Tissues from Lentinula edodes were collected from various regions of the sporocarp, specifically the stipe, pileus, and the junction of the pileus and stipe, and subsequently cultured on Potato Dextrose Agar (PDA) medium. The primary objective of this study was to evaluate key yield attributes that influence mushroom cultivation. We assessed the spawn run duration, time taken for primordial initiation, average weight of sporocarps, total number of sporocarps produced, overall yield, and biological efficiency of each culture. Our findings indicate significant variations in growth characteristics among the different tissue sources. Notably, cultures derived from the junction of the pileus and stipe demonstrated superior performance, exhibiting the shortest spawn run period and the highest yield and biological efficiency. These results suggest that the anatomical location of the tissue plays a crucial role in the growth and productivity of Lentinula edodes. The insights gained from this study contribute to a better understanding of the optimal tissue selection for enhanced mushroom cultivation under controlled conditions, thereby promoting efficient production practices in the industry. Future research should further explore the physiological mechanisms that underpin these differences in yield attributes.