Introduced populations of organisms typically have reduced diversity compared to those that are native. It is, therefore, unusual that introduced populations of the fungal tree pathogen Diplodia pinea have been shown to have high levels of genetic diversity, even surpassing diversity in some native regions. This is thought to be due to multiple introductions over time or the existence of a cryptic and yet undiscovered sexual cycle. In this study, we consider whether populations of D. pinea in Southern Hemisphere countries have similar patterns of diversity, share some level of genetic identity and how they might be influenced by sexual recombination. A total of 173 isolates from Argentina, Australia, Ethiopia and South Africa were characterized using 12 microsatellite markers. The results show that all these populations have high gene and genotype diversities, with the Australian population having the lowest diversity. Very few private alleles were found, suggesting that isolates from different countries might share a source of introduction. However, based on allele distribution and frequency, each of the populations appeared to be evolving independently. The results showed that in all but the Australian population, alleles are randomly associated, suggesting that widespread sexual recombination has influenced population structure. ; The DST/NRF Centre of Excellence in Tree Health Biotechnology (CTHB), members of the Tree Protection Co-operative Program (TPCP) and the International Foundation for Sciences, Stockholm, Sweden, through a grant to Wubetu Bihon. ; http://link.springer.com/journal/13313 ; hb2017 ; Geography, Geoinformatics and Meteorology ; Geology