Phomopsis cane and leaf spot (PCLS) is an important disease of grapevines, which is mainly caused by Diaporthe ampelina. Dispersal dynamics of D. ampelina spores were investigated in two vineyards, one in North Italy and one in Montenegro, by using spore samplers that collected alpha and beta conidia from rain water running off from PCLS-affected canes. The canes were collected from each vineyard, deployed and overwintered in the corresponding vineyards. In each of three years (2016, 2017, and 2018), conidial dispersal was investigated during one (Montenegro) or two (Italy) growing seasons following the deployment of the PCLS-affected canes. In the first growing season following cane deployment in both vineyards, alpha conidia were mostly found in runoff water after grapevine bud break, especially in April and May, and beta conidia were regularly found in numbers comparable to alpha conidia, most frequently from June to September. In Italy, high numbers of alpha and beta conidia were also collected during the second growing season following cane deployment. The dispersal dynamics of alpha conidia over time were described by a Gompertz equation using hydrothermal time (i.e., the accumulated effect of temperature on the maturation rate of pycnidia on days in which the number of hours of wetness was ≥ 6 or 9 h), with R2 and concordance correlation coefficient >0.9. Rain (≥ 0.2 mm) was a good predictor of conidial dispersal, with an overall accuracy of 0.97. These results increase our understanding of D. ampelina spore dispersal and should be integrated into warning systems for PCLS management.