ABSTRACT
OBJECTIVE: To construct root-specific promoter of plant expression vector and transformate into Rhodiola hairy root. METHODS: The expression vector pCA-Tob7: UGTR, driven by tobacco root-specific promoter TobRB7, was constructed from pCAM-BIA1301 by substituting the CaMV 35S promoter and GUS gene with TobRB7 and UGTR(a glycosyltransferase gene), respectively. The pCA-Tob7: UGTR vector was introduced into Agrobacterium tumefaciens strain, and the hairy root of Rhodiola were then transformed. RESULTS: Some Kanamycin resistant plants were positive,which indicated that the expression vector had integrated into Rhodiola hairy root genome. CONCLUSION: Root-specific promoter of plant expression vector is constructed and the hairy root of Rhodiola are transformed for future research.
Subject(s)
Genetic Vectors/genetics , Glucosyltransferases/genetics , Nicotiana/genetics , Plants, Genetically Modified/genetics , Promoter Regions, Genetic/genetics , Rhodiola/genetics , Transformation, Genetic , Agrobacterium , Cloning, Molecular , Gene Expression Regulation, Plant , Glucosyltransferases/metabolism , Plant Roots/genetics , Plant Roots/growth & development , Plant Roots/metabolism , Plants, Genetically Modified/metabolism , Rhodiola/metabolism , Rhodiola/microbiology , Nicotiana/enzymologyABSTRACT
Salidroside, the 8-O-ß-D-glucoside of tyrosol, is a novel adaptogenic drug extracted from the medicinal plant Rhodiola sachalinensis A. Bor. Due to the scarcity of R. sachalinensis and its low yield of salidroside, there is great interest in enhancing production of salidroside by biotechnological manipulations. In this study, two putative UDP-glycosyltransferase (UGT) cDNAs, UGT72B14 and UGT74R1, were isolated from roots and cultured cells of methyl jasmonate (MeJA)-treated R. sachalinensis, respectively. The level of sequence identity between their deduced amino acid sequences was ca. 20%. RNA gel-blot analysis established that UGT72B14 transcripts were more abundant in roots, and UGT74R1 was highly expressed in the calli, but not in roots. Functional analysis indicated that recombinant UGT72B14 had the highest level of activity for salidroside production, and that the catalytic efficiency (Vmax/Km) of UGT72B14 was 620% higher than that of UGT74R1. The salidroside contents of the UGT72B14 and UGT74R1 transgenic hairy root lines of R. sachalinensis were also â¼420% and â¼50% higher than the controls, respectively. UGT72B14 transcripts were mainly detected in roots, and UGT72B14 had the highest level of activity for salidroside production in vitro and in vivo.
Subject(s)
Glucosides/biosynthesis , Glycosyltransferases/metabolism , Rhodiola/enzymology , Acetates , Cyclopentanes , Glycosyltransferases/genetics , Oxylipins , Phenols , Phylogeny , Plant Roots/enzymology , Plants, Genetically Modified/enzymology , Recombinant Proteins/metabolism , Rhodiola/geneticsABSTRACT
OBJECTIVE: To compare the chemical constituents of Aetherolea and Alcohol Extraction of purple common perilla of ChangBai Mountain. METHODS: We used water vapor distillation and alcohol extraction method, then analyzed identification by thin-layer chromatography and GC-MS. RESULTS: The main chemical compositions of aetherolea were caryophyllen, caryophyllene oxid, ylangene, 4-(2-Methylcyclohex-1-enyl)-but-2-enal and 3,7-dimethyl-1,6-Octadien-3-ol and the main chemical compositions of alcohol extraction were 10-hendeca-acetylene-1-alcohol, n-hexsdecanoic acid (E)-7,11-dimethyl-3-methylene- 1,6,10-Dodecatriene and so on. CONCLUSION: Other specific compositions including tau. -Muurolol,3H-3a,7-Methanoazulene,2 ,4,5,6,7,8-hexahydro-1,4,9,9-Tetram-ethyl-, [3aR-(3a. alpha, 4. beta 7. alpha)], Astaxanthin and Curan-17-oicacid, 19-acetyl-2,16-didehydro-20-hydroxy-, methyl ester, (19S) have special functions of sterilizing, cancer preventing and immunity improving, which are significantly different from the compositions extracted from other species.