title: Production of dammarane-type sapogenins in rice by expressing the dammarenediol-II synthase gene from Panax ginseng C.A. Mey.
abstract: Ginsenosides are the main active ingredients in Chinese medicinal ginseng; 2,3-oxidosqualene is a precursor metabolite to ginsenosides that is present in rice. Because rice lacks a key rate-limiting enzyme (dammarenediol-II synthase, DS), rice cannot synthesize dammarane-type ginsenosides. In this study, the ginseng (Panax ginseng CA Mey.) DS gene (GenBank: AB265170.1) was transformed into rice using agrobacterium, and 64 rice transgenic plants were produced. The Transfer-DNA (T-DNA) insertion sites in homozygous lines of the T2 generation were determined by using high-efficiency thermal asymmetric interlaced PCR (hiTAIL-PCR) and differed in all tested lines. One to two copies of the T-DNA were present in each transformant, and real-time PCR and Western blotting showed that the transformed DS gene could be transcribed and highly expressed. High performance liquid chromatography (HPLC) analysis showed that the dammarane-type sapogenin 20(S)-protopanaxadiol (PPD) content was 0.35-0.59 mg/g dw and the dammarane-type sapogenin 20(S)-protopanaxatriol (PPT) content was 0.23-0.43 mg/g dw in the transgenic rice. LC/MS analysis confirmed production of PPD and PPT. These results indicate that a new "ginseng rice" germplasm containing dammarane-type sapogenins has been successfully developed by transforming the ginseng DS gene into rice.
authors.affiliation: College of Food Science, Fujian Agriculture and Forestry University, 15 Shangxiadian Road, CangShan District, Fuzhou, Fujian 350002, China; Agricultural Product Quality Institute, Fujian Agriculture and Forestry University, 15 Shangxiadian Road, CangShan District, Fuzhou, Fujian 350002, China. Electronic address: hzwfau@163.com.
authors.forename: Zhiwei
authors.initials: Z
authors.lastname: Huang
authors.affiliation: Agricultural Product Quality Institute, Fujian Agriculture and Forestry University, 15 Shangxiadian Road, CangShan District, Fuzhou, Fujian 350002, China. Electronic address: linjc2011@126.com.
authors.forename: Juncheng
authors.initials: J
authors.lastname: Lin
authors.affiliation: Agricultural Product Quality Institute, Fujian Agriculture and Forestry University, 15 Shangxiadian Road, CangShan District, Fuzhou, Fujian 350002, China. Electronic address: chengzuxin@163.com.
authors.forename: Zuxin
authors.initials: Z
authors.lastname: Cheng
authors.affiliation: Agricultural Product Quality Institute, Fujian Agriculture and Forestry University, 15 Shangxiadian Road, CangShan District, Fuzhou, Fujian 350002, China. Electronic address: xmfau@163.com.
authors.forename: Ming
authors.initials: M
authors.lastname: Xu
authors.affiliation: Agricultural Product Quality Institute, Fujian Agriculture and Forestry University, 15 Shangxiadian Road, CangShan District, Fuzhou, Fujian 350002, China. Electronic address: 764900262@qq.com.
authors.forename: Xinying
authors.initials: X
authors.lastname: Huang
authors.affiliation: Agricultural Product Quality Institute, Fujian Agriculture and Forestry University, 15 Shangxiadian Road, CangShan District, Fuzhou, Fujian 350002, China. Electronic address: yangzj41@163.com.
authors.forename: Zhijian
authors.initials: Z
authors.lastname: Yang
authors.affiliation: Agricultural Product Quality Institute, Fujian Agriculture and Forestry University, 15 Shangxiadian Road, CangShan District, Fuzhou, Fujian 350002, China. Electronic address: zjg231@126.com.
authors.forename: Jingui
authors.initials: J
authors.lastname: Zheng
mesh_terms: D060054:Agrobacterium / Q000235:genetics; D019883:Alkyl and Aryl Transferases / Q000235:genetics* / Q000378:metabolism; D036145:Ginsenosides / Q000378:metabolism; D008969:Molecular Sequence Data; D012275:Oryza / Q000235:genetics / Q000378:metabolism*; D005894:Panax / Q000235:genetics* / Q000378:metabolism; D030821:Plants, Genetically Modified / Q000235:genetics / Q000378:metabolism; D017422:Sequence Analysis, DNA; D014315:Triterpenes / Q000378:metabolism*; D000097669:Dammaranes
chemical_list: D019883:Alkyl and Aryl Transferases; D036145:Ginsenosides; D014315:Triterpenes; D000097669:Dammaranes; C514651:dammarenediol-II synthase, Panax ginseng
keywords: Dammarenediol-II synthase; Genetic transformation; Ginsenosides; Rice; Sapogenins