Gardening has been positively associated with fruit and vegetable (FV) consumption based on short-term studies among children, but long-term data among adolescents and young adults are lacking. This investigation sought to elucidate the association between gardening experience and FV intake among college students over a two-year period. Students (N = 593) from eight universities were assessed at the end of their freshman (Y1) and sophomore (Y2) years during the springs of 2016 and 2017, respectively. At each time point, participants completed the NCI FV Screener and questions related to gardening experience and FV-related attitudes and behaviors. Students were then categorized into four groups based on gardening experience: Gardened only during the first or second year (Y1 only and Y2 only gardeners), gardened both years (Y1+Y2 gardeners), and non-gardeners. While both Y1 only and Y1+Y2 gardeners reported significantly higher FV intake relative to non-gardeners at Y1 (2.3 ± 0.9 and 2.6 ± 0.7 versus 1.9 ± 0.6 cup equivalents (CE)/day, respectively; p < 0.01), only Y1+Y2 gardeners differed from non-gardeners at Y2 (2.4 ± 0.6 versus 1.8 ± 0.5 CE/day; p < 0.001). Additionally, Y1+Y2 gardeners reported more frequent engagement of several FV-related behaviors, including shopping at farmers’ markets, eating locally grown foods, and cooking from basic ingredients; and were five times more likely to have gardened during childhood (OR: 5.2, 95%, CI: 3.5–8.8; p < 0.001). Findings suggest that while isolated gardening experiences during college are associated with FV intake, reoccurring experience may be essential for sustained benefit.