Abstract: The brown spot caused by Rhizoctonia solani is one of the most common diseases of creeping bentgrass (Agrostis stolonifera) lawns. Application of 2,3-butanediol (2,3-BD ) can induce plant disease response and improve disease resistance. In the present study, creeping bentgrass was inoculated with R. solani after treatment with 2,3-BD (250 μmol/L) and then the cell microstructure of leaves was observed using a paraffin sectioning method. It was found that the disease index of creeping bentgrass without 2,3-BD reached 70.45% after 15 days from inoculation. However, the disease index of creeping bentgrass treated with 2,3-BD was significantly lower, at 21.63% (P<0.05). Compared to leaves of pathogen-infected plants untreated with 2,3-BD, creeping bentgrass leaves with both pathogen inoculation and 2,3-BD treatment had a closely arranged cell structure, smaller cavities, more papillae, and increased upper and lower epidermal cell size. The size increase of upper and lower epidermal cells of 2,3-BD induced plants, compared to uninduced plants, was a factor of 1.19 and 1.26 times, respectively, and was most significant on day 7 after inoculation. The mesophyll cell size increase was the most significant on the 15th day (×1.30). The chloroplast density 1, 7, and 15 days after inoculation was, respectively, 1.25, 1.20, and 1.24 times that of uninduced plants (P<0.05). The size of vascular bundles and ducts did not change significantly at any time, however, the degree of lignification was higher and the cortical tissue became thicker in induced plants. In conclusion, 2, 3-BD can induce resistance of creeping bentgrass to brown spot disease through change in the cell structure. There was a positive correlation between the upper and lower epidermal cell size, the mesophyll cell size, and the number of chloroplasts per cell in creeping bentgrass leaves, and the disease resistance.