Ailanthus altissima, which is recognized as an invasive tree in the Western world, has been widely observed in Japan. To investigate how A. altissima expanded within-population and to new populations within a region, 446 A. altissima trees were sampled from three separate sites (A, B, and C) including 35 distantly positioned patches, with three chloroplast DNA markers and nine nuclear microsatellite markers. We detected 2, 2, and 3 chloroplast haplotypes in sites A, B, and C, respectively. In addition, 271, 40, and 41 nuclear genotypes were detected in sites A, B, and C, respectively. The clonal richness value was 0.85, 0.78, and 0.53 in sites A, B, and C, respectively. Most trees with the same genotypes were distributed in the same patch, indicating that range expansion by asexual reproduction was limited to a maximum of 45 meters. According to autocorrelation analysis, the extent of nonrandom spatial genetic structure was approximately 0-2 km in sites A and C. KINGROUP analyses showed that 812, 74, and 111 nuclear genotype pairs were detected to have kinship in sites A, B, and C, respectively. Most nuclear genotype pairs were detected within the same patches or sites. These results indicate that the number of A. altissima trees gradually increased from seeds, some of which were produced by trees within sites, meaning that this species could regenerate naturally. This shows the need for the future management of A. altissima as an invasive species in Japan.