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Evaluation of the pattern of smoking prevalence over time could provide insight for disease prevention and tobacco control policy. Examination of assortative mating is important in assessing the relative contribution of genetic and environmental influences in behavior genetics. A significant spousal concordance for smoking could imply assortative mating behavior and have potential influence on the risk for offspring smoking. Using intergenerational data from large scale adult twin panels in Australia, the present study examined the patterns of prevalence and spousal concordance of lifetime smoking across a wide range of birth cohorts. Data were collected for more than 16,000 twins, their spouses, parents, and parents-reported grandparents. The measurement models were used to calculate prevalence and spousal concordance for smoking. Prevalence of lifetime smoking for females increased significantly over successive cohorts, and was higher for twins (0.28-0.49) than for their parents or grandparents (0.06-0.42). Smoking prevalence was still higher in males than in females for the twins (0.49-0.69), and parental or grandparental (0.62-0.77) generations, but the gender differences largely decreased in younger cohorts. Moderate but significant spousal correlations were found across cohorts and generations, which were higher for twins (0.39-0.55) than for their parents and grandparents (0.19-0.49). Despite using a simplified smoking assessment, findings in this study demonstrated changes of lifetime smoking patterns across birth cohorts and the presence of assortative mating for smoking behavior.