【Objective】This study aimed to explore the utilization potential of preserved tea germplasm and breed new elite tea cultivars, and to provide a theoretical and practical references for the breeding of new tea varieties and efficient identification of tea germplasm in Guizhou Province. 【Method】A total of 128 tea germplasm preserved in the resource nursery were used as materials. 6 agronomic traits (including cold resistance, phenological stage, bud and leaf morphology, etc.) and 14 biochemical components (including catechins, alkaloids, theanine, etc.) were systematically determined. Comprehensive evaluation was conducted by combining genetic diversity analysis, correlation analysis, cluster analysis, principal component analysis (PCA), and grey relational analysis (GRA). 【Result】The variation coefficients of 20 trait indices of the 128 tea germplasm ranged from 13.33% to 114.10%, and the genetic diversity indices ranged from 0.92 to 2.08. The average variation coefficient (31.66%) and genetic diversity index (1.63) of agronomic traits were lower than those of biochemical traits (49.80%, 1.86). The length of one bud and three leaves, theogallin, caffeine, epigallocatechin gallate (EGCG), epicatechin gallate (ECG), and theanine contents showed normal distribution, while the kurtosis values of gallocatechin gallate (GCG) and 1,2,6-tri-O-galloyl-β-D-glucose (1,2,6-TGGP) exceeded 20, indicating that the contents of these two components were highly concentrated in the 128 tea germplasm. Correlation analysis showed that the frost injury index was significantly positively correlated with bud and leaf pubescence (P<0.05), and significantly or extremely significantly negatively correlated with bud and leaf color, theobromine, caffeine, and epicatechin (EC) (P<0.01; P<0.05). One-bud-one-leaf stage was significantly or extremely significantly positively correlated with length of one bud with three leaves, young shoot pubescence, caffeine and EGCG contents (P<0.01; P<0.05), and extremely significantly negatively correlated with theogallin, gallic acid, EC, ECG, and theanine contents. Cluster analysis divided the 128 germplasm into 7 clusters, agronomic traits including phenological stages and yield indicators, together with major biochemical components such as caffeine, EGCG and theanine, were the key characteristics for distinguishing germplasm of different groups. PCA extracted 7 principal components with a cumulative contribution rate of 69.35%. Biochemical traits such as EGCG, caffeine, epigallocatechin (EGC), and EC were key indicators affecting F comprehensive score. Grey relational analysis was carried out by constructing an ideal cultivar based on extra-early sprouting, strong cold resistance, and high theanine content. The weighted grey relational degrees between 128 germplasm and the ideal cultivar ranged from 0.51 to 0.73, Notably, GT-GY-05, HJY-F1-55, GT-AS-41, GT-LP-01, and JAL1 exhibited values exceeding 0.7. 【Conclusion】The 128 tea accessions possessed abundant genetic diversity. Overall, biochemical traits showed higher variation coefficients and genetic diversity indices than agronomic traits. Core components including caffeine, EGCG and ECG exhibited relatively high genetic stability, whereas rare secondary metabolites such as methylated catechins and theogallin showed abundant variation, granting them great potential for breeding elite germplasm with unique traits. GT-AS-41, with a high weighted grey relational degree of 0.71, is characterized by extra-early sprouting, highly cold resistance and relatively high theanine content, showing great potential for breeding new high-quality extra-early-sprouting tea cultivars.