In order to investigate the effect of chemical and biological nitrogen fertilizer on canola, a field experiment was conducted as a split plot based on a randomized complete block design with three replications at the research station of the Faculty of Agriculture, Yasouj University, located in Deshtrum region, in 2019. The main factor consisted of nitrogen fertilizer (zero, 25, 50, 75, 100 and 125 kg/ha of pure nitrogen from urea source) and the secondary factor consisted of nitroxin biofertilizer contained Azotobacter and Azospirillium bacteria (use and non-use). The results showed that nitrogen fertilizer and nitroxin had a significant effect on all investigated traits. Nitrogen fertilizer significantly increased plant height, number of lateral branches, chlorophyll index and oil yield. The highest grain (8790.60 kg/ha) and biological (23910.93 kg/ha) yield were obtained in 125 kg/ha of nitrogen and the use of nitroxin. With the consumption of 125 kg/ha of nitrogen and nitroxin, the lowest percentage of seed oil, and contrastingly the highest percentage of seed protein (36.65%) was obtained. In general, combined application of biofertilizer with chemical nitrogen increased grain, biological and oil yield. Nitroxin bio-fertilizer could reduce the consumption of chemical nitrogen fertilizer. The results showed that there was no significant difference between the application of 50 kg/ha of nitrogen + bacteria and the application of higher levels of nitrogen (75, 100 and 125 kg/ha) alone in most of the investigated traits.