In order to understand the breeding status of cotton varieties in Yangtze River region, 100 upland cotton of the national regis-tered varieties breeding from year of 2001 to 2023 were collected. Phenotypic identification was conducted for major agronomic traits, yield-related traits and fiber quality traits. Clustering analysis, correlation analysis, principal component analysis, and comprehensive evaluation methods were employed to study 12 traits. The results revealed a variation coefficient range of 3.06% to 24.37% for each trait, and a genetic diversity index between 0.42 and 2.05. Notably, boll number exhibited the highest variation coefficient, while boll weight displayed the highest genetic diversity index. The results of cluster analysis showed that the tested varieties were divided into three groups, representing the varieties with poor yield and quality performance, the varieties with low yield and high quality, and the varieties with high yield and low quality. Correlation analysis indicated a significant positive correlation between the growth period and most yield-related traits, negatively correlated with fiber length and uniformity, and positively correlated with Micronaire value. Principal component analysis revealed that the cumulative contribution rate of the first three principal components reached 70.23%, highlighting boll number, fiber strength, and fiber length as major factors contributing to phenotypic variation in cotton. Comprehensive evaluation scores ranged from 0.48 to 2.88, identifying 10 varieties with outstanding overall performance. The findings contribute valuable insights into the genetic improvement of cotton, offering both theoretical and practical significance. This study provides a reference for the development of new cotton varieties with improved traits.