[Objective]This study aimed to determine the number of microorganisms of cotton rhizosphere in a soil at various amounts of applied nitrogen fertilizer in a red date-cotton intercropping system. [Method] In the field experiment, nitrogen fer-tilizer was applied at 0, 150, 300, 450, 600, and 750 kg/hm2, respectively. The dy-namic changes of the populations of bacteria, fungi and actinomycetes at the rhizo-sphere of cotton intercropped with red date were investigated. [Result] Microbial populations significantly increased at nitrogen fertilizer of 300 and 450 kg/hm2 during bud, flowering and bol opening periods. The numbers of bacteria and actinomycetes were higher during flowering and bol opening stages than at bud stage. The num-ber of fungi slightly changed during the entire growth period. Appropriate amount of nitrogen fertilizer (300 to 600 kg/hm2) was favorable to the survival of the microor-ganisms in the soil under the intercropping system. [Conclusion] The study provides a guideline for screening and determining the optimum amount of applied nitrogen fertilizer.