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Objective To examine the effects of a multi-component, theory-based, 2.5-year intervention on children’s fruit and vegetable (F&V) consumption, preferences, knowledge and body mass index (BMI). Methods Four inner city elementary schools in the Northeastern United States were randomized to an intervention (n = 149) or control group (n = 148) in 2005. F&V consumption during school lunch (measured by plate waste), preferences, and knowledge, as well as BMI were assessed five times across 3.5 years (pre-intervention, spring 2006, 2007, 2008 and 2009). Hierarchical linear modeling was used to analyze program outcomes. Results At the first post-test assessment, children in the experimental group ate 0.28 more servings/lunch of F&V relative to children in the control group and changes in F&V consumption were found in each year throughout the program. However, this effect declined steadily across time so that by the delayed one-year follow-up period there was no difference between the groups in F&V consumption. There were persistent intervention effects on children’s knowledge. There were no effects on F&V preferences and BMI throughout the study. Conclusion Although there was initial F&V behavior change, annual measurements indicated a gradual decay of behavioral effects. These data have implications for the design of school-based F&V interventions.