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Planta Med 2009; 75(14): 1542-1547
DOI: 10.1055/s-0029-1185809
Biochemistry, Molecular Biology and Biotechnology
Original Paper
© Georg Thieme Verlag KG Stuttgart · New York

Isolation and Identification of Novel Genes Involved in Artemisinin Production from Flowers of Artemisia annua Using Suppression Subtractive Hybridization and Metabolite Analysis

Shuoqian Liu1 , Na Tian2 , Juan Li1 , Jianan Huang1 , 2 , Zhonghua Liu1 , 2
  • 1Natural Products Research Center, Hunan Agricultural University, Changsha, P. R. China
  • 2College of Horticulture and Hardening, Hunan Agricultural University, Changsha, P. R. China
Further Information

Publication History

received March 9, 2009 revised April 28, 2009

accepted May 17, 2009

Publication Date:
23 June 2009 (online)

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Abstract

Malaria is a global health problem that threatens 300–500 million people and kills more than one million people annually. Artemisinin is highly effective against multidrug-resistant Plasmodium falciparum and it has been widely used as part of the artemisinin-based combination therapies against malaria. To elucidate the biosynthetic pathway of artemisinin and to clone related genes in Artemisia annua, differentially expressed genes between blooming flowers and flower buds were isolated and characterized by a combined approach of suppression subtractive hybridization (SSH) and metabolite analysis. A total of 350 cDNA clones from a subtractive cDNA library were randomly picked, sequenced and analyzed and 253 high-quality sequences were obtained. BLASTX comparisons indicated that about 9.9 % of the clones encoded enzymes involved in isoprenoid (including artemisinin) biosynthesis. The expression of 4 gene transcripts involved in artemisinin biosynthesis was examined by RT‐PCR and the results confirmed the higher expression of these transcripts in blooming flowers than in flower buds. In addition, 2 putative transcript factors transparenta testa glabra 1 (TTG1) and ENHANCER OF GLABRA3 (GL3), which promote trichome initiation, were presented in the library. Finally, this study demonstrated that the increase of expression level of the putative TTG1 gene correlated with the improvement of glandular trichome density and artemisinin production in A. annua leaves. The subtractive cDNA library described in the present study provides important candidate genes for future research in order to increase the artemisinin content in A. annua.

Key words

Qinghaosu - sesquiterpenes - trichomes - SSH - jasmonic acid - Artemisia annua L. - Asteraceae

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Prof. Dr. Zhonghua Liu

Natural Products Research Center
Hunan Agricultural University

Changsha 410128

People's Republic of China

Phone: + 86 73 14 63 53 04

Fax: + 86 73 14 63 53 06

Email: larking_liu@yahoo.com

    www.thieme-connect.de/ejournals/toc/plantamedica