Horisawa Sakae, Iwamoto Koki
Department of Environmental Systems Engineering, Faculty of Engineering, Kochi University of Technology, Kami-gun, Kochi 782-8502, Japan.
BioTech (Basel). 2022 Jul 27;11(3):30. doi: 10.3390/biotech11030030.
The accurate identification and proper typing of basidiomycetes are required in medical, sanitary maintenance, agriculture, and biotechnology fields. A diagnostic method based on information from whole-cell proteins acquired by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) was investigated to identify wood-rotting fungi, a group of filamentous fungi. In this study, mass spectra of intracellular peptides obtained from cultured mycelia of 50 strains of 10 wood-rotting fungal species were obtained multiple times and mass spectral patterns (MSPs) consisting of peaks that characterized the fungal species or strain was created to construct an in-house database. The species identification was conducted by comparing the newly obtained raw mass spectra with the MSPs in the database using the MALDI Biotyper. The results showed that the peak patterns of the mass spectra were reproducible and matched at the strain level. A cluster analysis based on the MSPs was also conducted to examine inter-and intraspecific diversity among the tested wood-rotting basidiomycetes. Most of the fungal strains examined in this study could be identified to a species level; however, the strains belonging to could only be identified to a genus level. This was due to an intraspecific variation, so the identification accuracy could be amendable with a more enhanced database.
在医学、卫生维护、农业和生物技术领域，需要准确鉴定担子菌并进行正确的分类。研究了一种基于基质辅助激光解吸/电离飞行时间质谱（MALDI-TOF MS）获取的全细胞蛋白质信息的诊断方法，以鉴定丝状真菌中的木腐真菌。在本研究中，多次获取了从10种木腐真菌的50个菌株的培养菌丝体中得到的细胞内肽段的质谱图，并创建了由表征真菌物种或菌株的峰组成的质谱模式（MSP），以构建内部数据库。使用MALDI Biotyper将新获得的原始质谱图与数据库中的MSP进行比较，从而进行物种鉴定。结果表明，质谱图的峰模式具有可重复性，并且在菌株水平上匹配。还基于MSP进行了聚类分析，以研究受试木腐担子菌的种间和种内多样性。本研究中检测的大多数真菌菌株可以鉴定到种水平；然而，属于……的菌株只能鉴定到属水平。这是由于种内变异，因此通过更完善的数据库可以提高鉴定准确性。
