Genome sequencing of evolved aspergilli populations reveals robust genomes, transversions in A. flavus, and sexual aberrancy in non-homologous end-joining mutants
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Published:2019-11-11
Issue:1
Volume:17
Page:
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ISSN:1741-7007
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Container-title:BMC Biology
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language:en
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Short-container-title:BMC Biol
Author:
Álvarez-Escribano Isidro, Sasse Christoph, Bok Jin Woo, Na Hyunsoo, Amirebrahimi Mojgan, Lipzen Anna, Schackwitz Wendy, Martin Joel, Barry Kerrie, Gutiérrez Gabriel, Cea-Sánchez Sara, Marcos Ana T., Grigoriev Igor V., Keller Nancy P., Braus Gerhard H., Cánovas DavidORCID
Abstract
AbstractBackgroundAspergillusspp. comprises a very diverse group of lower eukaryotes with a high relevance for industrial applications and clinical implications. These multinucleate species are often cultured for many generations in the laboratory, which can unknowingly propagate hidden genetic mutations. To assess the likelihood of such events, we studied the genome stability of aspergilli by using a combination of mutation accumulation (MA) lines and whole genome sequencing.ResultsWe sequenced the whole genomes of 30 asexual and 10 sexual MA lines of threeAspergillusspecies (A. flavus,A. fumigatusandA. nidulans) and estimated that each MA line accumulated mutations for over 4000 mitoses during asexual cycles. We estimated mutation rates of 4.2 × 10−11(A. flavus), 1.1 × 10−11(A. fumigatus) and 4.1 × 10−11(A. nidulans) per site per mitosis, suggesting that the genomes are very robust. Unexpectedly, we found a very high rate of GC → TA transversions only inA. flavus. In parallel, 30 asexual lines of the non-homologous end-joining (NHEJ) mutants of the three species were also allowed to accumulate mutations for the same number of mitoses. Sequencing of these NHEJ MA lines gave an estimated mutation rate of 5.1 × 10−11(A. flavus), 2.2 × 10−11(A. fumigatus) and 4.5 × 10−11(A. nidulans) per base per mitosis, which is slightly higher than in the wild-type strains and some ~ 5–6 times lower than in the yeasts. Additionally, inA. nidulans, we found a NHEJ-dependent interference of the sexual cycle that is independent of the accumulation of mutations.ConclusionsWe present for the first time direct counts of the mutation rate of filamentous fungal species and find thatAspergillusgenomes are very robust. Deletion of the NHEJ machinery results in a slight increase in the mutation rate, but at a rate we suggest is still safe to use for biotechnology purposes. Unexpectedly, we found GC→TA transversions predominated only in the speciesA. flavus, which could be generated by the hepatocarcinogen secondary metabolite aflatoxin. Lastly, a strong effect of the NHEJ mutation in self-crossing was observed and an increase in the mutations of the asexual lines was quantified.
Funder
Office of Science National Institutes of Health Deutsche Forschungsgemeinschaft Ministerio de Economía y Competitividad
Publisher
Springer Science and Business Media LLC
Subject
Cell Biology,Developmental Biology,Plant Science,General Agricultural and Biological Sciences,General Biochemistry, Genetics and Molecular Biology,Physiology,Ecology, Evolution, Behavior and Systematics,Structural Biology,Biotechnology
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