The Mediterranean sea cucumbers including Holothuria polii has become commercially important in international trade due to the high demand of consumers from Far East countries. Sea cucumbers fisheries is a valuable income for the regional fishermen but natural stocks endangered by overfishing in recent years. Fisheries regulations and aquaculture studies are important precautions for preserving natural stocks. All these efforts are based on reproduction biology of this species which slightly differed among regions. In this study, the reproduction biology including morphological characteristics, gonadosomatic index and gonadal development stages of H.polii at the eastern coast of Aegean Sea (Izmir, Turkey) were investigated during the reproduction (spawning) period from July to October 2018. Gonads and gonad sections of 60 sea cucumbers (120.60 ± 19.56 g) have been observed by macroscopic and microscopic inspections. Three gonadal development stages have been identified by histologic observations as mature (III), spawning (IV) and post-spawning stage (V) in gonadal tubules. Results indicate that both female and male gonads are at mature and spawning stage in July and all gonads are at post-spawning stage in October. The mean gonad weight was 12.53 ± 1.33 g at the beginning of the spawning period (July) and 1.87 ± 0.58 g at the end of the spawning period (October). Gonadosomatic index decreased from 17.53 ± 0.02% (July) to 3.37 ± 0.01% (October) after spawning. The gonadosomatic index, gonad weight, and spawning were related to the seawater temperature. According to this study, the spawning period of H.polii at the eastern coasts of the Aegean Sea starts in July and completely ends in October. As a result, the data related to the reproduction biology in spawning period of H.polii would guide stock management and artificial breeding of this species under controlled conditions.