in A. annua, resulting in constant high artemisinin price. Although metabolic engineering of partial artemisinin metabolic pathway in yeast achieved great success, artemisinin from A. annua is still the important business resource. Here, we report on the generation of transgenic plants with simultaneously overexpressing four artemisinin biosynthetic pathway genes, amorpha-4,11-diene synthase gene (ADS), amorpha-4,11-diene 12-monooxygenase gene (CYP71AV1), cytochrome P450 reductase gene (CPR), and aldehyde dehydrogenase 1 gene (ALDH1) via Agrobacterium-mediated transformation. The qRT-PCR analysis demonstrated that the introduced four genes of the transgenic lines were all highly expressed. Through high-performance liquid chromatography analysis, the artemisinin contents were increased markedly in transformants, with the highest being 3.4-fold higher compared with non-converter. These results indicate that overexpression of multiple artemisinin biosynthetic pathway genes is a promising approach to improve artemisinin yield in A. annua.

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Abbreviations

ADS:

Amorpha-4,11-diene synthase

CYP71AV1:

Cytochrome P450 monooxygenase

CPR:

Cytochrome P450 reductase

ALDH1:

Aldehyde dehydrogenase1

HPLC:

High-performance liquid chromatography

PCR:

Polymerase chain reaction

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Acknowledgements

This work was funded by China National High-Tech “863” Program, Ministry of Science and Technology of the People’s Republic of China (Grant No. 2011AA100605), Shanghai Key Discipline Cultivation and Construction Project (Horticulture), and Shanghai Jiao Tong University Agri-Engineering Program.

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Authors and Affiliations

  1. Key Laboratory of Urban Agriculture (South) Ministry of Agriculture, Plant Biotechnology Research Center, Fudan-SJTU-Nottingham Plant Biotechnology R&D Center, School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai, 200240, People’s Republic of China

    Pu Shi, Xueqing Fu, Meng Liu, Qian Shen, Weimin Jiang, Ling Li, Xiaofen Sun & Kexuan Tang

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  1. Pu Shi
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  2. Xueqing Fu
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  3. Meng Liu
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Correspondence to Kexuan Tang.

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11240_2017_1173_MOESM1_ESM.pdf

Fig. S A) The mass spectrum of artemisinin standard by UPLC-MS. B) The standard curve (log-log) of artemisinin. C) The chromatographic plot of artemisinin standard. D) The chromatographic plot of representative transgenic Line 63. [M+Na]+=305.1358 (cal. 305.1365), [2M+Na]+=587.2824 (cal. 587.2832). (PDF 163 KB)

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Shi, P., Fu, X., Liu, M. et al. Promotion of artemisinin content in Artemisia annua by overexpression of multiple artemisinin biosynthetic pathway genes. Plant Cell Tiss Organ Cult 129, 251–259 (2017). https://doi.org/10.1007/s11240-017-1173-z

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