Two-dimensional (2D) transition metal dichalcogenides (TMDs) exhibit enormous potential in the field of optoelectronics. The high performance of TMD materials and optoelectronic devices significantly depends on processes involved in photoelectric conversion, including photo-excitation, relaxation, transportation, and recombination. Remarkably, inevitable defects in materials prolong or shorten the characteristic time of these processes and even bring about new photoelectric conversion channels, namely, the defect-related relaxation pathways of photoexcited carriers tailor the performance of photoelectric applications. In recent years, there have been numerous investigations in exploring the variant transient signals caused by defects in TMDs utilizing ultrafast spectroscopies. They have the capability in providing an accurate and overall representation of ultrafast processes owing to the subtle temporal resolution. The defect-related mechanisms occurring in different time scales (from femtosecond (fs) to microsecond (μs)) play influential roles throughout the relaxation process of photoexcited species. Herein, we review the defect-related relaxation mechanisms of photoexcited species in TMDs according to the time scale utilizing ultrafast spectroscopy techniques. By interpreting and summarizing the defect-related transient signals, we furnish the direction in material design and performance optimization.