Abstract: Background and Objective: Quinoa (Chenopodium quinoa Willd.), a pseudocereal is a new introduction in Thailand. The production of high grain quality and quantity which depending on various growth factor. This study focusing on planting location, nitrogen management and cultivars. This experiment was aimed to evaluate the physiological characteristics, yield and seed quality of quinoa responses to different nitrogen fertilizer rates in Thailand climate condition. Methodology: The experimental design was split-split plot in Randomized Complete Block Design (RCBD) with four replications. Main plot was planting locations (Pang-Da Research Station, Chiang Mai, Thailand (PD) and Prabathuaytom Royal Project Development Center Plant, Lamphun, Thailand (PB). Sub-plot was nitrogen fertilizer rate (0, 93.75, 187.5 and 312.5 kg N ha–1) and sub-sub plot was quinoa cultivar (Moradas and Verdes). Results: The study found that at PD location showed higher of Leaf Area Index (LAI), Leaf Area Ratio (LAR), seed yield and 1,000 seed weight (1.47, 31.97 cm2 g–1, 2144.20 kg ha–1 and 3.00 g, respectively) over than PB location. On the other hand, PB location showed higher of seed germination (97%), germination index (7.86) and low of mean germination time (3.12 days) when compared with PD location. Furthermore, nitrogen fertilizer rate at 93.75, 187.5 and 312.5 kg N ha–1 showed highest of Leaf Area Index (LAI) (1.04, 1.21 and 1.14, respectively) and Crop Growth Rate (CGR) (1.87, 1.85 and 1.48 mg cm–2 day–1, respectively). Then, nitrogen fertilizer rate at 93.75 and 187.5 kg N ha–1 showed highest of Relative Growth Rate (RGR) (35.54 and 32.59 mg g–1 day–1, respectively). Additionally, nitrogen fertilizer rate at 93.75, 187.5 and 312.5 kg N ha–1 resulted higher of seed yield (1872.00, 1726.70 and 1822.20 kg ha–1, respectively) than those control. Nitrogen fertilizer rate at 187.5 kg N ha–1 revealed higher of 1,000 seed weight at (3.02 g) than 93.75 kg N ha–1 and control. Therefore, Verdes cultivar provided the good physiological responses than those Moradas cultivar. Verdes cultivar showed higher of CGR ( 1.87 mg cm–2 day–1), RGR (31.80 mg g–1 day–1), seed yield (1892.80 kg ha–1) and 1,000 seed weight (2.97 g) over than Moradas cultivar. Conclusion: Finally, it could be concluded that at PD research station was high responded to that nitrogen fertilizer rates and cultivars for increased crop physiological growth and improved seed yield of quinoa production in Thailand.