The Gentile di Puglia sheep breed represents an important component of Southern Italy’s livestock biodiversity, yet its wool traits have not been systematically evaluated for several decades. This study aimed to provide an updated phenotypic assessment of key wool characteristics and to explore the structure of their variability across production contexts. Data were collected from 3089 animals from nine farms over two consecutive years and included fibre diameter, staple length, coefficient of variation in fibre diameter, and clean yield. Descriptive statistics, correlation analysis, principal component analysis, and mixed-effects models were applied to evaluate phenotypic patterns and partition sources of variability. Fibre diameter and staple length showed moderate variability, with most variation occurring within farm-year units, whereas clean yield exhibited a stronger dependence on farm-year conditions. Correlations among traits were generally weak, indicating a high degree of independence. The multivariate analysis revealed a diffuse structure, with no dominant axis explaining most of the variability. Variance component analysis confirmed that production context had a greater influence on clean yield than on fibre-related traits. These results highlight the importance of both conservation and management strategies. The observed variability may provide useful opportunities for future breeding and conservation strategies, while the sensitivity of clean yield to environmental and management factors underscores the need for improved handling practices. Overall, the Gentile di Puglia breed retains a flexible phenotypic profile that supports its conservation and potential valorisation in sustainable production systems.