ABSTRACT We demonstrated that only 11 isolates were associated with DNAbeta among 39 Tobacco curly shoot virus (TbCSV)-infected, field-collected samples. An infectious clone of TbCSV-[Y35], an isolate associated with DNAbeta, induced severe upward leaf curling in Nicotiana benthamiana. In the presence of its cognate DNAbeta (TbCSV-[Y35] DNAbeta), the symptom changed to a downward leaf curl. Furthermore, TbCSV-[Y35] alone was able to induce severe symptoms in tobacco and tomato plants, although co-infection with DNAbeta intensified symptom severity in tobacco plants. In contrast to other begomovirus-DNAbeta complexes, the satellite had no effect on the accumulation of TbCSV-[Y35] DNA in systemically infected host plants. The betaC1 mutant caused symptoms comparable to those induced by TbCSV-[Y35] in the absence of DNAbeta. TbCSV-[Y35] can be transmitted between plants by a whitefly vector, regardless of the presence or absence of DNAbeta. For a TbCSV isolate not associated with DNAbeta (TbCSV-[Y1]), systemic infection of N. benthamiana induced symptoms resembling those of TbCSV-[Y35]. Co-infection of TbCSV-[Y1] with TbCSV-[Y35] DNAbeta induced symptoms similar to those following infection by TbCSV-[Y35] and its DNAbeta. This indicates that TbCSV DNAbeta is not necessary for infection but intensifies symptoms in a host-dependent manner. Thus, TbCSV may represent an evolutionary intermediate between the DNAbeta requiring begomoviruses and the truly monopartite begomoviruses. The relevance of these results to our present understanding of the evolution of begomovirus-satellite disease complexes is discussed.