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目的:探讨甘草β-香树酯醇合成酶(β-amyrin synthase,bAS)编码区SNP与甘草酸含量之间的相关性。方法:采用HPLC法测量80株人工栽培甘草的甘草酸含量,采用SAS 9.0软件将80株甘草按照甘草酸含量极显著水平(P<0.000 1)进行分组;采用RT-PCR技术,扩增出甘草bAS编码区序列,运用DNAman分析软件找出该序列的SNP位点,进而分析该位点与甘草酸含量高低的相关性。结果:bAS基因编码区共有94,254 bp 2个突变位点,在94 bp位点发生G/A转换,为错义突变,导致该位点处甘氨酸/天冬氨酸转换,254 bp处发生C/T转换,为同义突变,根据序列变异将所测样品划分成G-T基因型、A-T基因型、G-C基因型和A/G-C基因型。结论:A-T基因型、G/A-C基因型和G-T基因型和高含量甘草酸形成具有显著的相关性。
Objective: To investigate the correlation between the content of glycyrrhizin and SNP in the coding region of β-amyrin synthase (bAS) of Glycyrrhiza uralensis. Methods: The glycyrrhizic acid content of 80 artificial licorice cultivars was measured by HPLC. The 80 licorice plants were grouped according to the extremely significant levels of glycyrrhizic acid (P <0.0001) using SAS 9.0 software. bAS coding region sequence, using DNAman analysis software to find out the sequence of SNP loci, and then analysis of the site and the level of glycyrrhizin correlation. Results: There were 94,254 bp mutation sites in the bAS gene coding region and a G / A transition at 94 bp, which was a missense mutation, resulting in the glycine / aspartate conversion at this site. The C / T conversion, synonymous mutation, according to the sequence variation of the samples were divided into GT genotype, AT genotype, GC genotype and A / GC genotype. CONCLUSION: There is a significant correlation between A-T genotype, G / A-C genotype and G-T genotype and high content of glycyrrhizin.