Objective: To investigate whether practice rurality and rural training pathway are associated with general practitioner registrars' participation in their practice's after-hours care roster. Design: A cross-sectional analysis of data (2017-2019) from the Registrar Clinical Encounters in Training study, an ongoing inception cohort study of Australian general practitioner registrars. The principal analyses used logistic regression. Setting: Three national general practitioner regional training organisations across 3 Australian states. Participants: General practitioner registrars in training within regional training organisations. Main outcome measure: Involvement in practice after-hours care was indicated by a dichotomous response on a 6-monthly Registrar Clinical Encounters in Training study questionnaire item. Results: 1576 registrars provided 3158 observations (response rate 90.3%). Of these, 1574 (48.6% [95% confidence interval: 46.8-50.3]) involved registrars contributing to their practice's after-hours roster. In major cities, 40% of registrar terms involved contribution to their practice's after-hours roster; in regional and remote practices, 62% contributed to the after-hours roster. On multivariable analysis, both level of rurality of practice (odds ratio(OR) 1.75, P = .007; and OR 1.74, P = .026 for inner regional and outer regional/remote locations, respectively, versus major city) and rural training pathway of registrar (OR 1.65, P = .008) were significantly associated with more after-hours roster contribution. Other associations were registrars' later training stage, larger practices and practices not routinely bulk billing. Significant regional variability in after-hours care was identified (after adjusting for rurality). Conclusion: These findings suggest that registrars working rurally and those training on the rural pathway are more often participating in practice after-hours rosters. This has workforce implications, and implications for the educational richness of registrars' training environment.