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Planta Med 2007; 73(10): 1133-1139
DOI: 10.1055/s-2007-981567
Physiology and In Vitro Biotechnology
Original Paper
© Georg Thieme Verlag KG Stuttgart · New York

Artemisinin and Sesquiterpene Precursors in Dead and Green Leaves of Artemisia annua L. Crops

Willemien J. M. Lommen1 , Sytze Elzinga1 , Francel W. A. Verstappen2 , Harro J. Bouwmeester2
  • 1Crop and Weed Ecology Group, Wageningen University, Wageningen, The Netherlands
  • 2Plant Research International, Wageningen University and Research Centre, Wageningen, The Netherlands
Further Information

Publication History

Received: February 22, 2007 Revised: May 11, 2007

Accepted: June 11, 2007

Publication Date:
12 July 2007 (online)

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Abstract

This paper analyses the accumulation and concentrations of the antimalarial artemisinin in green and dead leaves of Artemisia annua crops in two field experiments. Concentration differences were analysed as being determined by (a) the total production of artemisinin plus its upstream precursors dihydroartemisinic acid, dihydroartemisinic aldehyde, artemisinic aldehyde and artemisinic alcohol and (b) the conversion of precursors towards artemisinin. Concentrations of the total of artemisinin plus its precursors were higher in green leaves than in dead leaves in the younger crop stages, but were comparable at the final harvests. In every crop stage, the conversion of precursors to artemisinin was more advanced in dead leaves than in green leaves. This resulted in the molar concentrations of artemisinin being higher in dead leaves than in green leaves at the final harvests. The molar quantity of dihydroartemisinic acid, the last enzymatically produced precursor, was higher than that of artemisinin in green leaves, but only 19 - 27 % of that of artemisinin in dead leaves. Dead leaves were very important for the final artemisinin yield. They constituted on average 34 % of the total leaf dry matter and 47 % of the total artemisinin yield at the final harvests. The possibility to convert a larger part of dihydroartemisinic acid into artemisinin during post-harvest handling is discussed.

Key words

Artemisia annua - Asteraceae - artemisinin biosynthesis - dihydroartemisinic acid

