[Objective] Exploring the effects of monosodium glutamate wastewater on root characteristics and microbial diversity in rhizosphere soil of poplar, to provide theoretical basis and technical guidance for the breeding and forestation of poplar seedlings, and the popularization and application of monosodium glutamate wastewater as well.[Methods] A pot experiment including four treatments, i. e., CK(neither urea nor monosodium glutamate wastewater was applied), CF(100% of nitrogen was provided by urea), CM(100% of nitrogen was provided by monosodium glutamate wastewater) and CM+CF(50% and 50% of nitrogen were provided by monosodium glutamate wastewater and urea, respectively), was performed. The present study was conducted to determine the effects of different treatments on root construction level, root absorption area, microbial diversity index in rhizosphere soil, and the growths of ground diameter and height of poplar seedlings.[Results] The application of monosodium glutamate wastewater significantly promoted the root growth and construction of poplar seedlings, especially the promotions of the fine root weight and the ratio of fine root were obvious. In contrast to CF treatment, the CM+CF treatment significantly increased the total absorption area and active absorption area of roots, and the increments were 76.39% and 121.48%, respectively. At the same time, the maximum value of microbial diversity index was observed by the use of CM+CF, which had significant differences with other treatments. Additionally, this treatment also obviously improved the growths of ground diameter and height of poplar seedlings, indicative of 45.30%, 22.62% and 10.78% increases in ground diameter and 42.21%, 20.65% and 11.46% increases in height over the treatments of CK, CF and CM, respectively.[Conclusion] As a result, monosodium glutamate wastewater combined with inorganic fertilizer was beneficial to improve root characteristics and microbial diversity index in rhizosphere soil, and to promote growth of poplar seedlings.