This study provides a comprehensive examination of the diversity and distribution of micromycete fungi colonizing trees and shrubs in Samarkand, one of the oldest and ecologically significant cities in central Uzbekistan. The research documents a total of 132 micromycete species belonging to 61 genera, revealing a rich fungal diversity in the urban green spaces of the region. Of particular importance is the identification of 32 species that are recorded for the first time in the mycobiota of Uzbekistan, including Phyllosticta albizinae, Cercospora muelleriana, Ramularia philadelphi, Colletotrichum aucubae and Erysiphe euonymicola. These findings indicate both the underexplored nature of urban fungal biodiversity in Central Asia and the ecological significance of Samarkand's dendroflora as a host for diverse fungal communities.The most frequently encountered taxa were members of the genera Camarosporium, Phyllosticta, Pseudocercospora, Diplodia, Erysiphe and Melampsora, which are known to include several phytopathogenic and endophytic species. The checklist compiled in this study includes detailed data on fungal species, their associated host plants and observed distribution patterns, contributing to the foundational knowledge necessary for monitoring plant health and managing urban ecosystems. The vegetation of Samarkand is predominantly composed of mesophytic plant species and demonstrates high floristic diversity. Also, planting many non-native trees and shrubs has created complicated relationships between hosts and pathogens, which adds to the variety of micromycete communities. This study revealed the importance of continued fungal biodiversity assessments in urban environments across Central Asia.