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Pueraria medicinal components content of geographic variation genotyping study

Author: YuZhiKui
Tutor: LiuChunSheng
School: Beijing University of Traditional Chinese Medicine
Course: Microbial and Biochemical Pharmacy
Keywords: Pueraria Barcode Genotyping Medicinal components
CLC: R284
Type: Master's thesis
Year: 2009
Downloads: 225
Quote: 0
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Abstract


Pueraria as one of the commonly used in clinical bulk herbs and their medicinal components puerarin has high medicinal value, but under different geographical environment Pueraria medicinal components were quite different, Pueraria efficacy ingredients variability is affected by genetic variation The impact is not yet clear. Therefore, this article Pueraria genotyping studies the combined the Pueraria efficacy ingredients be evaluated, and thus provide the basis for for Pueraria excellent breeding on the genetic level. Genotyping study Pueraria germplasm resources, combined with the medicinal ingredient content of its evaluation, the paper carried out the following work: ① of 24 copies of Pueraria samples collected from eight different habitats, determination of the four DNA barcode gene sequence, Pueraria genotyping studies; and ② measured puerarin content of total flavonoids in Pueraria root and shoot of its pharmacodynamic ingredients geographic variation; and ③ revealed Pueraria Germplasm resource genotype combined the pharmacodynamic ingredients typing results were evaluated screening superior genotypes to Pueraria Germplasm Resources preferred genotype basis. The main conclusions are as follows:. Pueraria ITS sequence variation, more genotypes using the pharmacodynamic ingredients evaluation results showed that genotype influence Pueraria puerarin content in one of the reasons. Eight localities in this study using ITS sequence analysis of 24 the Pueraria samples, genotyping. Pueraria ITS and 5.8S sequence, ITS full sequence, ITS1 sequences, ITS2 sequence were sequence variation analysis results show that 12 samples are common genotype; another 12 samples ITS sequences to multiple sites variation. The these variants information can be divided into 24 parts Pueraria samples experiment 13 genotypes ITS sequences Pueraria genotyping using the pharmacodynamic ingredients evaluation results showed that genotype is affect puerariae puerarin content one of the reasons. Chloroplast genes trnL intron sequence variation, only one genotype. Use of the trnL Intron sequence analysis of eight localities in this study were 24 Pueraria samples, DNA barcode genotyping. Found Pueraria trnL intron sequence variation sites, only one genotype. Use of the chloroplast gene psbK-psbI sequence variation results can Pueraria sample experiment is divided into two genotypes. The sequence of psbK-psbI 24 Pueraria sample analysis of eight localities in the present study, the DNA barcode genotyping. Having two sites in the sample of the measured Pueraria mutation information, wherein there are 16 samples in at nucleotide 249 and 301 bases at T, 8 samples in the two bases at G, which were named \Chloroplast genes trnH-psbA sequence variation results to available the Pueraria samples in the experiment is divided into two genotypes. TrnH-psbA sequence analysis of eight localities in this study were 24 Pueraria samples, DNA barcode genotyping. One mutation while 10 bp deletion mutation, which has the Pueraria samples measured in 9 samples in the first 45 bases at A, 15 samples in the 181-191 base at 9 samples \5. Puerarin total flavonoids of medicinal components without significant change, the puerarin content has significant geographic differences. UV method for determination of total flavonoids in a different origin Pueraria root, content between 9.20 to 9.65%, the relative standard deviation was 0.71%, the different origin of Pueraria total flavonoid content was not significantly different. HPLC method for the determination of different origin Pueraria root puerarin content changes between 0.35 to 8.24%, RSD value of 55%, a significant difference; exclude factors of kudzu growing years, determination of kudzu leaves and stems of different origin Pueraria The prime content changes between 0.83 to 4.47% RSD is 60%, the results show that the difference was very significant. 6. Pueraria different genotypes of total flavonoids in no significant differences, differences in puerarin content. Pueraria lobata flavone content determined by UV, HPLC method to study the medicinal components puerarin content, Pueraria genotyping results were evaluated, the results show that no significant changes in the total flavonoid content of the different genotypes, puerarin content differences . Puerarin content in the the genotyping of psbK-psbI findings, the GG type \4.55%, an average of 2.18% in the puerarin content in the stems and leaves. A non-deletion \average of 4.49%, an average of 2.18% in the puerarin content in the stems and leaves. Thereby filter out the quality Pueraria genotype, i.e. the gene psbK-psbI \Pueraria chloroplast gene psbK-psbI and trnH-psbA genotyping and origin are closely related. Using psbK-psbI sequence genotyping results found that the nucleotide site variation and geographical origin. Liaoning Qianshan, Huairou, Beijing, Hebei Yixian, Hebei and Henan Romer \boundaries, and south of the \Shaanxi Pingli sample \TrnH-psbA sequence genotyping study results found that the nucleotide site variation and geographical origin. Liaoning Qianshan, Huairou, Beijing, Hebei Yixian, Hebei and Henan Romer T deletion, Pinglu, Shanxi, Shaanxi Pingli, Shaanxi Xunyang \Shanxi Pinglu Lushi boundaries south \In summary, Pueraria Germplasm Resources genotyping studies from different areas in this article, the use of total flavonoids and puerarin root, stem and leaf content of genotype evaluation, concluded clarify quality medicinal quality mechanism gene; screening Gegen superior varieties guidance Pueraria cultivation, to lay the foundation to improve the quality of TCM.

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