The role of cortical resin canals in the early development of a pine wilt disease in Japanese black pine, Pinus thunbergii, was studied. A part of the bark of a 2 cm long segment from a current-year stem was removed by a tangential cut with a razor blade. Both cortex-exposed segments with cut cortical resin canals (designated as +RC segments) and those without cut resin canals (–RC segments) were obtained by chance. When a virulent nematode isolate (S6-1) was inoculated onto the cut surface, the surface of the +RC segments turned brown 4 d after inoculation, and in some segments this browning occurred more intensely around cortical resin canals. When segments were cut transversely at the middle, the transverse cut surface of the inoculated +RC segments was brown and fragile, but that of the inoculated –RC segments was pale green and stable, as was that of the non-inoculated controls. Correspondingly, tissue cells including epithelial cells of the cortical resin canal of the +RC segments were all dead, but those of the –RC segments and the controls were alive. When nematodes were inoculated onto the inner surface (cambium side) of a bark peeling at which cambial xylem cells were exposed, they did not kill the cambial cells. When inoculated on the transverse cut end of a bark peeling from which nematodes would enter resin canals, nematodes killed all cells in the peeling. The above results indicate that nematodes do not kill cortical cells immediately after inoculation but become harmful to pine cells after living in cortical resin canals. Key words: Bursaphelenchus xylophilus, pine wilt disease, pine wood nematode, Pinus thunbergii, resin canal.
The role of bacteria in pine wilt disease: insights from microbiome analysis
