Several fungi were recovered from velvetleaf plants (Abutilon theophrastni) in eastern Croatia. During early autumn 2002, velvetleaf plants with canker symptoms were collected from sugarbeet and soybean fields. Using standard phytopathological methods (culturing in a moist chamber and isolation on acidified potato dextrose agar at 25°C, with a 12/12 h light/dark regime), a Phomopsis sp. was isolated from diseased stem tissue. The cultural characteristics, pattern of stromata, the formation of only alpha conidia, biometrical comparisons of pycnidia and conidia and prominent pycnidial beaks, identified these two isolates from velvetleaf as Phomopsis longicolla (Hobbs et al., 1985). To confirm the morphological identification of isolates, DNA was extracted from monoconidial cultures and the ITS regions of the rDNA were amplified with universal primers ITS5 and ITS4, and digested with AluI, RseI and MseI restriction enzymes (Riccioni et al., 1998; Riccioni et al., 2003). All isolates showed the characteristic electrophoretic profiles of P. longicolla. The amplified ITS products were also sequenced (M-MEDICAL, Genenco, Rome, Italy) and subsequent searches of the GenBank sequence database revealed that the Croatian isolates from velvetleaf were identical to the sequences of P. longicolla isolates from soybean. Pathogenicity testing was done according to Li et al. (2001). Mean stem lesion lengths caused by the velvetleaf and soybean isolates were 16 and 20·6 mm, respectively, on soybean stems, and 12 and 22 mm, respectively, on velvetleaf stems. Phomopsis longicolla was reisolated from the stem lesions of the inoculated plants. The fungus is a very harmful seed pathogen and for this reason it is of great importance to recognize possible host plants which could be a source of inoculum. A previous paper (Li et al., 2001) reported velvetleaf as a host of P. longicolla in the USA, but this is the first report from Europe.