Global population genomics redefines domestication and clinical diversity in the Aspergillus flavus–oryzae complex
Authors/Creators
- 1. University of Debrecen, Debrecen, Hungary
- 2. North Carolina State University, Raleigh, United States of America
- 3. University of Debrecen, Debrecen, Hungary|Food Chain Safety Laboratory Directorate, Budapest, Hungary
Description
Aspergillus flavus is a globally important human pathogen and agricultural contaminant, while its domesticated relative A. oryzae is widely used in food fermentation and biotechnology. Despite their importance, the evolutionary relationship, population structure and domestication history of these fungi remain unresolved. Here, we present the first global population genomic analysis of 639 A. flavus and A. oryzae isolates from clinical, environmental and food-fermentation sources across multiple continents. Our analyses reveal a complex evolutionary landscape comprising well-separated clades interspersed with highly admixed mosaic groups and potential evidence for multiple independent domestication events giving rise to A. oryzae. Clinical A. flavus isolates are distributed across several clades and mosaic groups, some overlapping with fermentation strains, highlighting an apparent role of domestication and admixture in shaping pathogen diversity. These results challenge current species boundaries and provide a framework for understanding evolutionary history, taxonomy and pangenomic architecture in these fungi, with broad implications for pathogenicity, food safety, biocontrol and metagenomic surveillance.
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References
- Ajmal M, Alshannaq AF, Moon H et al. (2022) Characterization of 260 isolates of Aspergillus section Flavi obtained from sesame seeds in Punjab, Pakistan. Toxins (Basel) 14: 117. https://doi.org/10.3390/TOXINS14020117
- Alshannaq AF, Gibbons JG, Lee MK et al. (2018) Controlling aflatoxin contamination and propagation of Aspergillus flavus by a soy-fermenting Aspergillus oryzae strain. Scientific Reports 8: 1–14. https://doi.org/10.1038/s41598-018-35246-1
- Alvarez F, Arena M, Auteri D et al. (2022) Peer review of the pesticide risk assessment of the active substance Aspergillus flavus strain MUCL54911. EFSA journal. European Food Safety Authority 20: e07202. https://doi.org/10.2903/J.EFSA.2022.7202
- Arias RS, Orner VA, Martinez-Castillo J et al. (2021) Aspergillus section Flavi, need for a robust taxonomy. Microbiology Resource Announcements 10: 99–101. https://doi.org/10.1128/mra.00784-21
- Arias RS, Mohammed A, Orner VA et al. (2020) Sixteen draft genome sequences representing the genetic diversity of Aspergillus flavus and Aspergillus parasiticus colonizing peanut seeds in Ethiopia. Microbiology Resource Announcements 9: e00591-20. https://doi.org/10.1128/MRA.00591-20
- Augusto J, Atehnkeng J, Ortega-Beltran A et al. (2024) Keeping toxigenic Aspergillus section Flavi and aflatoxin contamination at bay by deploying atoxigenic-based biocontrol products during production of groundnut and maize in Mozambique. Frontiers in Microbiology 15: 1501924. https://doi.org/10.3389/FMICB.2024.1501924
- Van der Auwera GA, Carneiro MO, Hartl C et al. (2013) From fastQ data to high-confidence variant calls: The genome analysis toolkit best practices pipeline. Current Protocols in Bioinformatics 43: 11.10.1-11.10.33. https://doi.org/10.1002/0471250953.bi1110s43
