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目的:揭示甘草3-羟基-3-甲基戊二酰辅酶A还原酶(HMGR)基因的多态性,并阐述HMGR基因多态性与甘草酸含量的相关性。方法:以甘草酸含量差异显著的甘草个体为材料,通过RT-PCR法扩增其HMGR基因编码区;与载体pMD19-T连接后克隆测序;通过多重比对进行HMGR基因多态性分析,及与甘草酸含量之间的关联分析。结果:HMGR基因编码区多态性包括整码突变、碱基置换突变、拷贝数多态性及等位基因杂合;HMGR编码氨基酸序列中存在以下4种类型变异,即-HSL,-HSV,GALLV,GALSV,其中-HSV型为甘草中普遍存在类型,-HSL型为甘草酸低含量甘草的特有类型,GALSV型为甘草酸高含量甘草特有类型。结论:甘草HMGR基因编码区具有丰富的多态性,和甘草酸含量之间具有相关性。
Objective: To reveal the polymorphism of 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMGR) in Glycyrrhiza uralensis and to elucidate the relationship between HMGR gene polymorphism and glycyrrhizin content. Methods: The Glycyrrhiza glabra individuals with significant differences in glycyrrhizic acid content were used as materials. The coding region of HMGR gene was amplified by RT-PCR, cloned into pMD19-T vector and sequenced. The polymorphism of HMGR gene was analyzed by multiple alignment and And glycyrrhizic acid content of the correlation analysis. Results: The HMGR gene coding region polymorphism including whole-coding mutation, base substitution mutation, copy number polymorphism and allelic heterozygosity; HMGR encoded amino acid sequence of the following four types of variation, namely -HSL, -HSV, GALLV, GALSV, of which-HSV type is commonly found in licorice, -HSL type is licorice root licorice low-licorice specific type, GALSV type is licorice root high-licorice licorice-specific type. CONCLUSION: The coding region of Glycyrrhiza uralensis HMGR gene is rich in polymorphism and has a correlation with glycyrrhizic acid content.