title: Expression of β-glucosidase increases trichome density and artemisinin content in transgenic Artemisia annua plants.
abstract: Artemisinin is highly effective against multidrug-resistant strains of Plasmodium falciparum, the aetiological agent of the most severe form of malaria. However, a low level of accumulation of artemisinin in Artemisia annua is a major limitation for its production and delivery to malaria endemic areas of the world. While several strategies to enhance artemisinin have been extensively explored, enhancing storage capacity in trichome has not yet been considered. Therefore, trichome density was increased with the expression of β-glucosidase (bgl1) gene in A. annua through Agrobacterium-mediated transformation. Transgene (bgl1) integration and transcript were confirmed by molecular analysis. Trichome density increased up to 20% in leaves and 66% in flowers of BGL1 transgenic plants than Artemisia control plants. High-performance liquid chromatography, time of flight mass spectrometer data showed that artemisinin content increased up to 1.4% in leaf and 2.56% in flowers (per g DW), similar to the highest yields achieved so far through metabolic engineering. Artemisinin was enhanced up to five-fold in BGL1 transgenic flowers. This study opens the possibility of increasing artemisinin content by manipulating trichomes' density, which is a major reservoir of artemisinin. Combining biosynthetic pathway engineering with enhancing trichome density may further increase artemisinin yield in A. annua. Because oral feeding of Artemisia plant cells reduced parasitemia more efficiently than the purified drug, reduced drug resistance and cost of prohibitively expensive purification process, enhanced expression should play a key role in making this valuable drug affordable to treat malaria in a large global population that disproportionally impacts low-socioeconomic areas and underprivileged children.
authors.affiliation: Department of Biochemistry, School of Dental Medicine, University of Pennsylvania, Philadelphia, PA, USA.
authors.forename: Nameirakpam Dolendro
authors.initials: ND
authors.lastname: Singh
authors.affiliation: Department of Biochemistry, School of Dental Medicine, University of Pennsylvania, Philadelphia, PA, USA.
authors.forename: Shashi
authors.initials: S
authors.lastname: Kumar
authors.affiliation: Department of Biochemistry, School of Dental Medicine, University of Pennsylvania, Philadelphia, PA, USA.
authors.forename: Henry
authors.initials: H
authors.lastname: Daniell
mesh_terms: D031172:Artemisia annua / Q000201:enzymology* / Q000235:genetics* / Q000648:ultrastructure; D037621:Artemisinins / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid; D018628:Gene Dosage; D018515:Plant Leaves / Q000648:ultrastructure; D030821:Plants, Genetically Modified; D014170:Transformation, Genetic; D019076:Transgenes; D064201:Trichomes / Q000254:growth & development* / Q000648:ultrastructure; D001617:beta-Glucosidase / Q000378:metabolism*
chemical_list: D037621:Artemisinins; C031327:artemisinin; D001617:beta-Glucosidase
keywords: artemisinin storage; genetic modification; global health; infectious disease; malaria