- Bal J, Yun SH, Chun J et al. (2016) Taxonomic characterization, evaluation of toxigenicity, and saccharification capability of Aspergillus section Flavi isolates from Korean traditional wheat-based fermentation starter nuruk. Mycobiology 44: 155–161. https://doi.org/10.5941/MYCO.2016.44.3.155
- Basenko EY, Shanmugasundram A, Böhme U et al. (2024) What is new in FungiDB: a web-based bioinformatics platform for omics-scale data analysis for fungal and oomycete species. Genetics 227: iyae035. https://doi.org/10.1093/GENETICS/IYAE035
- Buil JB, Houbraken J, Reijers MH et al. (2021) Genetic and phenotypic characterization of in-host developed azole-resistant Aspergillus flavus isolates. Journal of Fungi (Basel) 7: 164. https://doi.org/10.3390/JOF7030164
- Carbone I, Ramirez-Prado JH, Jakobek JL et al. (2007) Gene duplication, modularity and adaptation in the evolution of the aflatoxin gene cluster. BMC Evolutionary Biology 7: 111. https://doi.org/10.1186/1471-2148-7-111
- Chacón-Vargas K, McCarthy CO, Choi D et al. (2021) Comparison of two Aspergillus oryzae genomes from different clades reveals independent evolution of alpha-amylase duplication, variation in secondary metabolism genes, and differences in primary metabolism. Frontiers in Microbiology 12: 691296. https://doi.org/10.3389/FMICB.2021.691296
- Chang CC, Chow CC, Tellier LCAM et al. (2015) Second-generation PLINK: rising to the challenge of larger and richer datasets. GigaScience 4: 7. https://doi.org/10.1186/S13742-015-0047-8
- Chang PK, Ehrlich KC, Hua SST (2006) Cladal relatedness among Aspergillus oryzae isolates and Aspergillus flavus S and L morphotype isolates. International Journal of Food Microbiology 108: 172–177. https://doi.org/10.1016/J.IJFOODMICRO.2005.11.008
- Chen S, Zhou Y, Chen Y et al. (2018) fastp: an ultra-fast all-in-one FASTQ preprocessor. Bioinformatics 34: i884–i890. https://doi.org/10.1093/bioinformatics/bty560
- Chin YW, Hong SP, Lim SD et al. (2024) Investigation of microbial community of Korean soy sauce (ganjang) using shotgun metagenomic sequencing and its relationship with sensory characteristics. Microorganisms 12: 2559. https://doi.org/10.3390/microorganisms12122559
- Choi D, Alshannaq AF, Bok Y et al. (2024) Broad-spectrum antimicrobial activities of a food fermentate of Aspergillus oryzae. Microbiology Spectrum 12: e0185424. https://doi.org/10.1128/spectrum.01854-24
- Cohn F (1884) Über Schimmelpilze aIs Gährungserreger. Jahresbericht der Schlesischen Gesellschaft für Vaterländische Kultur 61: 226–229.
- Costantin M, Lucet M (1905) Quelques Aspergillus pathogènes. Annales des Sciences Naturelles. Botanique 9: 119–171.
- Dabas Y, Xess I, Pandey M et al. (2022) Epidemiology and antifungal susceptibility patterns of invasive fungal infections (IFIs) in India: A prospective observational study. Journal of Fungi (Basel) 8: 33. https://doi.org/10.3390/JOF8010033
- Dorner JW (2004) Biological control of aflatoxin contamination of crops. Journal of Toxicology: Toxin Reviews 23: 425–450. https://doi.org/10.1081/TXR-200027877
- Drott MT, Rush TA, Satterlee TR et al. (2021) Microevolution in the pansecondary metabolome of Aspergillus flavus and its potential macroevolutionary implications for filamentous fungi. Proceedings of the National Academy of Sciences of the United States of America 118: e2021683118. https://doi.org/10.1073/pnas.202168311
