AbstractBackground and Objectives: Purslane (Portulaca oleracea L.) is an annual and C4 plant that belongs to the family of Portulacaceae. The plant is drought- and salt-tolerant which contains high amounts of beneficial omega-3 fatty acids and antioxidant vitamins. Nitrogen is the key element in soil fertility and crop production. Attention to the soil quality and health has increased in recent years, especially for sustainable production of medicinal crops. So that, for production of healthy food in industrialized countries, using natural and on-farm inputs has been considered. One of the practical ways to achieve this goal is organic and biofertilizers. Given the importance of Purslane as a medicinal plant and due to the fact that there is not detailed information about the nitrogen requirement for this plant, this study was conducted to effect of organic, biological and chemical fertilizers on yield, N and P use efficiency Purslane (Portulaca oleracea L.) in Birjand, Iran Materials and Methods: In order to investigate the effect of organic, biological and chemical fertilizers on the yield and yield components of purslane seed, an experiment was conducted in a factorial randomized complete block design with three replications in Birjand University Research Farm in crop years 2015-2016 and 2017- 2018. Experimental treatments were based on a combination of four sources of nitrogen supply: cow manure, vermicompost, fertilizer (NPK) and control (without any fertilizer application), and four types of biofertilizer including: nitroxin (including azotobacter and azospirillum), mycorrhiza (Glomus intraradices), Biosulfur (including thiobacillus with sulfur) and control (without any fertilizer). The traits studied in this experiment included: biomass yield, grain yield, nitrogen and phosphorus biomass percentage, nitrogen and phosphorus biomass content, nitrogen and phosphorus uptake efficiency, seed based nitrogen and phosphorus uptake efficiency, agronomic efficiency of nitrogen and phosphorus based They were. Results: The results showed that the effect of nitrogen sources on all studied traits (except physiological grain nitrogen efficiency) was significant. The highest grain yield (1959.99 kg.ha-1) and biomass yield (9782.43 kg.ha-1) were obtained from NPK treatment with no significant difference with cow manure. Also, the most nitrogen efficiency indices were obtained from NPK treatment. However, no significant difference was observed between agronomic efficiency of grain and cow manure. The highest phosphorus content and biomass phosphorus content were obtained from NPK treatment. However, with the use of organic and chemical fertilizers, the efficiency of phosphorus use decreased. The effect of biofertilizers on most of the studied traits (except biomass P percent) was not significant. Interaction of nitrogen sources and biofertilizers was significant on most of the studied traits (except grain yield, agronomic efficiency of N and P). Conclusions: Based on the results of this experiment, considering the environmental effects and increasing the efficiency of nitrogen use for sustainable production of purslane seed in Birjand region can be used as fertilizer replacement (NPK).