References

  • 1 Ferreira J FS, Simon J E, Janick J. Artemisia annua: botany, horticulture, pharmacology.  Hortic Rev. 1997;  19 319-71.
    PubMedSearch in Google Scholar
  • 2 Dhingra V, Rao K V, Narasu M L. Current status of artemisinin and its derivatives as antimalarial drugs.  Life Sci. 2000;  66 279-300.
    PubMedSearch in Google Scholar
  • 3 Ferreira J FS, Simon J E, Janick J. Developmental studies of Artemisia annua: Flowering and artemisinin production under greenhouse and field conditions.  Planta Med. 1995;  61 167-70.
    Thieme ConnectPubMedSearch in Google Scholar
  • 4 Duke S O, Paul R N. Development and fine structure of the glandular trichomes of Artemisia annua L.  Int J Plant Sci. 1993;  154 107-18.
    PubMedSearch in Google Scholar
  • 5 Duke M V, Paul R N, Elsohly H N, Sturtz G, Duke S O. Localization of artemisinin and artemisitene in foliar tissues of glanded and glandless biotypes of Artemisia annua L.  Int J Plant Sci. 1994;  155 365-72.
    PubMedSearch in Google Scholar
  • 6 Bertea C M, Freije J R, van der Woude H, Verstappen F WA, Perk L, Marquez V. et al . Identification of intermediates and enzymes involved in the early steps of artemisinin biosynthesis in Artemisia annua L.  Planta Med. 2005;  71 40-7.
    Thieme ConnectPubMedSearch in Google Scholar
  • 7 Bouwmeester H J, Wallaart T E, Janssen M HA, van Loo B, Jansen B JM, Posthumus M A. et al . Amorpha-4,11-diene synthase catalyses the first probable step in artemisinin biosynthesis.  Phytochemistry. 1999;  52 843-54.
    CrossrefPubMedSearch in Google Scholar
  • 8 Teoh K H, Polichuk D R, Reed D W, Nowak G, Covello P S. Artemisia annua L. (Asteraceae) trichome-specific cDNAs reveal CYP71AV1, a cytochrome P450 with a key role in the biosynthesis of the antimalarial sesquiterpene lactone artemisinin.  FEBS Lett. 2006;  580 1411-6.
    CrossrefPubMedSearch in Google Scholar
  • 9 Ro D K, Paradise E M, Ouellet M, Fisher K J, Newman K L, Ndungu J M. et al . Production of the antimalarial drug precursor artemisinic acid in engineered yeast.  Nature. 2006;  440 940-3.
    CrossrefPubMedSearch in Google Scholar
  • 10 Wallaart T E, Pras N, Quax W J. Isolation and identification of dihydroartemisinic acid hydroperoxide from Artemisia annua: a novel biosynthetic precursor of artemisinin.  J Nat Prod. 1999;  62 1160-2.
    CrossrefPubMedSearch in Google Scholar
  • 11 Sy L -K, Brown G D. The mechanism of the spontaneous autoxidation of dihydroartemisinic acid.  Tetrahedron. 2002;  58 897-908.
    CrossrefPubMedSearch in Google Scholar
  • 12 Haynes R K. From artemisinin to new artemisinin antimalarials: biosynthesis, extraction, old and new derivatives, stereochemistry and medicinal chemistry requirements.  Curr Top Med Chem. 2006;  6 509-37.
    CrossrefPubMedSearch in Google Scholar
  • 13 Lommen W JM, Schenk E, Bouwmeester H J, Verstappen F WA. Trichome dynamics and artemisinin accumulation during development and senescence of Artemisia annua leaves.  Planta Med. 2006;  72 336-45.
    Thieme ConnectPubMedSearch in Google Scholar
  • 14 Wallaart T E, Pras N, Beekman A C, Quax W J. Seasonal variation of artemisinin and its biosynthetic precursors in plants of Artemisia annua of different geographical origin: Proof for the existence of chemotypes.  Planta Med. 2000;  66 57-62.
    Thieme ConnectPubMedSearch in Google Scholar
  • 15 Wallaart T E, Pras N, Quax W J. Seasonal variations of artemisinin and its biosynthetic precursors in tetraploid Artemisia annua plants compared with the diploid wild-type.  Planta Med. 1999;  65 723-8.
    Thieme ConnectPubMedSearch in Google Scholar
  • 16 Liersch R, Soicke H, Stehr C, Tüllner H -U. Formation of artemisinin in Artemisia annua during one vegetation period.  Planta Med. 1986;  52 387-90.
    Thieme ConnectPubMedSearch in Google Scholar
  • 17 Laughlin J C. Effect of agronomic practices on plant yield and anti-malarial constituents of Artemisia annua L.  Acta Hortic. 1993;  331 53-61.
    PubMedSearch in Google Scholar
  • 18 Morales M M, Charles D J, Simon J E. Seasonal accumulation of artemisinin in Artemisia annua L.  Acta Hortic. 1993;  344 416-20.
    PubMedSearch in Google Scholar
  • 19 Woerdenbag H J, Pras N, Nguyen Gia C han, Bui Thi B ang, Bos R, van Uden W. et al . Artemisinin, related sesquiterpenes, and essential oil in Artemisia annua during a vegetation period in Vietnam.  Planta Med. 1994;  60 272-5.
    Thieme ConnectPubMedSearch in Google Scholar
  • 20 Gupta S K, Singh P, Bajpai P, Ram G, Singh D, Gupta M M. et al . Morphogenetic variation for artemisinin and volatile oil in Artemisia annua .  Ind Crop Prod. 2002;  16 217-24.
    CrossrefPubMedSearch in Google Scholar
  • 21 Laughlin J C. The influence of distribution of antimalarial constituents in Artemisia annua L. on time and method of harvest.  Acta Hortic. 1995;  390 67-73.
    PubMedSearch in Google Scholar
  • 22 Ferreira J FS, Janick J. Distribution of artemisinin in Artemisia annua In: Janick J, editor Progress in new crops. Arlington, VA.; ASHS Press 1996: 579-84.
  • 23 Charles D J, Simon J E, Shock C C, Feibert E BG, Smith R M. Effect of water stress and post-harvest handling on artemisinin content in the leaves of Artemisia annua L. In: Janick J, Simon JE, editors New Crops. New York; Wiley 1993: 640-3.
    Search in Google Scholar
  • 24 Simonnet X, Gaudin M, Hausammann H, Vergeres C. Le fanage au champ d’Artemisia annua L.: élever la teneur en artémisinine et abaisser les coûts de production (Field drying of Artemisia annua L.: increasing artemisinin content and lowering production costs).  Revue Suisse de Viticulture, Arboriculture et Horticulture. 2001;  33 263-8.
    PubMedSearch in Google Scholar
  • 25 Laughlin J C. Post-harvest drying treatment effects on antimalarial constituents of Artemisia annua L.  Acta Hortic. 2002;  576 315-20.
    PubMedSearch in Google Scholar
  • 26 Laughlin J C, Heazlewood G N, Beattie B M. Cultivation of Artemisia annua L. In: Wright CW, editor Artemisia. London and New York; Taylor & Francis 2002: 159-95.
    Search in Google Scholar
  • 27 Knox J P, Dodge A D. Singlet oxygen and plants.  Phytochemistry. 1985;  24 889-96.
    CrossrefPubMedSearch in Google Scholar

Dr. W. J. M. Lommen

Crop and Weed Ecology Group

Wageningen University

Haarweg 333

6709 RZ Wageningen

The Netherlands

Fax: +31-317-485-572

Email: Willemien.Lommen@wur.nl