Short Communication A New Geranylgeranyl Diphosphate Synthase Gene from Ginkgo biloba, which Intermediates the Biosynthesis of the Key Precursor for Ginkgolides ZHIHUA LIAO a , MIN CHEN b , YIFU GONG c,d , LIANG GUO a , QIUMIN TAN a , XIAOQI FENG a , XIAOFEN SUN a , FENG TAN e and KEXUAN TANG a,c, * a State Key Laboratory of Genetic Engineering, Institute of Genetics, School of Life Sciences, Morgan-Tan International Center for Life Sciences, Fudan- SJTU-Nottingham Plant Biotechnology R&D Center, Fudan University, Shanghai 200433, People’s Republic of China; b School of Pharmacy, Fudan University, Shanghai 200032, People’s Republic of China; c Plant Biotechnology Research Center, Fudan-SJTU-Nottingham Plant Biotechnology R&D Center, School of Agriculture and Biology, Shanghai Jiaotong University, Shanghai 200030, People’s Republic of China; d Faculty of Life Science and Biotechnology, Ningbo University, Ningbo 315211, People’s Republic of China; e School of Life Sciences, Southwest China Normal University, Chongqing 400715, People’s Republic of China (Received 6 October 2003) Geranylgeranyl diphosphate synthase (GGPPS, EC: 2.5.1.29) catalyzes the biosynthesis of geranylgeranyl diphosphate (GGPP), which is a key precursor for ginkgolide biosynthesis. Here we reported for the first time the cloning of a new full-length cDNA encoding GGPPS from the living fossil plant Ginkgo biloba. The full-length cDNA encoding G. biloba GGPPS (designated as GbGGPPS) was 1657 bp long and contained a 1176 bp open reading frame encoding a 391 amino acid protein. Comparative analysis showed that GbGGPPS possessed a 79 amino acid transit peptide at its N-terminal, which directed GbGGPPS to target to the plastids. Bioinformatic analysis revealed that GbGGPPS was a member of polyprenyltransferases with two highly conserved aspartate-rich motifs like other plant GGPPSs. Phylogenetic tree analysis indicated that plant GGPPSs could be classified into two groups, angiosperm and gymnosperm GGPPSs, while GbGGPPS had closer relationship with gymnosperm plant GGPPSs. Keywords: Geranylgeranyl diphosphate synthase; Ginkgo biloba; Ginkgolide biosynthesis; RACE Database Accession No.: AY371321 Ginkgo biloba is one of the oldest living plant species (Jacobs and Browner, 2000) on the earth and the sole extant representative of the Ginkgoales order (Carrier et al., 1998), which dates back to more than 200 million years. Therefore, G. biloba is often referred to as “a living fossil” that contains many active ingredients, including flavonol and flavone glycosides, diterpene lactones, ginkgolides, sesquiterpenes, iron-based superoxide dismutase, p-hydroxybenzoic acid, ascorbic acid and catechin (Jacobs and Browner, 2000). About 4 billion dollars were spent on botanical medicines in USA in the year 1998 and G. biloba was ranked first among herbal medications sold in health food stores (Brevoort, 1990). Ginkgo leaf extracts have been employed for treating cerebrovascular and cardio- vascular diseases for centuries because ginkgolides are highly specific and potent platelet-activating factor receptor antagonists (Hosford et al., 1990). The commercial ginkgolides are produced merely from G. biloba plants, especially from the ginkgo leaves. The contents of ginkgolides are very low, with about 0.06% ginkgolides in dry leaves from Chinese good-quality ginkgo leaves (Van Beek et al., 1991), in the native ginkgo plant materials. Further- more, the native ginkgo plant materials are limited. Some studies have been made on ginkgo cell and tissue culture with the aim of producing ISSN 1042-5179 print/ISSN 1029-2365 online q 2004 Taylor & Francis Ltd DOI: 10.1080/10425170410001667348 *Corresponding author. Address: State Key Laboratory of Genetic Engineering, Fudan-SJTU-Nottingham Plant Biotechnology R&D Center, School of Life Sciences, Fudan University, Shanghai 200433, People’s Republic of China. Tel.: þ 86-21-65642772. Fax: þ 86-21- 65643552. E-mail: kxtang1@yahoo.com; kxtang1@sohu.com DNA Sequence, April 2004 Vol. 15 (2), pp. 153–158