目的 采用SCoT分子标记方法对茶枝柑及近缘种进行遗传多态性分析。方法 使用正交设计方法，对Mg²⁺、dNTPs、ExTaq DNA聚合酶、引物浓度及模板DNA用量5个因素进行筛选，筛选适合茶枝柑及近缘种的SCoT-PCR反应体系，同时通过梯度温度筛选最佳退火温度，并验证其多态性引物。结果 得到适于茶枝柑的SCoT标记PCR反应体系，最终筛选出12条清晰、条带丰富的作为茶枝柑及近缘种SCoT分子标记的引物，同时通过NTSYS软件分析得到茶枝柑及近缘种的相似系数表和UPGMA聚类树。结论 运用3个不同地域柑橘基因组DNA对优化的SCoT-PCR反应体系进行验证，获得了多态性丰富、条带清晰的扩增图谱，表明茶枝柑SCoT分子标记技术体系稳定可靠。而聚类分析结果能够直观、科学地从分子角度对茶枝柑及近缘种13种材料进行初步区分。

Objective To carry out genetic polymorphism analysis of Citrus reticulata Blanco cv. chachiensis Tanaka and its relatives by using SCoT molecular marker method.Methods Five factors of Mg²⁺, dNTPs, TaqDNA polymerase, primer, and template DNA concentration were used to screen the suitable SCoT-PCR reaction system for C. reticulata and its relatives by the method of orthogonal design. The optimum annealing temperature was screened by gradient temperature, and its polymorphic primers were verified.Results A total of 12 clear and rich bands were finally screened out as the primers of SCoT molecular marker for C. reticulata and its relatives in the optimized PCR reaction system, and the genetic distance and the UPGMA clustering tree of C. reticulata and its relatives were got by NTSYS software analysis.Conclusion The optimized SCoT-PCR reaction system was validated by using three different places of citrus genomic DNA to obtain the polymorphism and the clear amplified bands, which showed that the SCoT molecular marker system of Citrus reticulata is stable and reliable. The results of cluster analysis were able to make a preliminary separation of 13 kinds of materials scientifically and intuitively in molecule level.

广东省教育厅高校重点实验室滚动支持项目"春砂仁等十大广药DNA条形码标准化研究"（2013CXZDA011）；国家公益性行业科研专项（201407002）