DEVELOPMENT OF BREEDING TECHNIQUES OF BOLETACEAE USING BIOTECHNOLOGY

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1) We developed techniques for preparation of protoplasts of three mycorrhizal fungi of Boletaceae ; Suillus grevillei, Suillus luteus and Boletinus cavipes.2) We developed techniques for regeneration of protoplasts of the three fungi. We found that, in general, agar is not good for protoplast regeneration of these fungi, and gellan gum is better.3) We studied the effects of various substances such as carbohydrates, amino acids and related compounds on the growth of the three fungi using Ohta medium as the basal medium.4) We collected 26 wild strains of Boletaceae.5) We studied the phylogeny of 11 wild strains of Suillus grevillei collected from various region of Japan using RFLP of mitochondrial DNA and found that 11 strains could be grouped into 3 types.6) We studied morphology and oxidase activities of regenerated strains of protoplasts of Suillus luteus and found many mutations appeared.7) We studied the cytology of Suillus luteus.8) We conclude that it is possible to breed these three fungi using protoplast techniques.9) We succeeded to produce mycorrhiza by inoculating sterilly grown seedling of Pinua densiflora with liquid cultures of Suillus grevillei and Suillus luteus.10) We extracted a novel macrolide phenolic compound from fruiting bodies of Boletus ornatipes and determined the chemical structure of the compound and named it as "ornatipolide".