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Trichome-Specific Expression of Amorpha-4,11-Diene Synthase, a Key Enzyme of Artemisinin Biosynthesis in Artemisia annua L., as Reported by a Promoter-GUS Fusion
Linnaeus University, Faculty of Science and Engineering, School of Natural Sciences.
Linnaeus University, Faculty of Science and Engineering, School of Natural Sciences.
Linnaeus University, Faculty of Science and Engineering, School of Natural Sciences.
Linnaeus University, Faculty of Science and Engineering, School of Natural Sciences.ORCID iD: 0000-0001-8899-5046
2011 (English)In: American Journal of Plant Sciences, ISSN 2158-2742, E-ISSN 2158-2750, Vol. 2, no 4, p. 619-628Article in journal (Refereed) Published
Abstract [en]

Artemisia annua L. produces small amounts of the sesquiterpenoid artemisinin, which is used for treatment of malaria. A worldwide shortage of the drug has led to intense research to increase the yield of artemisinin in the plant. In order to study the regulation of expression of a key enzyme of artemisinin biosynthesis, the promoter region of the key enzyme amorpha-4,11-diene synthase (ADS) was cloned and fused with the ␣-glucuronidase (GUS) reporter gene. Transgenic plants of A. annua expressing this fusion were generated and studied. Transgenic plants expressing the GUS gene were used to establish the activity of the cloned promoter by a GUS activity staining procedure. GUS under the control of the ADS promoter showed specific expression in glandular trichomes. The activity of the ADS promoter varies temporally and in old tissues essentially no GUS staining could be observed. The expression pattern of GUS and ADS in aerial parts of the transgenic plant was essentially the same indicating that the cis-elements controlling glandular trichome specific expression are included in the cloned promoter. However, some cis-element(s) that control expression in root and old leaf appears to be missing in the cloned promoter. Furthermore, qPCR was used to compare the activity of the wild-type ADS promoter with that of the cloned ADS promoter. The latter promoter showed a considerably lower activ- ity than the wild-type promoter as judged from the levels of GUS and ADS transcripts, respectively, which may be due to the removal of an enhancing cis-element from the ADS promoter. The ADS gene is specifically expressed in stalk and secretory cells of glandular trichomes of A. annua.

Place, publisher, year, edition, pages
2011. Vol. 2, no 4, p. 619-628
Keywords [en]
Agrobacterium Tumefaciens, Amorpha-4, 11-Diene Synthase, Artemisia annua, Artemisinin Biosynthesis, ␣-Glucuronidase, Gene Regulation, Promoter Activity, Stable Transformation
National Category
Biochemistry and Molecular Biology
Research subject
Natural Science, Biochemistry
Identifiers
URN: urn:nbn:se:lnu:diva-18065DOI: 10.4236/ajps.2011.24073OAI: oai:DiVA.org:lnu-18065DiVA, id: diva2:511107
Available from: 2012-03-20 Created: 2012-03-20 Last updated: 2017-12-07Bibliographically approved

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Wang, HongzhenOlofsson, LindaLundgren, AnneliBrodelius, Peter E.

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