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To evaluate the response of saffron to animal manure, and biological and chemical fertilizer in an arid climate, an experiment was performed as split plots based on a randomized complete blocks design with three replications during three consecutive crop growth seasons (2015–2018) at the Research Farm of University of Gonabad, Iran. The experimental treatments included application (60 t ha−1) and non-application (control) of manure as the main plot and the use of biosulfur (5 kg ha−1), biophosphate (3 L ha−1), nitroxin (3 L ha−1), chemical fertilizer (150, 100, and 100 kg ha−1 of urea, triple superphosphate, and potassium sulfate, respectively), and no fertilizer application (control) as the sub-plot. The results showed a highly significant response of the quantitative traits of saffron to the application of manure, which increased the leaf, flower, and corm indices of saffron by a mean of 15.1–35.7% than control. The interaction effect of manure with biological and chemical fertilizers for leaf, flower, and weeds indices of saffron was significant. There was no significant difference between the interaction treatments of manure and chemical fertilizer with nitroxin and biophosphorus fertilizers in most of the mentioned traits in the three experiment years. The simultaneous application of these fertilizers increased the average by about 60, 105, 135, 110, 165, and 55% of the leaf dry weight, the number of flowers, fresh flower yield, dry flower yield, dry stigma yield, and weed dry weight of saffron, respectively as compared to control. There was no significant difference between the chemical fertilizer with nitroxin or biophosphate in terms of the effect on the traits related to saffron corm so the use of these fertilizers, as compared to control, increased replacement corm weight, replacement corm size, and bud number per corm by, respectively, about 35, 60, and 40% on average. The chemical and biological fertilizers improved the content of crocin, picrocrocin, and safranal of saffron stigma. The best results were obtained from the use of chemical fertilizers, although no significant difference was observed between this treatment and the nitroxin and biophosphate treatments. Overall, the results of this three-year experiment show a very high response of the saffron plant to the simultaneous use of manure and biological fertilizers and, therefore, it is possible to replace chemical fertilizers with organic and biological fertilizers in saffron cultivation to implement organic agriculture and achieve acceptable quantitative and qualitative yields in areas similar to the experiment location. [Display omitted] • The effect of manure on the saffron traits was significantly positive. • The combination of manure and biological or chemical fertilizers improved saffron crop. • The combination of manure and chemical fertilizer or biophosphate improved saffron stigma quality. • Biophosphate and nitroxin biofertilizer improved saffron yield. [ABSTRACT FROM AUTHOR]