- Drott MT, Wang Y-W, Hatmaker EA et al. (2025) Population-specific transcriptomic shifts underlie secondary metabolic diversification in Aspergillus flavus and the domestication of Aspergillus oryzae. bioRxiv 2025.10.02.680074. https://doi.org/10.1101/2025.10.02.680074
- Drott MT, Satterlee TR, Skerker JM et al. (2020) The frequency of sex: Population genomics reveals differences in recombination and population structure of the aflatoxin-producing fungus Aspergillus flavus. mBio 11: 1–13. https://doi.org/10.1128/MBIO.00963-20
- Fountain JC, Clevenger JP, Nadon B et al. (2020) Draft genome sequences of one Aspergillus parasiticus isolate and nine Aspergillus flavus isolates with varying stress tolerance and aflatoxin production. Microbiology Resource Announcements 9: e00478-20. https://doi.org/10.1128/MRA.00478-20
- Frisvad JC, Hubka V, Ezekiel CN et al. (2019) Taxonomy of Aspergillus section Flavi and their production of aflatoxins, ochratoxins and other mycotoxins. Studies in Mycology 93: 1–63. https://doi.org/10.1016/j.simyco.2018.06.001
- Galili T, O'Callaghan A, Sidi J et al. (2018) heatmaply: an R package for creating interactive cluster heatmaps for online publishing. Bioinformatics 34: 1600–1602. https://doi.org/10.1093/BIOINFORMATICS/BTX657
- Gangurde SS, Korani W, Bajaj P et al. (2024) Aspergillus flavus pangenome (AflaPan) uncovers novel aflatoxin and secondary metabolite associated gene clusters. BMC Plant Biology 24: 1–17. https://doi.org/10.1186/s12870-024-04950-8
- Geiser DM, Pitt JI, Taylor JW (1998) Cryptic speciation and recombination in the aflatoxin-producing fungus Aspergillus flavus. Proceedings of the National Academy of Sciences of the United States of America 95: 388–393. https://doi.org/10.1073/PNAS.95.1.388
- Geiser DM, Dorner JW, Horn BW et al. (2000) The phylogenetics of mycotoxin and sclerotium production in Aspergillus flavus and Aspergillus oryzae. Fungal Genetics and Biology 31: 169–179. https://doi.org/10.1006/FGBI.2000.1215
- Gell RM, Horn BW, Carbone I (2020) Genetic map and heritability of Aspergillus flavus. Fungal Genetics and Biology 144: 103478. https://doi.org/10.1016/J.FGB.2020.103478
- Gibbons JG, Salichos L, Slot JC et al. (2012) The evolutionary imprint of domestication on genome variation and function of the filamentous fungus Aspergillus oryzae. Current Biology 22: 1403–1409. https://doi.org/10.1016/J.CUB.2012.05.033
- Glöckner FO, Yilmaz P, Quast C et al. (2017) 25 years of serving the community with ribosomal RNA gene reference databases and tools. Journal of Biotechnology 261: 169–176. https://doi.org/10.1016/J.JBIOTEC.2017.06.1198
- Han DM, Baek JH, Choi DG et al. (2024) Comparative pangenome analysis of Aspergillus flavus and Aspergillus oryzae reveals their phylogenetic, genomic, and metabolic homogeneity. Food Microbiology 119: 104435. https://doi.org/10.1016/J.FM.2023.104435
- Hatmaker EA, Barber AE, Drott MT et al. (2025) Population structure in a fungal human pathogen is potentially linked to pathogenicity. Nature Communications 16: 1–12. https://doi.org/10.1038/s41467-025-62777-9
- Hatmaker EA, Barber AE, Drott MT et al. (2024) Pathogenicity is associated with population structure in a fungal pathogen of humans. bioRxiv 2024.07.05.602241 https://doi.org/10.1101/2024.07.05.602241
- Hill JH, Round JL (2024) Intestinal fungal-host interactions in promoting and maintaining health. Cell Host & Microbe 32: 1668–1680. https://doi.org/10.1016/J.CHOM.2024.09.010
