Purpose: To establish levels of association between physical fitness and match activity profiles of players in the Australian Football League (AFL) participation pathway. Methods: Players (N = 287, range 10.9–19.1 y) were assessed on 20-m sprint, AFL agility, vertical jump and running vertical jump, 20-m multistage fitness test (MSFT), and Athletic Abilities Assessment. Match activity profiles were obtained from global positioning system measures: relative speed, maximal velocity, and relative high-speed running. Results: Correlational analyses revealed moderate relationships between sprint (r = .32–.57, P ≤ .05) and jump test scores (r = .34–.78, P ≤ .05) and match activity profiles in Local U12, Local U14, National U16, and National U18s, except jump tests in National U18s. AFL agility was also moderate to strongly associated in Local U12, Local U14, Local U18, and National U16s (r = .37–.87, P ≤ .05) and strongly associated with relative speed in Local U18s (r = .84, P ≤ .05). Match relative speed and high-speed running were moderate to strongly associated with 20-m MSFT in Local U14, Local U18, and National U18s (r = .41–.95, P ≤ .05) and Athletic Abilities Assessment in Local U12 and Local U18s (r = .35–.67, P ≤ .05). Match activity profile demands increased between Local U12 and National U16s, then plateaued. Conclusions: Physical fitness relates more strongly to match activity profiles in younger adolescent and national-level players. Recruiters should consider adolescent physical fitness and match activity profiles as dynamic across the AFL participation pathway.