Transmission of Potato yellow mosaic virus (PYMV) (bipartite begomovirus) to tomato by 50 Bemisia tabaci biotype B individuals was observed when the acquisition access period (AAP) was at least 3 h and the inoculation access period (IAP) at least 30 min. The transmission efficiency increased with the access period to reach 92% transmission after a 48-h IAP and 48-h AAP. The transmission efficiency decreased when whiteflies were fed on PYMV non-host plant between AAP and IAP. According to these results, we inoculated nine Solanum accessions with 50 whiteflies (48-h APP and 48-h IAP) to assess their resistance level. Four of these accessions with various levels of resistance were graft inoculated with PYMV. Although none of the accessions were immune, we observed a high level of resistance to PYMV in Solanum pimpinellifolium LA2187-5 (no symptoms after vector or graft inoculation) and in Solanum chilense LA1969 (no symptoms after vector inoculation and one plant with symptoms after graft inoculation). Inheritance of LA2187-5 resistance, investigated in F1 and F2 populations, appeared to be recessive. Fewer plants were infected by PYMV in S. pimpinellifolium LA1478 after vector inoculation than after graft inoculation. We hypothesised that this was because of vector resistance, which could also be effective against other begomoviruses.