Abstract
In Panax ginseng, both ginsenosides and phytosterols are derived from the precursor 2,3-oxidosqualene. The enzymes cycloartenol synthase (CS) and dammarenediol synthase (DS) are responsible for ginsenoside and phytosterol biosynthesis, respectively. Here, we report on enhanced ginsenoside levels detected in P. ginseng hairy roots, following Agrobacterium rhizogenes transformation, by antisense suppression of the CS gene. Several antisense hairy root transgenic lines were recovered exhibiting markedly reduced levels of the CS transcript and corresponding CS enzymatic activity relative to control. Phytosterol levels in tested antisense transgenic lines were ∼50% lower than those of control, whereas 2,3-oxidosqualene contents in tested antisense lines were higher than those in control lines during the early period of root growth; however, these levels were comparable in both groups at a later stage of root growth. Ginsenoside levels were comparable in both groups during the early stage of root growth; however, the final total ginsenoside contents in antisense lines were about 50–100% higher than those found in control roots. Interestingly, the DS enzyme activity of antisense-CS lines was higher than that of controls. It is proposed that in P. ginseng, the regulation of CS can control the metabolic flux from 2,3-oxidosqualene to both phytosterol and ginsenoside.
Similar content being viewed by others
References
Attele AS, Zhou YP, Xie JT, Wu JA (2002) Antidiabetic effects of Panax ginseng berry extract and the identification of an effective component. Diabetes 51:1851–1858 doi:10.2337/diabetes.51.6.1851
Brian M, Carl AM, Aideen HO (2007) Compositions with increased phytosterol levels obtained from plants with decreased triterpene saponin levels. US patent 20070107081, 05/10/2007.
Chang YS, Seo EK, Gyllenhaal C, Block KI (2003) Panax ginseng: a role in cancer therapy? Integr Cancer Ther 2:13–33 doi:10.1177/1534735403251167
Cheng Y, Shen LH, Zhang JT (2005) Anti-amnestic and anti-aging effects of ginsenoside Rg1 and Rb1 and its mechanism of action. Acta Pharmacol Sin 26:143–149 doi:10.1111/j.1745-7254.2005.00034.x
Flores-Sanchez IJ, Ortega-Lopez J, Montes-Horcasitas MC, Ramos-Valdivia AC (2002) Biosynthesis of sterols and triterpenes in cell suspension cultures of Uncaria tomentosa. Plant Cell Physiol 43:1502–1509 doi:10.1093/pcp/pcf181
Furuya T, Yoshikawa T, Ishii T, Kajii K (1983a) Regulation of saponin production in callus cultures of Panax ginseng. Planta Med 47:200–204 doi:10.1055/s-2007-969985
Furuya T, Yoshikawa T, Orihara Y, Oda H (1983b) Saponin production in cell suspension cultures of Panax ginseng. Planta Med 48:83–87 doi:10.1055/s-2007-969892
Giri A, Narasu ML (2000) Transgenic hairy roots: recent trends and applications. Biotechnol Adv 18:1–22 doi:10.1016/S0734-9750(99)00016-6
Guillon S, Tremouillaux-Guiller J, Pati PK, Rideau M, Gantet P (2006) Hairy root research: recent scenario and exciting prospects. Curr Opin Plant Biol 9:341–346 doi:10.1016/j.pbi.2006.03.008
Han JY, Kwon YS, Yang DC, Jung YR, Choi YE (2006) Expression and RNA interference-induced silencing of the dammarenediol synthase gene in Panax ginseng. Plant Cell Physiol 47:1653–1662 doi:10.1093/pcp/pcl032
Haralampidis K, Trojanowska M, Osbourn AE (2002) Biosynthesis of triterpenoid saponins in plants. Adv Biochem Eng Biotechnol 75:31–49
Hong MF, Kong YJ (2000) Determination of total saponin content in Panax ginseng and notoginseng by UV spectrometer. Chin J Mod Appl Pharm 17:51–52
Hong SG, Lee KH, Kwak J, Bae KS (2006) Diversity of yeasts associated with Panax ginseng. J Microbiol 44:674–679
Kushiro T, Shibuya M, Ebizuka Y (1998) Amyrin synthase: cloning of oxidosqualene cyclase that catalyzes the formation of the most popular triterpene among higher plants. Eur J Biochem 256:238–44 doi:10.1046/j.1432-1327.1998.2560238.x
Lee MH, Jeong JH, Seo JW, Shin CG, Kim YS, In JG, Yang DC, Yi JS, Choi YE (2004) Enhanced triterpene and phytosterol biosynthesis in Panax ginseng overexpressing squalene synthase gene. Plant Cell Physiol 45:976–984 doi:10.1093/pcp/pch126
Liang Y, Zhao S (2008) Progress in understanding of ginsenoside biosynthesis. Plant Biol Stuttg 10:415–421 doi:10.1111/j.1438-8677.2008.00064.x
Liu WK, Xu SX, Che CT (2000) Anti-proliferative effect of ginseng saponins on human prostate cancer cell line. Life Sci 67:1297–1306
Mallol A, Cusido RM, Palazon J, Bonfill M, Morales C, Pinol MT (2001) Ginsenoside production in different phenotypes of Panax ginseng transformed roots. Phytochemistry 57:365–371 doi:10.1016/S0031-9422(01)00062-0
Seo JW, Jeong JH, Shin CG, Lo SC, Han SS, Yu KW, Harada E, Han JY, Choi YE (2005) Overexpression of squalene synthase in Eleutherococcus senticosus increases phytosterol and triterpene accumulation. Phytochemistry 66:869–877 doi:10.1016/j.phytochem.2005.02.016
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Liang, Y., Zhao, S. & Zhang, X. Antisense Suppression of Cycloartenol Synthase Results in Elevated Ginsenoside Levels in Panax ginseng Hairy Roots. Plant Mol Biol Rep 27, 298–304 (2009). https://doi.org/10.1007/s11105-008-0087-7
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11105-008-0087-7