Graphitic carbon nitride (g-C3N4) microspheres (CNMS) were fabricated via a solvothermal method by using dicyandiamide and cyanuric chloride as precursors. The morphology, band structure, and defects can be simultaneously regulated by merely adjusting the concentration of precursors. Structural characterization results indicate that all the prepared samples possess spherical morphology, while the band gap decreased as the precursor concentration increased from 8 mmol (CNMS-1) to 24 mmol (CNMS-3). Besides, ultraviolet photoelectron spectroscopy results suggested that the valence band of CNMS-2 (16 mmol) was much higher than that of CNMS-1 and CNMS-3. Additionally, organic elemental analysis, X-ray photoelectron spectroscopy, and electron paramagnetic resonance results unveil the formation of nitrogen defects on the surface of prepared samples. Besides, CNMS-2 exhibits an enhanced apparent reaction rate constant of RhB degradation than that of CNMS-1 and CNMS-3. The improved apparent reaction rate constant may be due to the lowered valence band as well as the formation of nitrogen defects. This work might guide the regulation of the morphology and band structure of g-C3N4-based materials prepared via the one-pot hydrothermal method.