Discovering mechanisms of plant–virus–vector interactions is fundamental to understand their ecology and evolution and to apply this knowledge in plant protection. To study the influence of varying Barley yellow dwarf virus (BYDV) transmission efficiencies on host plant preference of Rhopalosiphum padi (L.) clones, we performed host choice experiments with two barley cultivars (BYDV susceptible cv. ‘Rubina’ and BYDV tolerant cv. ‘Vixen’) including healthy and virus-infected plants. For the susceptible barley cultivar ‘Rubina’, aphid clone R07 (high transmission efficiency) preferred BYDV-infected over healthy host plants after 24 h, while clones D10 (medium transmission efficiency) and R09 (low transmission efficiency) preferred neither host. In contrast, BYDV infection of ‘Vixen’ did not affect the plant’s appeal for aphid clone R07. Host plant access, indicated by ingestion and observed by electrical penetration graph technique for a period of 2 h, was facilitated on BYDV-infected cv. ‘Rubina’ for the clones R07 and D10, whereas an opposite effect was observed for the clone R09. For R07 and R09, the difference was not visible after a period of 5 h. As observed earlier for BYDV-infected wheat, enhanced emission of volatile organic compounds associated with virus-induced attraction was detected for BYDV-infected cv. ‘Rubina.’ It is concluded that host plant preference is possibly linked with a high BYDV transmission efficiency as observed for the clone R07, leading to a fitness advantage of this clone as indicated by early increased ingestion. This advantage is not present on BYDV-tolerant genotypes most likely due to the absence of infection symptoms.