Neutron radiography (NRG) was applied to trace the development of discolored tissue in the wood of Japanese cedar (Cryptomeria japonica) after being infected with a canker fungus. Japanese cedar seedlings were wound inoculated with a virulent and avirulent isolate of a canker fungus, Guignardia cryptomeriae. Three, 7, 13 and 22 days after the inoculation, the seedlings were irradiated with thermal neutrons. The image on the X-ray film showed that the whiteness in the image corresponded to the water content in the sample. Discolored tissue and surrounding dry zones induced by the fungal inoculation were detected as dark areas, indicating water deficiency with a high resolution. Through image analysis, the dry zones were detected as early as 3 days after inoculation. Neutron images also showed the difference in the size of water deficient parts due to the tissue damage among the treatments. The neutron beam dose used in this experiment had no effect on the growth rate of the fungus on a medium, showing that NRG is an effective method for pathological research of trees.