Abstract: To clarify the microstructure difference between edible fungi cultivation material and mushroom residues cellulose, in this experiment, shiitake cultivation material and shiitake mushroom residues were used as materials, the changes of microstructure of cellulose before and after shiitake cultivation were observed by scanning electrical microscopy (SEM), and the changes of cellulose crystallinity and molecular groups before and after shiitake cultivation were also studied. SEM results showed that there was little change of cellulose microstructure after edible fungi cultivation; however, small gaps appeared on cellulose surfaces. The X-ray diffraction (XRD) and FT-IR measurements showed that the crystallinity of the cellulose crystalizing region and the intensity of the cellulose characteristic absorption peak of shiitake mushroom residues were lower than those of shiitake cultivation material, which indicated that the crystallinity structure of cellulose was destroyed. This agreed with the SEM result. After shiitake cultivation, inter- and inner- hydrogen bond of cellulose structure was reset, that was the reason for the difficult degradation of cellulose.