不同产地柴胡的皂苷含量和ITS序列比较

Comparison of ITS Sequence and Saponin Content of Bupleuri Radix from Different Producing Areas

目的:比较不同产地柴胡药材的皂苷含量和内转录间隔区(ITS)序列,从有效成分含量和遗传基础2个层面为柴胡药材生产和利用提供借鉴。方法:产地收集柴胡药材,采用高效液相色谱法(HPLC)测定样品中柴胡皂苷a、柴胡皂苷c、柴胡皂苷d含量;提取DNA,采用聚合酶链式反应(PCR)扩增ITS序列并比较分析序列差异。结果:各产地柴胡有效成分含量差异较大,柴胡皂苷a、柴胡皂苷c、柴胡皂苷d总质量分数最高可达4.165%,最低为0.118%。不同产地柴胡种质依据ITS序列可分为三大类,其中第Ⅱ类产于甘肃省,第Ⅲ类产于四川省、山西省和甘肃省。第Ⅲ类含有的柴胡样品较多,又可分a和b类,b类又可分为i、ii、iii类,含量较高的样品主要存在于iii小类,主要来源于甘肃、山西。结论:尽管柴胡自然分布广、生产适应性强,但从成分含量和遗传差异角度分析,有必要对柴胡的种植品种和产地进行规范。

Objective:This study aims to compare the saponin content and internal transcribed spacer(ITS)of Bupleuri Radix from different producing areas,which is expected to serve as a reference for the production and identification of this medicinal material based on effective component content and genetic basis.Methods:Bupleuri Radix was collected from different areas and the content of saikosaponin a,c,and d was determined by high performance liquid chromatography(HPLC).The DNA of the samples was extracted,followed by polymerase chain reaction(PCR)amplification of the ITS sequences and analysis of the sequences.Results:The content of effective components in Bupleuri Radix was remarkably different among the producing areas,as the total mass fractions of saikosaponins a,c,and d fluctuated from 4.165%to 0.118%.According to the ITS sequence-based NJ tree,all samples were classified into three categories.CategoryⅡwere from Gansu and categoryⅢfrom Sichuan,Shanxi,and Gansu.CategoryⅢwas further divided into subcategories a and b and the latter was then divided into groups i,ii,and iii.Samples with high content of the effective components were mainly in group iii,which were mainly from Gansu and Shanxi.Conclusion:Although Bupleuri Radix has wide natural habitats and strong adaptability,it is necessary to standardize the varieties and producing areas according to the component content and genetic differences.