- Horn BW (2003) Ecology and population biology of aflatoxigenic fungi in soil. Journal of Toxicology: Toxin Reviews 22: 351–379. https://doi.org/10.1081/TXR-120024098
- Houbraken J, Kocsubé S, Visagie CM et al. (2020) Classification of Aspergillus, Penicillium, Talaromyces and related genera (Eurotiales): An overview of families, genera, subgenera, sections, series and species. Studies in Mycology 95: 5–169. https://doi.org/10.1016/J.SIMYCO.2020.05.002
- Houbraken J, de Vries RP, Samson RA (2014) Modern taxonomy of biotechnologically important Aspergillus and Penicillium species. Advances in Applied Microbiology 86: 199–249. https://doi.org/10.1016/B978-0-12-800262-9.00004-4
- Houbraken J, Visagie CM, Frisvad JC (2021) Recommendations to prevent taxonomic misidentification of genome-sequenced fungal strains. Microbiology Resource Announcements 10: e0107420. https://doi.org/10.1128/MRA.01074-20
- Huson DH, Bryant D (2006) Application of phylogenetic networks in evolutionary studies. Molecular Biology and Evolution 23: 254–267. https://doi.org/10.1093/MOLBEV/MSJ030
- Hyde KD, Al-Hatmi AMS, Andersen B et al. (2018) The world's ten most feared fungi. Fungal Diversity 93: 161–194. https://doi.org/10.1007/s13225-018-0413-9
- Jeong E, Seo JA (2025) Comparative genome analyses of Aspergillus oryzae and Aspergillus flavus originated from a Korean fermentation starter, nuruk. Food Microbiology 131. https://doi.org/10.1016/j.fm.2025.104807
- Kalyaanamoorthy S, Minh BQ, Wong TKF et al. (2017) ModelFinder: Fast model selection for accurate phylogenetic estimates. Nature Methods 14: 587. https://doi.org/10.1038/NMETH.4285
- Kanaujia R, Singh S, Rudramurthy SM (2023) Aspergillosis: an update on clinical spectrum, diagnostic schemes, and management. Current Fungal Infection Reports 17: 144–155. https://doi.org/10.1007/S12281-023-00461-5
- Khan AW, Garg V, Roorkiwal M et al. (2020) Super-pangenome by integrating the wild side of a species for accelerated crop improvement. Trends in Plant Science 25: 148–158. https://doi.org/10.1016/j.tplants.2019.10.012
- Khan R, Ghazali FM, Mahyudin NA et al. (2021) Biocontrol of aflatoxins using non-aflatoxigenic Aspergillus flavus: A literature review. Journal of Fungi (Basel) 7: 381. https://doi.org/10.3390/JOF7050381
- Kjærbølling I, Vesth T, Frisvad JC et al. (2020) A comparative genomics study of 23 Aspergillus species from section Flavi. Nature Communications 11: 1106. https://doi.org/10.1038/S41467-019-14051-Y
- Korschelt O (1878) Über Sake, das alkoholische Getränk der Japaner. Dingler's Polytechnic Journal 230: 76–80.
- Kurtzman CP, Smiley MJ, Robnett CJ et al. (1986) DNA relatedness among wild and domesticated species in the Aspergillus flavus group. Mycologia 78: 955–959. https://doi.org/10.1080/00275514.1986.12025355
- Letunic I, Bork P (2019) Interactive Tree Of Life (iTOL) v4: recent updates and new developments. Nucleic Acids Research: gkz239. https://doi.org/10.1093/nar/gkz239
- Lewis MH, Carbone I, Luis JM et al. (2019) Biocontrol strains differentially shift the genetic structure of indigenous soil populations of Aspergillus flavus. Frontiers in Microbiology 10: 1738. https://doi.org/10.3389/FMICB.2019.01738
- Li H, Durbin R (2010) Fast and accurate long-read alignment with Burrows-Wheeler transform. Bioinformatics 26: 589–595. https://doi.org/10.1093/bioinformatics/btp698
- Link HF (1809) Observationes in ordines plantarum naturales. Dissertatio Ima. Magazin für die Neuesten Entdeckungen in der Gesammten Naturkunde 3: 3–42.
