Japanese Climbing Fern (Lygodium japonicum) is a vine native to the open forests of eastern Asia that has become an invasive species in the United States. Herbarium records suggest Florida or North Carolina are the initial site of introduction during the early 1900’s, but the fern is now established in much of the southeast including Alabama, Arkansas, Georgia, Louisiana, Mississippi, South Carolina and Texas. We aimed to ask three questions regarding the introduction of L. japonicum: (1) Was there a single Japanese Climbing Fern introduction or multiple introductions? (2) What is the distribution of haplotypes in the United States? and (3) What are the source population(s) from the native range in Asia? To answer these questions, we sequenced the chloroplast genome from 74 L. japonicum herbarium specimens representing 24 native and 50 invasive range populations. Seventeen haplotypes were found in the native range compared to three in the invasive range. Our results indicate L. japonicum has low genotype diversity in the invasive range relative to the native range. Even with low genotype diversity, this data suggests at least three introductions of L. japonicum. However, the native source population(s) of invasive L. japonicum remains unknown. These findings add to our understanding of invasive species introductions, and may have implications for biological control.