Sharka (Plum pox virus, PPV) severely affects the production of most Prunus species in the areas affected by the disease. In this study, the resistance of 15 Prunus rootstocks to a Dideron type isolate of PPV was evaluated under controlled conditions in an insect-proof greenhouse. After four cycles of study, ÔGF677Õ almond · peach hybrid, ÔMyrobolan 29CÕ plum and ÔL2Õ cherry did not show any symptoms and were ELISA-DASI and RT-PCR negative. These were considered resistant to PPV. The rest of the rootstocks assayed showed symptoms of sharka (confirmed by ELISA-DASI or RT-PCR), although the level of susceptibility was different for each rootstock. ÔGF305Õ peach, ÔPuebla de SotoÕ plum and ÔReal FinoÕ apricot, were highly susceptible to PPV, showing strong sharka symptoms and being ELISA and RT-PCR positive. ÔMarianna 2624Õ plum, ÔAC 9921-07Õ hybrid and ÔCP-2Õ plum showed susceptibility to PPV confirmed by ELISA-DASI and RT-PCR positives. ÔNemaguardÕ and ÔNemaredÕ hybrids, ÔTorinelÕ plum and ÔSTN2Õ hybrid showed an intermediate susceptibility to PPV with slight sharka symptoms and were ELISA-DASI and RT-PCR positive. Whereas, ÔMontclarÕ peach and ÔEvricaÕ hybrid showed moderate resistance to PPV with slight sharka symptoms and were ELISA positive but RT-PCR negative. The results open new possibilities in the search for different sources of resistance to PPV within Prunus.