In 2014, disease symptoms of yellowing, foot rot and drying of leaves were observed in vineyards in Hormozgan province, Iran. The goal of the present study was to characterise fungal isolates associated with black foot of grapevines (Vitis spp.) using multi-gene DNA analysis (partial translation elongation factor-1 [tef1], internal transcribed spacers [ITS rDNA] and β-tubulin) and pathogenic characteristics of the isolates from the grapevines. Twenty-five isolates were obtained from diseased plants and identified as Campylocarpon fasciculare (14), Fusarium solani (7) and F. decemcellulare (4) through morphological characteristics. The three DNA regions analysed supported the morphological concept. All fungal isolates were evaluated for their pathogenicity on one-year-old rooted grapevine cultivar Askari in the planthouse. Typical root rot symptoms were observed within 90 days after inoculation. Campylocarpon fasciculare and an unnamed phylogenetic species of FSSC 20 were reported for the first time for Iranian mycoflora, indicating that grapevine vineyards have become the new host plants for F. decemcellulare.