A German isolate of Beet mild yellowing virus (BMYV-IPP) was used for RT-PCR-based construction of the first infectious full-length cDNA clone of the virus (BMYV(fl)). The complete genomic sequence was determined and displayed high similarity to the French isolate BMYV-2ITB. The host range of BMYV(fl) was examined by agroinoculation and aphid transmission. Both methods lead to systemic infections in Beta vulgaris, Nicotiana benthamiana, N. clevelandii, N. hesperis, Capsella bursa-pastoris and Lamium purpureum. Immunological investigation by tissue-print immunoassay (TPIA) of agroinoculated plant tissues revealed only local infections restricted to the agroinoculated mesophyll tissues in some plant species. In Nicotiana glutinosa and N. edwardsonii, BMYV was not found in either the agroinoculated tissue or distant tissues by TPIA. So far, BMYV(fl) agroinoculation did not extend or confine the BMYV host range known from aphid transmission experiments but it did describe new local hosts for BMYV.