Plants have a variety of fungal communities on their surfaces (exophytic) and within their tissues (endophytic). They are essential for plant health and the environment. There is an unexplored diversity of fungal associations with medicinal plants. Catharanthus roseus, a medicinal plant, plays a critical role in global health care. A total of 153 fungal isolates were isolated from thirty leaves of the C. roseus plant, including 11 endophytic and 142 exophytic isolates. Based on morphological characteristics, the isolates were assigned to six endophytic fungi genera and 11 species, as well as 12 exophytic fungi genera and 33 species. Endophytes have significantly less fungal diversity compared to epiphytes. By sequencing the ITS region of rDNA, five epiphytic and three endophytic fungal species were identified molecularly. The isolates were Aspergillus caespitosus, A. elegans, A. flavus, A. nidulans, Penicillium chrysogenum, P. commune and Talaromyces purpurogenus. Aspergillus was the most abundant fungal genus isolated, with four endophytic species out of eleven and nine exophytic species out of thirty-three. A. niger2 was the most common exophytic species, with 28 isolates, an 18.6% colonization frequency, and a relative frequency of 19.7%. In addition, A. caespitosus was the most common endophyte, accounting for 1.3% of the CF and 18.18% of the RF. In this study, we aim to compare epiphytes and endophytes diversity that performed poorly on medicinal plants. This discovery can provide new perspectives and opportunities for compounds and mycotoxins with medicinal value.