Plant Cell, Tissue and Organ Culture (PCTOC)

, Volume 103, Issue 2, pp 255–265

Overexpression of farnesyl pyrophosphate synthase (FPS) gene affected artemisinin content and growth of Artemisia annua L.

  • Waleerat Banyai
  • Chalermpol Kirdmanee
  • Masahiro Mii
  • Kanyaratt Supaibulwatana
Original Paper

DOI: 10.1007/s11240-010-9775-8

Cite this article as:
Banyai, W., Kirdmanee, C., Mii, M. et al. Plant Cell Tiss Organ Cult (2010) 103: 255. doi:10.1007/s11240-010-9775-8

Abstract

Transgenic plants of Artemisia annua L., a medicinal plant that produces the compound artemisinin which has an anti-malarial activity, were developed following Agrobacterium tumefaciens-mediated transformation of leaf explants. A. tumefaciens strain EHA105 carrying either pCAMBIA1301 or pCAMBIAFPS was used. Both plasmids harbored the hygromycin phosphotransferase II (hptII) gene as a selectable gene, but the latter plasmid also harbored the gene encoding for farnesyl pyrophosphate synthase (FPS), a key enzyme for artemisinin biosynthesis. Shoot regeneration was observed either directly from leaf sections or via intervening callus when explants were incubated on solidified Murashige and Skoog (MS) (1962) medium containing 0.1 mg l−1 α-naphthaleneacetic acid (NAA), 1 mg l−1 N6-benzyladenine (BA), 30 mg l−1 meropenem and 10 mg l−1 hygromycin. Applying vacuum infiltration dramatically increased transformation efficiency up to 7.3 and 19.7% when plasmids with and without FPS gene were used, respectively. All putative transgenic regenerants showed positive bands of hptII gene following Southern blot analysis. Expression of FPS was observed in all transgenic lines, and FPS over-expressed lines exhibited higher artemisinin content and yield, of 2.5- and 3.6-fold, respectively, than that detected in wild-type plants. A relatively high correlation (R2 = 0.78) was observed between level of expression of FPS and artemisinin content. However, gene silencing was detected in some transgenic lines, especially for those lines containing two copies of the FPS transgene, and with some lines exhibiting reduced growth.

Keywords

Artemisia annua L. Vacuum infiltration Agrobacterium tumefaciens Transformation Farnesyl pyrophosphate synthase 

Abbreviations

FPS

Farnesyl pyrophosphate synthase gene

GUS

β-glucuronidase

hptII

Hygromycin phosphotransferase gene

NAA

α-naphthaleneacetic acid

BA

N6-benzyladenine

PGR

Plant growth regulators

FW

Fresh weight

DW

Dry weight

Copyright information

© Springer Science+Business Media B.V. 2010

Authors and Affiliations

  • Waleerat Banyai
    • 1
    • 3
  • Chalermpol Kirdmanee
    • 2
  • Masahiro Mii
    • 3
  • Kanyaratt Supaibulwatana
    • 1
  1. 1.Department of Biotechnology, Faculty of ScienceMahidol UniversityBangkokThailand
  2. 2.National Center For Genetic Engineering and BiotechnologyNational Science and Technology Development AgencyPathumthaniThailand
  3. 3.Laboratory of Plant Cell Technology, Faculty of HorticultureChiba UniversityChibaJapan

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