Abstract
We used matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) fingerprinting to identify 95 strains of Aspergillus, Penicillium, and Talaromyces isolated from soil, marine sediment, and plants in Brazil. Samples for mass spectrometry were composed of conidia and conidiophores directly applied to the target plate and co-crystallized with the matrix solution. Cluster analyses of MS data and phylogenetic analyses based on the second largest subunit of RNA polymerase II gene sequence (RPB2) supported the identification of 42 species in Aspergillus (n = 17), Penicillium (n = 16), and Talaromyces (n = 9). In few cases, MALDI-TOF dendrograms split strains from one species into two neighboring clusters, but strains from different species were never grouped together. Three Penicillium species delimited by MALDI-TOF MS did not correspond to known species present in the large RPB2 dataset available for this group and may represent new taxa. These results expand the scope of applications of MALDI-TOF MS to the task of screening phylogenetically diverse assemblages of morphologically similar strains derived from field collections, allowing for fast discrimination of common and rare species, and detection of possible new taxa.
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Data Availability
DNA sequence data was deposited in the NCBI database, and accession codes are provided in the electronic supplementary material 1. MALDI-TOF MS data was not made publicly available since it was generated and analyzed using proprietary hardware and software.
Code availability
Not applicable.
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Acknowledgements
The authors thank Edson Luis Rezende for technical assistance. The authors would like to thank the following colleagues that provided cultures when collaborating with our working group in past projects: Cintya Neves de Souza, Jacqueline Aparecida Takahashi, Loise Araújo Costa, Maria Conceição F. Oliveira, Mirian Salgado, Patrícia Gomes Cardoso, and Vivian Gonçalves Carvalho.
Funding
This research was funded by the Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) (Proc. 563063/2010-6) and the Fundação de Amparo à Pesquisa de São Paulo (FAPESP) (Proc. 2010/52312-8). LHP received partial funding and grants provided by CNPq (Proc. 406335/2013-5 and 308291/2013-1). Scholarships were provided by Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) (GM), CNPq (DF and LMA), and Fundação de Amparo à Pesquisa do Estado de Minas Gerais (FAPEMIG) (EB).
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LMA, LHP, and ER-F conceived the research; EBC, DF, LMA, and GMM performed the researched and analyzed data; and LMA, EBC, GMM wrote the paper.
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Supplementary Table 1
Strains of Eurotiales used in this study. (PDF 237 kb)
Supplementary Figure 1.
MALDI-TOF mass spectra from selected strains of Aspergillus, Penicillium and Talaromyces in the m/z range 2000 to 16000. Each graph is composed by 24 superimposed line spectra corresponding to all replicate spectra obtained for each strain. (PDF 572 kb)
Supplementary Figure 2.
MSP dendrogram generated by multivariate cluster analysis of Main Spectra (MSPs) from 95 strains from Aspergillus, Penicillium and Talaromyces. Species identification follows the results shown in Figures 2 - 4 of the main text. (PDF 53 kb)
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Cardoso, E.B., Ferreira, D., Moreira, G.M. et al. MALDI-TOF mass spectrometry–based identification of Eurotiales from different substrates and locations in Brazil. Mycol Progress 20, 539–548 (2021). https://doi.org/10.1007/s11557-021-01691-y
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DOI: https://doi.org/10.1007/s11557-021-01691-y