New Zealand's endemic sand dune Latrodectus widow spider species, L. katipo and L. atritus, possess behavioral and physiological attributes likely to promote dispersal over large distances. Morphological, physiological and behavioral similarities between L. katipo and L. hasselti, an Australian endemic, suggest gene flow may occur across the Tasman Sea. In this study we examine intraspecific and interspecific genetic relationships within the ND1 gene region between L. katipo, L. atritus, L. hasselti and L. hesperus to assess whether the genetic evidence supports current taxonomic species designations. We found low interspecific pairwise distances among L. katipo and L. atritus populations, suggesting either introgression, incomplete lineage sorting, or that the current taxonomic distinction between the two species may be invalid. Parsimony and maximum likelihood analyses were inconclusive as to the relationships between the New Zealand Latrodectus species and the Australian L. hasselti. Low pairwise distances between L. hasselti and the New Zealand widow fauna indicated that L. katipo and L. atritus were not present in New Zealand before the fragmentation of Gondwana.