한빛사 논문
Jin-Wook Seoa, Jae-Hun Jeongb, Cha-Gyun Shina, Seog-Cho Loc, Seong-Soo Hanc, Ki-Won Yud, Emiko Haradae,1, Jeong-Yeon Hane, Yong-Eui Choie,*
aDepartment of Biotechnology, Chung-Ang University, Ansung, Korea
bPlant Biotechnology Division, Korea Forest Research Institute, Suwon, Korea
cDivision of Bio-Environmental Science, Wonkwang University, Iksan, Korea
dResearch Center for the Development of Advanced Horticultural Technology, Chungbuk National University, Chungbuk, Korea
eDivision of Forest Resources, College of Forest Sciences, Kangwon National University, Chunchon, Korea
*Corresponding author
1Present address: Leibriz-Institut für Pflanzenbiochemie, Weinberg 3, 06120 Halle/Saalle, Germany.
Abstract
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 Agrobacterium-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 μM 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 (β-sitosterol and stigmasterol) as well as triterpene saponins (ciwujianosides B (1), C1 (2), C2 (3), C3 (4), C4 (5), D1 (6) and D2 (7)) levels in transgenic E. senticosus 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.
Keywords
Eleutherococcus senticosus; Siberian ginseng; Araliaceae; Ciwujianoside; Genetic transformation; Squalene synthase; Triterpene biosynthesis
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