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Characterization of development and artemisinin biosynthesis in self-pollinated Artemisia annua plants

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Abstract

Artemisia annua L. is the only natural resource that produces artemisinin (Qinghaosu), an endoperoxide sesquiterpene lactone used in the artemisinin-combination therapy of malaria. The cross-hybridization properties of A. annua do not favor studying artemisinin biosynthesis. To overcome this problem, in this study, we report on selection of self-pollinated A. annua plants and characterize their development and artemisinin biosynthesis. Self-pollinated F2 plants selected were grown under optimized growth conditions, consisting of long day (16 h of light) and short day (9 h of light) exposures in a phytotron. The life cycles of these plants were approximately 3 months long, and final heights of 30–35 cm were achieved. The leaves on the main stems exhibited obvious morphological changes, from indented single leaves to odd, pinnately compound leaves. Leaves and flowers formed glandular and T-shaped trichomes on their surfaces. The glandular trichome densities increased from the bottom to the top leaves. High performance liquid chromatography–mass spectrometry-based metabolic profiling analyses showed that leaves, flowers, and young seedlings of F2 plants produced artemisinin. In leaves, the levels of artemisinin increased from the bottom to the top of the plants, showing a positive correlation to the density increase of glandular trichomes. RT-PCR analysis showed that progeny of self-pollinated plants expressed the amorpha-4, 11-diene synthase (ADS) and cytochrome P450 monooxygenase 71 AV1 (CYP71AV1) genes, which are involved in artemisinin biosynthesis in leaves and flowers. The use of self-pollinated A. annua plants will be a valuable approach to the study of artemisinin biosynthesis.

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Abbreviations

ADS:

Amorpha-4, 11-diene

CYP71AV1:

Cytochrome P450 monooxygenase 71 AV1

HPLC–ESI–MS:

High performance liquid chromatography–electron spray ionization–mass spectrometry

MS:

Murashige and Skoog

QHS:

Qinghaosu

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Acknowledgments

We thank the North Carolina Biotechnology Center for providing this research grant (Grant #55031) to support the genetic engineering of the artemisinin research. We give our special gratitude to Dr. George Yatskievych from Missouri Herbarium at Missouri Botanical Garden for his kind help to collect Artemisia annua seeds. We thank Dr. Valerie Knowlton from the Center for Electron Microscopy for her kind help in preparing the SEM.

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  1. Department of Plant Biology, North Carolina State University, Raleigh, NC, 27695, USA

    Fatima Alejos-Gonzalez, Guosheng Qu, Li-Li Zhou, Carole H. Saravitz, Janet L. Shurtleff & De-Yu Xie

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  1. Fatima Alejos-Gonzalez
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  2. Guosheng Qu
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  3. Li-Li Zhou
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Correspondence to De-Yu Xie.

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Alejos-Gonzalez, F., Qu, G., Zhou, LL. et al. Characterization of development and artemisinin biosynthesis in self-pollinated Artemisia annua plants. Planta 234, 685–697 (2011). https://doi.org/10.1007/s00425-011-1430-z

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