Endophytic fungi were isolated from healthy vegetative organs (leaves, roots and stems) of the invasive plant Xanthium strumarium L. harvested from 3 regions of Uzbekistan. This study aimed to characterise the molecular diversity of endophytic fungi associated with this invasive species. Fungal identification was performed using internal transcribed spacer ribosomal DNA (ITS rDNA) sequencing followed by Basic Local Alignment Search Tool (BLAST) analysis against GenBank and MycoBank databases. A total of 361 isolates were grouped into 36 operational taxonomic units (OTUs), representing 20 fungal genera. Leaves harboured the highest proportion of isolates (53.19 %), followed by roots and stems. The genus Alternaria, particularly Alternaria alstroemeriae, was the dominant taxon, while Fusarium species were also frequently detected. Several OTUs exhibited relatively low sequence similarity to reference sequences, indicating the presence of genetically distinct taxa. This study provides the first molecular insight into the diversity of endophytic fungi associated with X. strumarium populations in Uzbekistan and highlights their potential ecological significance.