Establishment of a method for mini-metagenomic analysis of unculturable fungi and search for novel genes

2023 Fiscal Year Final Research Report

• Project/Area Number: 21K06141
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 43060:System genome science-related
• Research Institution: Kyushu University (2023); Institute of Physical and Chemical Research (2021-2022)
• Principal Investigator: Umeyama Taichi 九州大学, 医学研究院, 特任助教 (30706370)
• Project Period (FY): 2021-04-01 – 2024-03-31
• Keywords: ミニメタゲノム解析 / 蛋白質工学 / センサー / 次世代シーケンサー

Outline of Final Research Achievements

Fungi are not only rare in the microflora, but their genetic information is encoded on multiple chromosomes, making it impossible to reconstruct the entire genome from metagenomic data. For this reason, isolation culture is essential in conventional genome analysis, and the genomes of fungi that cannot be cultured cannot be analyzed. In this study, I aimed to establish a mini-metagenomic analysis method for genome analysis of fungi that cannot be cultured, by isolating and enriching fungi from the microflora, and I developed the following methods: (1) creation of probes for magnetic separation and FACS sorting and improvement of their affinity using protein engineering methods, (2) Optimization of conditions for fungal enrichment by density gradient centrifugation, and (3) Acquisition of bacteria-specific bacteriolysis enzymes and investigation of reaction conditions.

Free Research Field

ゲノム微生物学

Academic Significance and Societal Importance of the Research Achievements

近年のシーケンス技術の進歩により微生物叢から直接細菌のゲノムを完全に解読することが可能になったことで、培養不可能な細菌のゲノム解析が精力的に進められるようになった。しかしこの手法は、複数の線状ゲノムをもつ真菌に対しては適用できないため、真菌のゲノム解析はいまだに単離培養が可能な種に限定されている。本研究を基に難培養真菌のゲノム解析が可能になれば、新規の真菌種や遺伝子の発見につながるため学術的に大きな意義がある。また真菌は宿主の免疫や神経発達に影響するため、共生微生物叢の生体防御や認知機能への影響がより詳細に理解できるようになれば、社会的に大きな意義がある。