The metabolite chemotype of Nicotiana benthamiana transiently expressing artemisinin biosynthetic pathway genes is a function of CYP71AV1 type and relative gene dosage
Journal
New Phytologist
Journal Volume
199
Journal Issue
2
Pages
352
Date Issued
2013-07-01
Author(s)
Wang, Bo
Rydén, Anna Margareta
Woittiez, Lotte
Van Herpen, Teun
Verstappen, Francel W A
Ruyter-Spira, Carolien
Beekwilder, Jules
Bouwmeester, Harro J.
Van der Krol, Alexander
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. © 2013 The Authors New Phytologist © 2013 New Phytologist Trust.
Subjects
Artemisia annua | Artemisinin | CYP71AV1 | Nicotiana benthamiana | Transient expression
SDGs
Type
journal article
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