ABSTRACT
To research the correlation between accumulation of triterpenoids and expression of key enzymes genes in triterpenoid biosynthesis of Alisma orientale,the study utilized UPLC-MS/MS method to detect eight triterpenoids content in the tuber of A. orientale from different growth stages,including alisol A,alisol A 24 acetate,alisol B,alisol B 23 acetate,alisol C 23 acetate,alisol F,alisol F 24 acetate and alisol G,and then the Real time quantitative PCR was used to analyze the expression of key enzymes genes HMGR and FPPS in triterpenoid biosynthesis. Correlation analysis showed that there was a significant positive relation between the total growth of these eight triterpenoids and the average relative expression of HMGR and FPPS(HMGR: r = 0. 998,P<0. 01; FPPS: r = 0. 957,P<0. 05),respectively. Therefore,the study preliminarily determined that HMGR and FPPS genes could regulate the biosynthesis of triterpenoids in A. orientale,which laid a foundation for further research on the biosynthesis and regulation mechanism of triterpenoids in A. orientale.
Subject(s)
Alisma , Chemistry , Genetics , Chromatography, Liquid , Geranyltranstransferase , Genetics , Hydroxymethylglutaryl-CoA-Reductases, NADP-dependent , Genetics , Phytochemicals , Plant Extracts , Plant Proteins , Genetics , Plant Tubers , Chemistry , Tandem Mass Spectrometry , TriterpenesABSTRACT
Objective To obtain the transcriptome database and differentially expressed genes of Tetrastigma hemsleyanum Diels et Gilg. by Illumina HiSeq 4000; To provide important molecular information for its molecular biology research. Methods Leaves and roots of Tetrastigma hemsleyanum Diels et Gilg. were chosen as experimental materials to conduct transcriptome sequencing. Then bioinformatics analysis of gene function annotations, metabolic pathways, and microsatellites was performed on the test data. Results 24.13 Gb Clean Data were assembled. Afer assembly steps, 84 433 of T. hemsleyanum Unigene were obtained, and then they were compared in the 7 gene database, and 47 766 annotated information of Unigene was obtained. There were 27 790 annotations in the GO database. The number of differentially expressed genes in the roots, stems and leaves was 4989, of which 3511 were up-regulated and 1478 were down-regulated. The COG database obtained 16 152 homologous sequences of Unigene, which were divided into 25 categories. In the KEGG database, there were 14 511 Unigene obtained the corresponding Ko number, which could be divided into 130 branches of signal metabolism, among which the number of Unigene in the ribosome synthesis pathway was the most, with 1042, and there was only 1 Unigene in the biosynthetic pathway of isoflavones. Conclusion A large number of transcripts of the transcriptome were obtained through splicing, assembling and functional annotation of Tetrastigma hemsleyanum Diels et Gilg., which can provide genomic database resources for molecular biology research of Tetrastigma hemsleyanum Diels et Gilg.
ABSTRACT
Objective: To develop an effective and rapid method for the preparation of 23-acetate alisol B from Alisma orientalis. Methods: The SFE-CO2 extract from A. orientalis was injected into high speed counter current chromatography (HSCCC) directly, and eluted with difierent solvent systems. The crystalline purity was detected by HPLC. The structure of the target compound was identified by UV, IR, MS, and NMR. Results: The solvent system composed of n-hexane-ethylacetate- methanol-water (3∶2∶3∶2) was the best. The lower phase was used as the mobile phase and performed at a flow rate of 2 m/min, while the apparatus rotated at 800 r/min, and detected at 254 nm. The prepared alisol B 23-acetate was identified with infrared spectrometry (IR), mass spectrometry (MS), and nuclear magnetic resonance (NMR) detection, and its purity was 99.8% analyzed by HPLC. Conclusion: The established method is relatively simple, fast, and suitable for the fast isolation and separation of alisol B 23-acetate.