Considering that monoculture yeild has increased due to excessive use of chemical fertilizers and this process ultimately causes environmental pollution, therefore, in order to replace chemical fertilizers with biological fertilizers in Mixed cropping, an experiment was conducted in the agricultural year of 2018-2019 in order to evaluate the mixed cropping of wheat and canola under the influence of biological and chemical fertilizers in the research farm of the Faculty of Agriculture of Lorestan University. The experiment was performed as a split plot in the form of a randomized complete block design in three replications. The first factor was 200 kg/ha of nitrogen + 150 kg/ha of phosphorus, 100 kg/ha of nitrogen + 75 kg/ha of phosphorus + 200 kg/ha of nitrogen + 150 kg/ha of phosphorus, and the control treatment. The second factor was designed in five different planting pattern , including single crop of wheat, single crop of rapeseed, 25% wheat + 75% rapeseed, 50% wheat + 50% rapeseed and 25% rapeseed + 75% wheat. The interaction of treatments showed that different patterns of planting and fertilizer were significant on seed yield and physiological traits. The highest chlorophyll, net photosynthesis, transpiration and stomatal conductance of canola and wheat was obtained in the model of pure cultivation + combination of chemical and biological fertilizers. On the other hand, the highest stomatal resistance of canola from the lowest mixture component (25%) was observed in the presence of chemical fertilizer. It is worth mentioning that the highest stomatal resistance of wheat was obtained from 25% of wheat in the presence of combination of biological and chemical fertilizers. Considering that the highest yield and other traits were observed from pure cultivation, but the total results showed the necessary superiority in treatments of 75:25 and 50:50 in rapeseed and wheat compared to pure cultivation. According to the present experiment, the use of combined fertilizers (chemical + biological fertilizers) improved the above traits, so it is recommended to grow a mixture of these two crops in the ratio of 75% wheat and 25% rapeseed instead of a single crop of wheat and rapeseed. The use of pure chemical fertilizers, biological fertilizers should be used in combination with chemical fertilizers.