Cotton yield and quality have been severely compromised by soil sickness throughout the primary cottongrowing regions of China. The aim of this study was to gain insight into the role of rhizosphere microbial community in governing soil sickness of cotton. Plant growth, disease resistance, root exudates, and the composition of the rhizosphere microbial community of cotton were analyzed in two different (4-and 15-year) monocropped soils and in a fallow agricultural soil (control). The monocropped soils significantly influenced cotton growth and root exudates and reduced soil suppressiveness to Fusarium wilt in bioassay experiments. Additionally, pyrosequencing of the whole internal transcribed spacers (ITS1 and ITS2) and 16S rRNA gene amplicons demonstrated clear variations in the microbial composition of cotton rhizosphere between monocropped rhizosphere soils and control soil. Specifically, monocropped soils were characterized by an increase in the abundance of fungal pathogens, including Fusarium oxysporum f. sp. vasinfectum and Verticillium dahliae, relative to the rhizosphere of control soil. Some plant-beneficial and disease-suppressive bacterial taxa, including Xanthomonadaceae, Comamonadaceae, Oxalobacteraceae, and Opitutaceae, were associated with healthy cotton. A significant correlation existed between the presence of certain amino acids (e.g., glutamic acid and alanine) and the above identified taxa, indicating that some constituents in root exudates influenced the microbial compositions of the cotton rhizosphere to manage the disease status of plant in monocropped soils. Collectively, these results suggest that pathogenic fungal build-up and a reduction in the abundance of beneficial rhizobacteria in the rhizosphere contribute to changes in soil suppressiveness to soil-borne pathogens in monocropped soils, resulting in an aggravated level of soil sickness.