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Cited by 47Plant biochemistry and biosynthesisPlant tissue culture and regenerationSesquiterpenes and Asteraceae Studies

Enhanced artemisinin yield by expression of rol genes in Artemisia annua.

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Dilshad, Erum, Cusido, Rosa Maria, Palazon, Javier, Estrada, Karla Ramirez, Bonfill, Mercedes, Mirza, Bushra·2015·Malaria Journal
DOI 10.1186/s12936-015-0951-5PMC PMC4625606
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Abstract

BACKGROUND: Despite of many advances in the treatment of malaria, it is still the fifth most prevalent disease worldwide and is one of the major causes of death in the developing countries which accounted for 584,000 deaths in 2013, as estimated by World Health Organization. Artemisinin from Artemisia annua is still one of the most effective treatments for malaria. Increasing the artemisinin content of A. annua plants by genetic engineering would improve the availability of this much-needed drug. METHODS: In this regard, a high artemisinin-yielding hybrid of A. annua produced by the centre for novel agricultural products of the University of York, UK, was selected (artemisinin maximally 1.4 %). As rol genes are potential candidates of biochemical engineering, genetic transformation of A. annua with Agrobacterium tumefaciens GV3101 harbouring vectors with rol B and rol C genes was carried out with the objective of enhancement of artemisinin content. Transgenic lines produced were analysed by the LC-MS for quantitative analysis of artemisinin and analogues. These high artemisinin yielding transgenics were also analysed by real time quantitative PCR to find the molecular dynamics of artemisinin enhancement. Genes of artemisinin biosynthetic pathway were studied including amorphadiene synthase (ADS), cytochrome P450, (CYP71AV1) and aldehyde dehydrogenase 1 (ALDH1). Trichome-specific fatty acyl-CoA reductase 1(TAFR1) is an enzyme involved in both trichome development and sesquiterpenoid biosynthesis and both processes are important for artemisinin biosynthesis. Thus, real time qPCR analysis of the TAFR1 gene was carried out, and trichome density was determined. RESULTS: Transgenics of rol B gene showed two- to ninefold (the decimal adds nothing in the abstract, please simplify to two- to ninefold) increase in artemisinin, 4-12-fold increase in artesunate and 1.2-3-fold increase in dihydroartemisinin. Whereas in the case of rol C gene transformants, a fourfold increase in artemisinin, four to ninefold increase in artesunate and one- to twofold increase in dihydroartemisinin concentration was observed. Transformants with the rol B gene had higher expression of these genes than rol C transformants. TAFR1 was also found to be more expressed in rol gene transgenics than wild type A. annua, which was also in accordance with the trichome density of the respective plant. CONCLUSION: Thus it was proved that rol B and rol C genes are effective in the enhancement of artemisinin content of A. annua, rol B gene being more active to play part in this enhancement than rol C gene.

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  • Transformation of A. annua with rolB and rolC genes of Agrobacterium rhizogenes led to plants with increased content of artemisinin.

    “It has been recently shown that the transformation of A. annua with the rolB and rolC genes of Agrobacterium rhizogenes led to plants with an increased content of artemisinin (Dilshad et al., 2015).”

    From Cannabis sativa: The Plant of the Thousand and One Molecules (2016) · cites this paper as Dilshad et al., 2015

  • In A. annua, rolB and rolC trigger different effects, with rolB showing enhanced production with respect to rolC.

    “The study on A. annua showed that rolB and rolC trigger different effects, with rolB showing enhanced production with respect to rolC.”

    From Cannabis sativa: The Plant of the Thousand and One Molecules (2016) · cites this paper as Dilshad et al., 2015

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Cited by

Sorted by citations

Authors
Dilshad, E., Cusido, R. M., Palazon, J., Estrada, K. R., Bonfill, M., & Mirza, B.
Affiliations
  • Quaid-i-Azam University (PK)
  • Universitat de Barcelona (ES)
Corresponding author
Mirza, Bushra
ORCID
Venue
Malaria Journal
Publisher
Springer Science and Business Media LLC
Published
2015-10-29
Volume
14
Issue
1
Pages
424-424
ISSN
1475-2875 (ISSN-L)
DOI
10.1186/s12936-015-0951-5
PMC
PMC4625606
OpenAlex
W1963072412
Cited by
47
References
6
Citations by year
2026: 2 · 2025: 1 · 2024: 2 · 2023: 9 · 2022: 5 · 2021: 7 · 2020: 3 · 2019: 1 · 2018: 6 · 2017: 3 · 2016: 8
FWCI
2.46
Type
journal article
Language
en
Open access
Licence
cc-by
Locations
  • Malaria Journal (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • RECERCAT (Consorci de Serveis Universitaris de Catalunya) (publishedVersion, other-oa)
  • Dipòsit Digital de la Universitat de Barcelona (Universitat de Barcelona) (submittedVersion, cc-by)
Funders
  • Generalitat de Catalunya (2014SGR215, BIO2014-51861-R)
  • Ministerio de Economía y Competitividad (BIO2014-51861-R)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Plant biochemistry and biosynthesis
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Plant tissue culture and regeneration
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cancer Research › Sesquiterpenes and Asteraceae Studies
Keywords
artemisinin, Artemisia annua, rol genes, rolB, rolC, transgenic plants, artemisinin biosynthesis, trichome development, sesquiterpene biosynthesis, CYP71AV1, ALDH1, artesunate, dihydroartemisinin
MeSH terms
Antimalarials, Chromatography, Liquid, Mass Spectrometry, Transformation, Genetic, Gene Expression, Agrobacterium tumefaciens, Genes, Plant, Plants, Genetically Modified, Artemisia annua, Artemisinins, Metabolic Networks and Pathways, Metabolic Engineering, Real-Time Polymerase Chain Reaction

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