- Loegler V, Thiele P, Teyssonnière E et al. (2025) From genotype to phenotype with 1,086 near telomere-to-telomere yeast genomes. Nature 2025: 1–10. https://doi.org/10.1038/s41586-025-09637-0
- Machida M, Asai K, Sano M et al. (2005) Genome sequencing and analysis of Aspergillus oryzae. Nature 438: 1157–1161. https://doi.org/10.1038/nature04300
- Minh BQ, Schmidt HA, Chernomor O et al. (2020) IQ-TREE 2: New models and efficient methods for phylogenetic inference in the genomic era. Molecular Biology and Evolution 37: 1530–1534. https://doi.org/10.1093/MOLBEV/MSAA015
- Molo MS, White JB, Cornish V et al. (2022) Asymmetrical lineage introgression and recombination in populations of Aspergillus flavus: Implications for biological control. PLoS ONE 17: 1–41. https://doi.org/10.1371/journal.pone.0276556
- Molo MS, Heiniger RW, Boerema L et al. (2019) Trial summary on the comparison of various non-aflatoxigenic strains of aspergillus flavus on mycotoxin levels and yield in maize. Agronomy Journal 111: 942–946. https://doi.org/10.2134/agronj2018.07.0473
- Moore GG, Elliott JL, Singh R et al. (2013) Sexuality generates diversity in the aflatoxin gene cluster: Evidence on a global scale. PLoS Pathogens 9: e1003574. https://doi.org/10.1371/journal.ppat.1003574
- Moore GG, Singh R, Horn BW et al. (2009) Recombination and lineage-specific gene loss in the aflatoxin gene cluster of Aspergillus flavus. Molecular Ecology 18: 4870–4887. https://doi.org/10.1111/j.1365-294X.2009.04414.x
- Moore GG, Olarte RA, Horn BW et al. (2017) Global population structure and adaptive evolution of aflatoxin-producing fungi. Ecology and Evolution 7: 9179–9191. https://doi.org/10.1002/ece3.3464
- Ohkura M, Cotty PJ, Orbach MJ (2018) Comparative genomics of Aspergillus flavus S and L morphotypes yield insights into niche adaptation. G3 (Bethesda) 8: g3.200553.2018. https://doi.org/10.1534/g3.118.200553
- Ortiz EM (2019) vcf2phylip v2.0: convert a VCF matrix into several matrix formats for phylogenetic analysis. https://doi.org/10.5281/ZENODO.2540861
- Peles F, Sipos P, Győri Z et al. (2019) Adverse effects, transformation and channeling of aflatoxins into food raw materials in livestock. Frontiers in Microbiology 10: 2861. https://doi.org/10.3389/fmicb.2019.02861
- Peng Q, Zhou H, Zheng H et al. (2025) Investigating the role of primary fungi in Huangjiu fermentation: Insights from flavor orientation and synthetic microbiomes. Food Microbiology 129: 104765. https://doi.org/10.1016/j.fm.2025.104765
- Peter J, De Chiara M, Friedrich A et al. (2018) Genome evolution across 1,011 Saccharomyces cerevisiae isolates. Nature 556: 339–344. https://doi.org/10.1038/s41586-018-0030-5
- Poplin R, Ruano-Rubio V, DePristo MA et al. (2018) Scaling accurate genetic variant discovery to tens of thousands of samples. bioRxiv 10.1101/201178 https://doi.org/10.1101/201178
- Rácz HV, Imre A, Németh B et al. (2025) Commercial Saccharomyces cerevisiae baker's yeasts: strain redundancy, genome plasticity, and colonization of the sourdough environment and the human body. bioRxiv 10.1101/2025.02.07.637159. https://doi.org/10.1101/2025.02.07.637159
- Raj A, Stephens M, Pritchard JK (2014) FastSTRUCTURE: Variational inference of population structure in large SNP data sets. Genetics 197: 573–589. https://doi.org/10.1534/genetics.114.164350
- Raper KB, Fennell DI (1973) The genus Aspergillus. Robert E. Krieger Co. , Huntington, NY, 686 pp.
