Quantification of Three Key Enzymes Involved in Artemisinin Biogenesis in Artemisia annua by Polyclonal Antisera-Based ELISA

To elucidate the fine-tuned temporal and spatial modulation of artemisinin production in annual wormwood (Artemisia annua), we conducted enzyme-linked immunosorbent assay-based immunoquantification of three key enzymes involved in artemisinin biosynthesis, amorpha-4,11-diene synthase (ADS), cytochro...

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Bibliographic Details
Published in:Plant molecular biology reporter 2009-03, Vol.27 (1), p.50-57
Main Authors: Zeng, Qing-Ping, Zeng, Xiao-Mei, Yin, Lu-Lu, Yang, Rui-Yi, Feng, Li-Ling, Yang, Xue-Qin
Format: Article
Language:English
Subjects:
amorpha-4,11-diene synthase
Antisera
Artemisia annua
Artemisinin
biochemical pathways
Bioinformatics
Biomedical and Life Sciences
Biosynthesis
Cytochrome
cytochrome P-450
Cytochrome P450
Cytochrome P450 monooxygenase
cytochrome P450 reductase
Enzyme-linked immunosorbent assay
Enzymes
Flowering
Growth conditions
Life Sciences
Metabolomics
NADPH-ferrihemoprotein reductase
oxygenases
Plant Breeding/Biotechnology
Plant Sciences
Plants (botany)
polyclonal antibodies
Proteomics
tissue distribution
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Summary:To elucidate the fine-tuned temporal and spatial modulation of artemisinin production in annual wormwood (Artemisia annua), we conducted enzyme-linked immunosorbent assay-based immunoquantification of three key enzymes involved in artemisinin biosynthesis, amorpha-4,11-diene synthase (ADS), cytochrome P450 monooxygenase (CYP71AV1), and cytochrome P450 reductase (CPR), in various tissues and under different growth conditions. The field-grown plants accumulate abundant ADS and CYP71AV1 but a trace amount of CPR in all tested tissues. Furthermore, ADS and CYP71AV1 accumulations in leaves are 16- and eightfold higher than in roots, and ten- and fourfold higher than in stems, respectively, demonstrating a tissue-specific expression pattern. Interestingly, the flowering field plants and cold-acclimated cultural plants produce higher levels of ADS and CYP71AV1 than non-flowering field plants or untreated cultural plants, indicating the environmental and developmental induction on ADS and CYP71AV1 genes and providing possible explanation for the observation that elevation of artemisinin level occurs after flowering.
ISSN:0735-9640
1572-9818
DOI:10.1007/s11105-008-0056-1