Robinia pseudoacacia has become invasively naturalized in Japan. We investigated the role of sexual reproduction in the development of R. pseudoacacia riparian forests along the Chikumagawa River in Japan, by using five chloroplast (cpSSR) and seven nuclear (nSSR) markers. We identified eight chloroplast haplotypes and 147 nuclear genotypes from 619 R. pseudoacacia trees sampled in three plots (Plots A, B, and C) and along two line transects (Lines D and E). CpSSR analyses showed that multiple maternal lines were distributed along the river, and that some haplotypes from different populations overlapped. In addition, while Plots A and B were separated by a short distance, only these two plots exhibited genetic differentiation in the haplotypes. In the nSSR analysis, all pairwise F ST values among the three plots were significantly different from zero. Kinship analysis based on nSSR markers revealed that kinship connected many individuals to another individual from the same plot. These results indicate that seed dispersal near to mother trees contributes to the fine-scale genetic structure of R. pseudoacacia riparian forests. Our results indicate that sexual reproduction, in addition to asexual reproduction, is a major contributor to the fine-scale formation of R. pseudoacacia forests.