The Japanese golden chanterelle commonly identified as Cantharellus cibarius was sampled 3 in a broad range of forest vegetation. A total of 90 fresh and 11 herbarium specimens were 4 examined microscopically, subjected to sequencing analysis of their nuclear ribosomal RNA 5 (rDNA) and tef-1 genes, and their characteristics were compared with those of European C. 6 cibarius. Based on morphological and ecological characteristics, basidioma samples from 7 Japan were divided into four species. While specimens of Cantharellus sp. 4 from Hokkaido 8 Island were included in the European C. cibarius clade phylogenetically, the other three 9 species formed three unique clades. Among these, Cantharellus anzutake sp. nov. is sister to 10 the clade of C. cibarius and was widely sampled from the northern limit of Honshu Island to 11 the southern limit of Kumejima Island in Ryukyu Islands. Although C. anzutake was 12 morphologically similar to C. cibarius, the two species were phylogenetically distinct. Other 13 morphologically similar but genetically distinct chanterelle species from India exhibited 14 macroscopic and microscopic differences compared with C. anzutake.

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The Japanese delicacy Tricholoma matsutake has been conducted in vitro ectomycorrhizal 2 syntheses for more than 20 y. The development of its ectomycorrhizal structures varies among 3 experimental systems. Here, we examined the effects of soil-fungus interactions on the early 4 stage of in vitro T. matsutake ectomycorrhization. Axenic Pinus densiflora seedlings were 5 transplanted into autoclaved natural inorganic soil, inoculated with the cultured mycelium of T.6 matsutake, and incubated for 90 d in vitro. Both soil type and fungal strain significantly affected 7 host plant growth; host plant growth and mycorrhization levels significantly differed among soil 8 type/fungal strain combinations. Therefore, the selection of T. matsutake strains for optimal 9 mycorrhization must take into account such fungal and soil properties.

Species of fleshy yellow Cantharellus are known as chanterelles, which are among the most popular wild edible mycorrhizal mushrooms in the world. However, pure culture isolates of Cantharellus are rare. We report an efficient isolation technique of the Japanese golden chanterelle, Cantharellus anzutake, from its ectomycorrhizal root tips. Field-sampled fresh ectomycorrhizal root tips of C. anzutake on various hosts such as pines, spruce, and oaks were vortexed with 0.005% Tween 80 solution, surface sterilized with 1% calcium hypochlorite solution, rinsed with sterilized distilled water, and placed on modified Norkrans' C (MNC) agar plate medium. Most ectomycorrhizal root tips of C. anzutake produced yellowish mycelial colonies within a few months. In contrast, tissue isolation from basidiomata provided limited cultures of C. anzutake but much contamination of bacteria and molds, even on media that contained antibiotics. The established C. anzutake cultures had clamp connections on the hyphae and contained intracellular oily droplets. These cultured isolates were identified as C. anzutake by sequence analysis of the rRNA internal transcribed spacer (ITS) region and translation elongation factor EF1-alpha (tef-1) genes.