Background: There were few studies exploring the associations between depressive symptoms, anxiety disorders and health-related quality of life (HRQoL) in the general population, especially in resource limited area. The aims of this study were to assess the associations between depressive symptoms, anxiety disorders, their comorbidity and HRQoL in rural area.Methods: A total of 23496 eligible participants from Henan rural cohort were included. The Patient Health Questionnaire-2 (PHQ-2) and Generalized Anxiety Disorder-2 (GAD-2) were employed to assess depressive symptoms and anxiety disorders, respectively. HRQoL was measured via European Quality of Life Five Dimension Five Level Scale (EQ-5D-5L). Tobit regression and generalized linear model were utilized to explore the associations between depressive symptoms, anxiety disorders, their comorbidity and HRQoL.Results: A total of 1320 individuals were identified as depressive symptoms with a prevalence of 5.62%, while 1198 participants were classified as anxiety disorders with a prevalence of 5.10%. After multiple adjustment, the regression coefficients and 95% confidence interval (CI) of utility index for depressive symptoms and anxiety disorders were -0.166 (-0.182, -0.149) and -0.132 (-0.149, -0.114), respectively. The regression coefficients and 95% CI of VAS score for depressive symptoms and anxiety disorders were -7.65 (-8.60, -6.70) and -5.79 (-6.78, -4.80), respectively. Additionally, the comorbidity was strongest associated with low utility index and VAS score. These findings were observed robustly in men and women.Conclusion: Depressive symptoms, anxiety disorders and their comorbidity were negatively associated with HRQoL in rural population, which needed further efforts on preventive and treatment interventions.Clinical Trial Registration: The Henan Rural Cohort Study has been registered at Chinese Clinical Trial Register (Registration number: ChiCTR-OOC-15006699). Date of registration: 06 July, 2015. http://www.chictr.org.cn/showproj.aspx?proj=11375