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Trichome-Specific Expression of Amorpha-4,11-diene Synthase, a Key Enzyme of Artemisinin Biosynthesis in <i>Artemisia annua</i>L., as Reported by a Promoter-GUS Fusion

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American Journal of Plant Sciences 2011年第4期2卷 619-628页

作者： Hongzhen Wang Linda Olofsson Anneli Lundgren Peter E. Brodelius 不详

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 β-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 activity 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.

关键词： Agrobacterium Tumefaciens Amorpha-4 11-diene Synthase Artemisia annua Artemisinin Biosynthesis β-Glucuronidase Gene Regulation Promoter Activity Stable Transformation

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Screening of Different <i>Artemisia</i>spp. from Western Ghats of Maharashtra for an Anti-Malarial Compound—Artemisinin

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American Journal of Plant Sciences 2015年第9期6卷 1619-1632页

作者： Vishnu Shukla Zarna Pala Anshu Alok Neetin Desai Department of Biotechnology National Agri-Food Biotechnology Institute Mohali India Department of Biological Sciences Birla Institute of Technology and Science (BITS) VidyaVihar Campus Pilani India Department of Biotechnology and Bioinformatics Dr. D. Y. Patil University Mumbai India

Artemisinin, an endoperoxide sesquiterpene lactone has proven effective in treating drug resistant cases of malaria and cancer. Artemisia annua [sweet wormwood] is the sole source for artemisinin production in many countries. To counter the low content in leaves and costly chemical synthesis process in India, alternative ways to produce artemisinin have been sought. In current study, we collected A. pallens, A. japonica and A. nilagirica from Western Ghats of Maharashtra, India and analyzed artemisinin content. Samples were extracted from leaves and florets in various extraction conditions and analyzed using different chromatographic techniques. Thin layer chromatography (TLC) and high performance thin layer chromatography (HPTLC) analysis showed the presence of compound with endoperoxide linkage in A. nilagirica. High performance liquid chromatography (HPLC) analysis showed the detection of artemisinin in methylene dichloride florets extract of A. japonica, but the concentration was too low [1.3 mg/g dry wt.] for further analyses. Gas chromatography/mass spectrometry (GC/MS) analysis identified structurally important components in the A. nilagirica ethyl acetate extract which explores the biosynthetic pathway of artemisinin from its most important precursor amorpha-4,11-diene. This is the first report of chromatographic screening of these Indian varieties of Artemisia spp. for artemisinin content.

关键词： Artemisia pallens Artemisia nilagirica Artemisia japonica Endoperoxide Amorpha-4 11-diene

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