The purpose of this study was to examine the relationship between nutrition knowledge and dietary behavior while controlling for academic major, year, and gender among Division III student-athletes. Sufficient nutrition is essential for athletic performance, injury reduction, and overall health. However, student-athletes face unique challenges that can disrupt optimal nutrition, including poor nutrition knowledge and access to available resources. While several studies have examined the relationship between nutrition knowledge and dietary behavior, no known studies have addressed this among Division III student-athletes. This non-experimental correlational study examined dietary behavior using the Rapid Eating and Activity Assessment for Participants (REAP-S) and nutrition knowledge using the Abridged Nutrition for Sport Knowledge Questionnaire (ANSKQ). The study included 154 Division III student-athletes who completed the survey via convenience sampling. Coaches, league commissioners, athletic administrators, and athletic support staff assisted in survey distribution. Correlation between nutrition knowledge and dietary behavior and hierarchical linear regression modeling were utilized in this study. No significant relationship was found between nutrition knowledge and dietary behavior while controlling for the identified covariates. Findings offer possibilities to further the relationship between nutrition knowledge and dietary behavior of student-athletes and encourage resources and intervention programs.