OBJECTIVE: This study examines the relationship between diet quality and health-related quality of life (HRQoL) in rural and urban Australian adolescents, and gender differences. DESIGN: Cross-sectional. SETTING: Secondary schools. PARTICIPANTS: 722 rural and 422 urban students from 19 secondary schools. MAIN OUTCOME MEASURES: Self-report dietary-related behaviours, demographic information, HRQoL (AQoL-6D) were collected. Healthy and unhealthy diet quality scores were calculated; multiple linear regression investigated associations between diet quality and HRQoL. RESULTS: Compared to urban students, rural students had higher HRQoL, higher healthy diet score, lower unhealthy diet score, consumed less soft drink and less frequently, less takeaway and a higher proportion consumed breakfast (P < 0.05). Overall, males had higher unhealthy diet score, poorer dietary behaviours but a higher HRQoL score compared to females (P < 0.05). In all students, final regression models indicated: a unit increase in healthy diet score was associated with an increase in HRQoL (unstandardised coefficient(B)±standard error(SE); B = 0.02 ± 0.01(SE); P < 0.02); and a unit increase in unhealthy diet scores was associated with a decrease in HRQoL (-0.01 ± 0.00; P < 0.05). In rural students alone, a unit increase in unhealthy diet score was associated with a decrease in HRQoL (B = -0.01 ± 0.00; P = 0.002), and in urban students a unit increase in healthy diet score was associated with an increase in HRQoL (B = 0.02 ± 0.00; P < 0.001). CONCLUSIONS: Cross-sectional associations between diet quality and HRQoL were observed. Dietary modification may offer a target to improve HRQoL and general well-being; and consequently the prevention and treatment of adolescent health problems. Such interventions should consider gender and locality.