Staff Publications

Staff Publications

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    'Staff publications' is the digital repository of Wageningen University & Research

    'Staff publications' contains references to publications authored by Wageningen University staff from 1976 onward.

    Publications authored by the staff of the Research Institutes are available from 1995 onwards.

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Record number 448216
Title The metabolite chemotype of Nicotiana benthamiana transiently expressing artemisinin biosynthetic pathway genes is a function of CYP71AV1 type and relative gene dosage
Author(s) Ting, H.M.; Wang, B.; Ryden, A.M.; Woittiez, L.S.; Verstappen, F.W.A.; Ruyter-Spira, C.P.; Beekwilder, M.J.; Bouwmeester, H.J.; Krol, A.R. van der; Herpen, T.W.J.M. van
Source New Phytologist 199 (2013)2. - ISSN 0028-646X - p. 352 - 366.
DOI https://doi.org/10.1111/nph.12274
Department(s) Laboratory of Plant Physiology
Plant Production Systems
BIOS Applied Metabolic Systems
EPS
Publication type Refereed Article in a scientific journal
Publication year 2013
Keyword(s) molecular-cloning - annua l - amorpha-4,11-diene synthase - dihydroartemisinic acid - mass-spectrometry - key role - arabidopsis - identification - metabolomics - precursors
Abstract Artemisia annua, which produces the anti-malaria compound artemisinin, occurs as high-artemisinin production (HAP) and low-artemisinin production (LAP) chemotypes. Understanding the basis of the difference between these chemotypes would assist breeding and optimising artemisinin biosynthesis. Here we present a systematic comparison of artemisinin biosynthesis genes that may be involved in determining the chemotype (CYP71AV1, DBR2 and ALDH1). These genes were isolated from the two chemotypes and characterized using transient expression in planta. The enzyme activity of DBR2 and ALDH1 from the two chemotypes did not differ, but structural differences in CYP71AV1 from LAP and HAP chemotypes (AMOLAP and AMOHAP, respectively) resulted in altered enzyme activity. AMOLAP displays a seven amino acids N-terminal extension compared with AMOHAP. The GFP fusion of both proteins show equal localization to the ER but AMOHAP may have reduced stability. Upon transient expression in Nicotiana benthamiana, AMOLAP displayed a higher enzyme activity than AMOHAP. However, expression in combination with the other pathway genes also resulted in a qualitatively different product profile ('chemotype'); that is, in a shift in the ratio between the unsaturated and saturated (dihydro) branch of the pathway.
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