In this study,27 pairs of EST-SSR primers were employed to analyze the genetic diversity and genetic relationship of 100 wild tea plant germplasm resources and 22 cultivars. according to the results,a total of 88 polymorphic bands were amplified with 27 pairs of primers; the variation of effective alleles accounted for 69. 01%; a total of 183 genotypes were detected,with a variation range of 4- 11; averagely 6. 78 genotypes were amplified with each primer pair; Shannon index( I) of 27 primer pairs ranged from 0. 32 to 1. 35,with an average of 0. 88; the observed heterozygosity(0. 52) was basically consistent with the expected heterozygosity(0. 52); the average polymorphism heterozygosity was 0. 48,which was very close to 0. 50; the average Nei’s index was 0. 51,which was higher than0. 50; the average polymorphism information content( PIC) was 0. 52,which was higher than 0. 50,indicating high genetic diversity among wild tea germplasm resources in Yunnan Province. According to the clustering results,based on geographical origins and genetic backgrounds,122 materials were clustered into 14 categories. Dendrogram based on Nei’s genetic distance revealed complex genetic relationships among wild tea germplasm resources in Yunnan Province. This study provided certain reference for subsequent preservation,development and research of wild tea germplasm resources in China. In this study, 27 pairs of EST-SSR primers were used to analyze the genetic diversity and genetic relationship of 100 wild tea plant germplasm resources and 22 cultivars. According to the results, a total of 88 polymorphic bands were amplified with 27 pairs of primers ; the variation of effective alleles accounted for 69.01%; a total of 183 genotypes were detected, with a variation range of 4- 11; averagely 6. 78 genotypes were amplified with each primer pair; Shannon index (I) of 27 primer pairs ranged from 0.32 to 1.35, with an average of 0.88; the observed heterozygosity (0. 52) was substantially consistent with the expected heterozygosity (0. 52); the average polymorphism heterozygosity was 0. 48, which the average Nei’s index was 0. 51, which was higher than 0.50; the average polymorphism information content (PIC) was 0. 52, which was higher than 0.50, indicating high genetic diversity among wild tea germplasm resources in Yunnan Province. According to the clust ering results, based on geographical origins and genetic backgrounds, 122 materials were clustered into 14 categories. Dendrogram based on Nei’s genetic distance revealed complex genetic relationships among wild tea germplasm resources in Yunnan Province. This study provided some reference for subsequent preservation, development and research of wild tea germplasm resources in China.