Evolution of Tomato yellow leaf curl virus–Mild (TYLCV-Mld[RE]) (family Geminiviridae, genus Begomovirus) was monitored in La Réunion island from its first upsurge in 1997 until 2004. Two genome fragments, one comprising partial C4 and C1 open reading frames (ORFs), and the other comprising part of the V1 and V2 ORFs and part of the intergenic region were sequenced in 111 isolates. The very low initial diversity of TYLCV-Mld[RE] in La Réunion was followed by a quasi-linear increase in genetic diversity across years. In addition, the population effective size of TYLCV-Mld[RE] has undergone a sudden increase from 2001 to 2004, which is consistent with a founder effect due to the introduction of a small number of virus individuals in an insular environment. Surprisingly, one nucleotide substitution introducing a premature stop codon in the C4 ORF was observed in an increasing number of isolates in the population of TYLCV-Mld[RE] over time, contrasting with the other substitutions which were observed at low frequencies. This substitution which shortens the C4 protein by four amino acids may therefore have been selected during TYLCV-Mld[RE] evolution.