Journal
PHYTOCHEMISTRY
Volume 66, Issue 8, Pages 869-877Publisher
PERGAMON-ELSEVIER SCIENCE LTD
DOI: 10.1016/j.phytochem.2005.02.016
Keywords
Eleutherococcus senticosus; Siberian ginseng; Araliaceae; Ciwujianoside; genetic transformation; squalene synthase; triterpene biosynthesis
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Squalene synthase (SS) catalyzes the first committed step in sterol and triterpenoid biosynthesis. Transgenic Eleutherococcus senticosus Rupr. and Maxim. plants were generated by introducing an SS-encoding gene derived from Panax ginseng (PgSS1) together with genes expressing hygromycin phosphotransferase and green fluorescent protein (GFP) through Agrobacterium-mediated transformation. Early globular embryo clusters developing from the embryogenic callus were used for Agrobacteritan-mediated transformation. Transformants were selected on Murashige Skoog medium containing 25 mg/L hygromycin. Hygromycin-resistant somatic embryos developed into plants after the cotyledonary embryos were treated with 14.4 mu M gibberellic acid. Transformation was confirmed by polymerase chain reaction, Southern, and GFP analyses. The SS enzyme activity of the transgenic plants was up to 3-fold higher than that of wild-type plants. In addition, GC-MS and HPLC analysis revealed that phytosterols (P-sitosterol and stigmasterol) as well as triterpene saponins (ciwujianosides B (1), C-1 (2), C-2 (3), C-3 (4), C-4 (5), D-1 (6) and D-2 (7)) levels in transgenic E senticostis were increased by 2- to 2.5-fold. These results suggest that the metabolic engineering of E. senticosus to enhance production of phytosterols and triterpenoids by introducing the PgSS1 gene was successfully achieved by Agrobacterium-mediated genetic transformation. (c) 2005 Elsevier Ltd. All rights reserved.
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