- Rasheed U, Cotty PJ, Ain QU et al. (2024) Efficacy of atoxigenic Aspergillus flavus from southern China as biocontrol agents against aflatoxin contamination in corn and peanuts. Pesticide Biochemistry and Physiology 201: 105887. https://doi.org/10.1016/j.pestbp.2024.105887
- Rushing BR, Selim MI (2019) Aflatoxin B1: A review on metabolism, toxicity, occurrence in food, occupational exposure, and detoxification methods. Food and Chemical Toxicology 124: 81–100. https://doi.org/10.1016/j.fct.2018.11.047
- Saito M, Tsuruta O (1993) A new variety of Aspergillus flavus from tropical soil in Thailand and its aflatoxin productivity. JSM Mycotoxins 1993: 31–36. https://doi.org/10.2520/myco1975.1993.31
- Seidler Y, Rimbach G, Lüersen K et al. (2024) The postbiotic potential of Aspergillus oryzae - a narrative review. Frontiers in Microbiology 15: 1452725. https://doi.org/10.3389/FMICB.2024.1452725
- Skerker JM, Pianalto KM, Mondo SJ et al. (2021) Chromosome assembled and annotated genome sequence of Aspergillus flavus NRRL 3357. G3 (Bethesda) 11: jkab213. https://doi.org/10.1093/G3JOURNAL/JKAB213
- StatisticsKingdom (2017) Multiple Linear Regression Calculator. https://www.statskingdom.com/
- Sun Z, Wu Y, Long S et al. (2024) Aspergillus oryzae as a cell factory: Research and applications in industrial production. Journal of Fungi (Basel) 10: 248. https://doi.org/10.3390/JOF10040248
- Uwineza C, Parchami M, Bouzarjomehr M et al. (2024) Recent developments in the application of filamentous fungus Aspergillus oryzae in ruminant feed. Animals 14: 2427. https://doi.org/10.3390/ANI14162427
- Watarai N, Yamamoto N, Sawada K et al. (2019) Evolution of Aspergillus oryzae before and after domestication inferred by large-scale comparative genomic analysis. DNA research : an international journal for rapid publication of reports on genes and genomes 26: 465–472. https://doi.org/10.1093/DNARES/DSZ024
- Xie H, Hu J, Yue X et al. (2023) WITHDRAWN: Global multi-omics profiling reveals evolutionary drivers of phylogeographic diversity of fungal specialized metabolism. ResearchSquare rs-2471999. https://doi.org/10.21203/RS.3.RS-2471999/V1
- Xu J, Zhang Y, Ren J et al. (2025) An atoxigenic Aspergillus flavus PA67 from Shandong province exhibits potential in biocontrol against toxigenic Aspergillus flavus, Sclerotium rolfsii, and Fusarium proliferatum. International Journal of Food Microbiology 426: 110918. https://doi.org/10.1016/J.IJFOODMICRO.2024.110918
- Yan Q, Li S, Yan Q et al. (2024) A genomic compendium of cultivated human gut fungi characterizes the gut mycobiome and its relevance to common diseases. Cell 187: 2969-2989.e24. https://doi.org/10.1016/J.CELL.2024.04.043
- You Y, Xu X, Liu H et al. (2024) Locust pathogen Aspergillus oryzae XJ1 is different from Aspergillus oryzae and Aspergillus flavus based on genomics comparisons. Microorganisms 12: 2501. https://doi.org/10.3390/MICROORGANISMS12122501
- Yun J, Kim JH, Lee JE (2020) Influence of aflatoxin in Nuruk on the safety of starch-based alcoholic beverage. Journal of Food Science 85: 762–770. https://doi.org/10.1111/1750-3841.15050
- Zheng X, Levine D, Shen J et al. (2012) A high-performance computing toolset for relatedness and principal component analysis of SNP data. Bioinformatics 28: 3326–3328. https://doi.org/10.1093/BIOINFORMATICS/BTS606