This study assesses the effect of Bt-maize on the distribution of maize viruses. Random surveys were conducted in Spain between 2001 and 2006 to evaluate the occurrence of maize viruses in Bt-maize cultivation areas and in areas where this crop had not been introduced. Maize dwarf mosaic virus (MDMV) was the predominant virus in Bt-areas, and Maize rough dwarf virus (MRDV) was the most predominant one in non-Bt-areas, with MRDV an emergent virus in both types of areas. A decline in the occurrence of MDMV and an increase in that of Sugarcane mosaic virus was observed in Bt-areas. Additionally, data obtained over 6 years in experimental fields showed non-significant differences between the infection rates exhibited by two generations of Bt varieties and the non-transformed isogenics varieties for any of the viruses. Our data suggest that differences in virus distribution are linked to the genetic background of the maize varieties and the distribution of virus reservoirs rather than to Bt-maize cultivation.