目的 通过对佛手RAPD反应的优化,分析其遗传多态性,并对 3个品种作出了分子水平上的鉴定。方法RAPD反应在以下条件下进行 :10～ 10 0 ng/μL模板 DNA,3.0～ 5 .0 mmol/ L Mg Cl₂,2 0 0μmol/ LdNTP,0 .2μmol/ L 10 - mer Prim er,lunit Taq Polym Erase/ 2 5 μL 反应体积。结果 从 38个引物中挑出 12个有效稳定的引物。用距离系数 UPGMA法聚类分析产生了树状图。根据 RAPD分析及形态学观察,品种白花青皮与白花白皮的亲缘关系较近。不同的品种RAPD反应有各自独特的扩增带型。结论 本方法简单、高效,适宜用于中药佛手的遗传多态性和品种分子鉴定。

Object To analyse genetic polymorphism in DNA level of three species and determine them in molecular level by using of the optimization of Citrus medica cv. sarcodactylis (Noot.) Swingle RAPD reaction.Methods RAPD reactions were performed under the following conditions:template DNA 10～100 ng/μL, 3.0～5.0 mmol/L MgCl 2, 200 μmol/L dNTP,0.2 μmol/L 10 mer Primer, lunit Taq Polymerase/25 μL reaction volume.Results Twelve efficient and stable primers were selected from 38 primers. Phenograms were constructed by using genetic distance UPGMA method. Based on the results of the RAPD analysis and on a review of morphological characterisitic, C. medica cv. sarcodactylis species BAIHUAQINGPI could be used as BAIHUABAIPI in three species, there were some special amplified bands in every RAPD reactions from different species. Conclusion This method could serve as an easy and highly efficient method for C. medica cv. sarcodactylis genetic polymorphism and molecular identification of species.

浙江省自然科学基金项目资助(301026);浙江省大型仪器测试基金项目资助(00070)