The effects of different acquisition access periods (AAPs) and inoculation access periods (IAPs) on the transmission efficiency of barley yellow dwarf luteovirus (BYDV) by Rhopalosiphum padi (L.) (Homoptera: Aphididae) after feeding on transgenic or nontransformed wheat, Triticum aestivum L., genotypes were studied. Three wheat genotypes were tested as virus sources: virus-susceptible ‘Lambert’ and ‘Lambert’-derived transgenic lines 103.1J and 126.02, which express the BYDV-PAV coat protein gene. Lower virus titers were measured in BYDV-infected transgenic plants compared with Lambert. No significant differences in transmission efficiency were detected for R. padi after varying IAPs, regardless of genotype. Transmission efficiency increased with an increase in AAP in all genotypes tested. However, AAPs ranging from 6 to 48 h on Lambert resulted in significantly greater transmission efficiency than similar periods on transgenic 103.1J. Maximum transmission efficiency (70%) was observed after a 48-h AAP on Lambert, whereas the same AAP on 103.1J and 126.02 resulted in a significantly lower transmission efficiency (57%). Contrasts were used to compare the rates of transmission and the theoretical maximum transmission percentage among the different wheat genotypes serving as virus sources. Both parameters were significantly different among genotypes, indicating that viral acquisition from each genotype resulted in a unique pattern of virus transmission by R. padi. The lowest rate of virus transmission after an AAP was observed on 103.1J compared with 126.02 or Lambert. This is likely associated with a lower virus titer in 103.1J. This is the first report of transgenic virus resistance in wheat affecting the transmission efficiency of a virus vector.