Grapevine trunk disease pathogens, and specifically Petri disease pathogens, can be spread by planting infected plants. Due to the increasing incidence of Petri disease and other young grapevine declines reported lately in Spain, a sampling of plants used before for new vineyards were carried out in 2002 and 2004. A total number of 208 plants (grafted and non grafted) were collected, of which 94 plants (45.2%) were infected with at least one of the following pathogens Phaeomoniella chlamydospora, and species of Phaeoacremonium, Botryosphaeria, Cylindrocarpon, and Phomopsis. Species of the genera Phaeoacremonium and Botryosphaeria isolated in 2004 were identified using morphological and molecular characters. Species of Phaeoacremonium identified were P. aleophilum and P. parasiticum; and those of Botryosphaeria were B. obtusa, B. dothidea and B. parva. This is the first report of P. parasiticum and B. parva occurring on grapevines in Spain. Distribution of pathogens within the plants was studied in 2004. Phaeomoniella chlamydospora was not detected in the graft union of any plant; however, species of Botryosphaeria and Phomopsis were detected along the plant, but mainly in the graft union; Phaeoacremonium aleophilum was detected along the grafted plants, but not in rooted rootstocks. The results suggest that infected plants used for new plantings in Spain are an important source of primary inoculum of the pathogens associated with grapevine trunk diseases in the field. © Springer 2006.