Studies on transformation methods of Tricholoma matsutake, and isolation of monokaryon of the fungus

2021 Fiscal Year Final Research Report

• Project/Area Number: 19K06175
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 40020:Wood science-related
• Research Institution: Kindai University
• Principal Investigator: Fukuta Yasuhisa 近畿大学, 農学部, 准教授 (80609602)
• Project Period (FY): 2019-04-01 – 2022-03-31
• Keywords: マツタケ / トキイロヒラタケ / 形質転換 / 単核体 / 担子菌 / 細胞壁 / プロトプラスト

Outline of Final Research Achievements

With the aim of establishing a transformation method for Tricholoma matsutake, research was conducted with the aim of isolation of monokaryon as a host strain, and developing genetic recombination technology using the protoplast-PEG method in the fungi. However, protoplasts could not be regenerated during the study period. Currently, we are using α-1,3-glucanase derived from the cell wall of the fungi component degradation microorganism isolated from nature to adjust protoplasts in an even larger amount, and continue regeneration experiments. On the other hand, as a genetic recombination experiment using the same basidiomycetes, Pleurotus salmoneostramineus, which can form fruiting bodies, was used in a model experiment. By the method established in this experiment, the expression of the pink colored protein gene (PsPCP) was suppressed, and white fruiting bodies were formed in the experiment.

Free Research Field

応用微生物学

Academic Significance and Societal Importance of the Research Achievements

マツタケのプロトプラストを再生させることができていないため、形質転換には成功していない。しかしながら、マツタケ細胞壁分解微生物が生産する酵素群を利用してプロトプラスト量を増加させることで再生率を上げることが期待できる。また新たな知見としてマツタケ細胞壁消化にα-1,3-グルカナーゼが重要な役割があることが示唆された。一方、トキイロヒラタケの形質転換法は確立され、塩基配列が明らかであれば、特定遺伝子の発現抑制がある程度可能となった。本研究で開発した手法により、子実体形成に関わる遺伝子を網羅的に抑制させて表現型を観察することで、子実体形成に必須の遺伝子を特定することができる。