Root rot and vine decline of melon (MRRVD) is a destructive disease complex mainly occurring in semiarid cultivation areas. In the last decade, in the melon producing area of Sardinia (Italy), yield reductions up to 100% were recorded due to the occurrence of MRRVD. The aim of the present study was to undertake a two-years survey of fungal pathogens associated with MRRVD grown as a monocrop in two locations in Central Sardinia, Sinis and Sassu, and to investigate the possible role of soil fungal diversity in the disease development. Melon plants were affected by similar symptoms but colonised by a set of soil-borne fungal pathogens different between surveyed sites and cropping seasons, including Plectosphaerella melonis, P. cucumerina, Fusarium solani, Macrophomina phaseolina and Monosporascus cannonballus. Olpidium bornovanus and O. virulentus were isolated using bait plants and detected by NGS analysis. Bait plants had a general decrease in biomass and yellowing of foliage. The presence of most of the isolated pathogens were also confirmed by the NGS analysis of soil microbiome. Our results confirm that among the fungal complex implicated in the occurrence of RRVD in Sardinia, O. bornovanus along with O. virulentus likely assume a key role in the development of the disease alone and possibly in association with other pathogens. Differences in the pathogenic fungal spectrum here recorded in symptomatic roots may be associated with agricultural practices, soil physicochemical characteristics, and fungal community composition and function profile.