Guizhou is one of the centers of origin for the tea plant (Camellia sinensis (L.) Kuntze). The location contains highly diverse ancient tea plant germplasms in its Sandu Aquarium Autonomous County. After a prolonged course of continuous evolution, these ancient plants have gained a wealth of genetic diversity. Their resources could be harnessed for the selection and breeding of fine varieties of tea plant, as well as for the effective utilization and protection of germplasm resources. In this study, the specific locus-amplified fragment (SLAF) sequencing method was used to analyze the population structure and conduct a genome-wide association study (GWAS) for the three traits of 125 ancient tea plants in the Sandu County of Guizhou province, China. A total of 807,743 SLAF tags and 9,428,309 population single-nucleotide polymorphism (SNP) tags were obtained. The results of the phylogenetic tree analysis, cluster analysis, and principal component analysis showed that 125 germplasms were clustered into four groups, and the heterozygosity rates for groups I, II, III, and IV, were 0.211, 0.504, 0.144, and 0.192, respectively. Additionally, GWAS analysis suggested that seven candidate genes were related to altitude at the origin of the plants, eight were related to tree shape, and three were associated with leaf color. In this study, we clarified genetic relationships between four ancient tea plant-producing areas in Sandu County and obtained candidate genes related to their development associated with altitude, tree shape, and leaf color. The study provides useful information for tea plant-breeding development and molecular identification.