ABSTRACT
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.