This study was conducted to understand how osmotic potentials in Lentinula edodes tissues are related to water contents and how they change while a mushroom matures. Water content and osmotic potential of L. edodes mushroom tissues from log cultivation and sawdust cultivation were measured and the relationships were analyzed. Osmotic potentials in the tissues were exponentially proportional to their moisture contents and there were strain differences in the potentials. Strain 290 has lower osmotic potential than strain 302, in the tissues at the same water content. As the mushrooms mature, tissue water content maintained ca 94% in head tissues and ca 90% in gills, but significantly decreased from ca 90% to 82% in the stipe tissues. Osmotic potential changes were similar to the tissue water content changes as the mushrooms mature. While osmotic potentials maintained -0.25 to -0.45 MPa in head and gill tissues, the potentials greatly decreased from -0.65 to -1.33 MPa in stipe tissues. Our results show that osmotic potentials in L. edodes tissues are exponentially proportional to tissue water contents, that strains differ in osmotic potential related to water, and that stipe tissues can still have nutritional value when they mature.