Background and Objective: The present study was performed to investigate the effects of chemical, animal and biological fertilizers on morphological, physiological, biochemical, functional traits, yield components of corn 500 single-cross, as well as to introduce the best fertilizer treatment. Materials and Methods: An experiment was conducted using a factorial based on the randomized complete block design with three replications in a farm located in Dasht-e-Giv in Khoosf city of South Khorasan province in late June of 2018-2019 growing season. The experimental treatments were inoculation/ non-inoculation of seeds with phosphate Barvar-2 as the main plot, and three levels of chemical fertilizer (control, N150: P25: K150 and N300: P50: K300 per hectare) and three levels of animal fertilizer (control, 10 and 20 t. ha-1) as the subplot. Furthermore, indices of plant height, shoot dry weight, row number per cob, grain number per row, 1000-grain weight, grain yield, biological yield, harvest index, seed protein, chlorophyll a, b, total chlorophyll and carotenoids were evaluated. Results: The results indicated that seed inoculation with bio-fertilizer Barvar-2 significantly increased the dry weight of aerial parts, the number of grains per row, 1000-grain weight, grain yield, biological yield, and photosynthetic pigments. Also, the application of animal fertilizer at the level of 20 t. ha-1 and chemical fertilizer at the treatment of N300: P50: K300 significantly increased all indices studied in this study than the control treatment. According to the results, it was observed that there were significant effects for interactions of bio-fertilizer × chemical fertilizer (chlorophyll b, total chlorophyll, carotenoids and biological yield), bio-fertilizer × animal fertilizer (dry weight of aerial parts, chlorophyll b, total chlorophyll and carotenoids), chemical fertilizer × animal fertilizer (dry weight of aerial parts, biological yield, chlorophyll b and carotenoids) and biofertilizer × chemical fertilizer × animal fertilizer (carotenoids). Conclusion: In general, inoculating seeds with phosphate barvar-2, the application of 20 t. ha-1 of animal fertilizer, the application of N300: P50: K300 treatment, the interaction of biological, animal and chemical fertilizers resulted in improving yield and yield components of corn 500 single-cross under agricultural conditions of the present study area.