Abstract: Evaluation of genetic relationship between Zhaoqing local citrus germplasm resources and other citrus cultivars and related genera by simple sequence repeat (SSR) molecular marker technique, the results of present study may provide solid supporting evidence for their classification and breeding.The materials include main citrus cultivars planted in this region (8 samples), other Citrus cultivars (20 samples), Poncirus (1 sample), and Atalantia (1 sample, used as out-group) for PCR amplification with 13 pairs of SSR primers, the amplification product was tested in 1×TBE buffer solution using 1.5% agarose gel electrophoresis. Cluster analysis was performed using UPGMA, and a cluster tree was built using the NTSYSpc2.1 analysis software. Meanwhile, the genetic similarity coefficient was calculated. A total of 13 primer pairs with high polymorphism were selected from the 34 pairs of SSR. The codes of primer sequence with polymorphism are TAA3，CTT01，TAA15，SS18，TAA1，SS16，CMS30，CAC19，CMS24，CSSR036，CMS14，CMS21，CMS20. A total of 83 polymorphic bands were amplified. Each primer has an average of 6.4 polymorphic bands. The average polymorphism is 100%. The clustering results show that the SSR molecular marker technique can reveal genetic diversity and differentiate citrus, Poncitrus, and Atalantia genera from one another. This method is proven effective in identifying citrus species, breeds, and strains in this region. SSR molecular marker technique can effectively differentiate Zhaoqing local citrus germplasm resources . Tested materials showed a closer genetic relationship.