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Quantitative Analysis of Multiple Components for Quality Evaluation and Four Key Gene Expression Analysis in Different Tissues of Salvia Miltiorrhiza Germplasm

Author: HouShuang
Tutor: BaiZhiChuan；WeiJianHe
School: Southwestern University
Course: Cultivation of medicinal plants
Keywords: Salvia Active ingredient HPLC Simultaneous determination of Clone Tanshinone biosynthetic pathway Key Enzyme Quantitative PCR
CLC: R284.1
Type: Master's thesis
Year: 2010
Downloads: 143
Quote: 1
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Abstract


This thesis consists of two parts: (1) to determine the content of fat-soluble components of Salvia two and three water-soluble components at the same time simple and accurate method of high performance liquid chromatography, using the method of analysis of Salvia 11 clones the content of the active ingredients in the Department of Materials, compared Salvia xylem and phloem, different root diameter thickness as well as the content of the five components of the different root color; (2) the use of quantitative PCR tanshinone biosynthetic pathway SmAACT, SmDXR, SmHMGR, and SmCPS four genes the in Salvia expression in different organs were analyzed. The main research contents: (1) Summary of the chemical composition of Salvia and tanshinone compounds biosynthetic gene Research; (2) establish a HPLC method for simultaneous determination of two kinds of fat-soluble ingredients and three water-soluble components, the method repeatability good precision and stability; (3) from different localities, germplasm Salvia quality; (4) to study to Salvia different parts, different roots thickness and different root color; ⑤ by quantitative PCR SmAACT, SmHMGR SmDXR and SmCPS 4 gene expression studies. The results show that: Salvia clonal material fat-soluble ingredients and water-soluble ingredients are very different, Jiangsu wild (DC003), Wild (DC004) and Henan Yuzhou (DC007) three regions of the sample water-soluble content is relatively high total ; Henan Yuzhou (DC007) samples of fat-soluble component of the total content of the highest data variance analysis showed that the different clones material Danshensu, Dan acid B and tanshinone IIA content reaches significant difference. DC003, DC004, DC007 these three lines can be used as raw material for hybridization to filter out water-soluble and fat-soluble components were higher Salvia germplasm; Salvia different parts of the the different roots thickness and the findings of the different root color: skin Ministry Danshensu acid B, cryptotanshinone, tanshinone IIA were higher than in the xylem; Salvia root diameter is finer, the higher the content of Cryptotanshinone and IIA, salvianolic acid B is the lower, protocatechualdehyde the content of the highest content in the reed head; Salvia root powder color the more closer to red, and the deeper the color, the higher the content of the fat-soluble ingredients, water-soluble component content is higher. In addition, to speculate that SmCPS 4 gene expression results from the tanshinone compounds biosynthetic pathway regulation. To establish a method for the simultaneous determination of three kinds of water-soluble components of Salvia and 2 kinds of fat-soluble ingredients, provides a fast method for multi-component evaluation of Salvia miltiorrhiza quality. The method was applied to different parts of Salvia different germplasm, different root diameter thickness, the active ingredient in a comparative study of the different root color, GAP standardized cultivation and drug companies use Salvia Salvia breeding of new varieties, raw herbs provide a theoretical basis. This article tanshinone biosynthetic pathway of four key enzyme gene expression patterns at the transcriptional level, is conducive to a better understanding of the molecular mechanism of tanshinone biosynthetic pathway, the theoretical basis for Salvia Tanshinone Biosynthesis.

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CLC: > Medicine, health > Chinese Medicine > Of Pharmacy > Traditional Chinese medicine chemical > Chemical analysis and identification
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