DEVELOPMENT OF BREEDING TECHNIQUES OF BOLETACEAE USING BIOTECHNOLOGY

1)建立了牛肝菌科三种菌根真菌：乳牛肝菌、黄牛肝菌和乳牛肝菌原生质体的制备技术。我们发现，一般来说，琼脂不利于这些真菌的原生质体再生，结冷胶更好。3)我们研究了碳水化合物等各种物质的影响，4)收集了26株牛肝菌野生菌株。5)利用线粒体DNA的RFLP技术对采自日本不同地区的11株绿色乳杆菌野生菌株进行了系统发育研究，发现11株菌可分为3种类型。6)对原生质体再生菌株的形态和氧化酶活性进行了研究，发现出现了许多突变。7)对黄牛乳杆菌的细胞学进行了研究。8)我们得出了利用原生质体技术培育这3种真菌的可能性。9)我们成功地将这3种真菌分为3种类型。用灰乳杆菌和黄牛肝菌的液体培养物接种赤松无菌苗产生菌根。10)从黑牛肝菌子实体中提取了一种新的大环内酯类酚类化合物，并对其化学结构进行了测定，将其命名为地衣内酯。

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".