The isolation of the Hawaiian archipelago has resulted in a fauna that shows high levels of endemism. I examined the role of lifestyle, as inferred from web-building versus non-web-building behavior, in dictating the rate of differentiation and species formation within a lineage of spiders in the genus Tetragnatha from the Hawaiian Islands. This genus comprises a group of morphologically, ecologically and behaviorally diverse taxa. Included in the radiation is a 'spiny-leg' clade which never builds webs and is relatively loosely associated with a specific habitat, and a large group of web-building species which are generally more tightly associated with a given substrate and habitat. Sequences of mitochondrial cytochrome oxidase DNA provided relative estimates of the age of a clade. Both linear and logarithmic models were used to estimate rates of speciation and the relative time required for speciation for each clade. The results showed that several small clades of web-building species have a greater rate of speciation as compared to the 'spiny-leg' clade. One explanation is that the web-building species may be capable of differentiation between more closely contiguous habitats, which would be consistent with the hypothesis that ecological differentiation promotes diversification and species formation. Possible alternative explanations for the results include differences in rates of molecular evolution, for example as a consequence of differences in metabolic activity.