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      Comparing genotype and chemotype of Fusarium graminearum from cereals in Ontario, Canada

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          Abstract

          Fusarium graminearum is responsible for production of the mycotoxin deoxynivalenol (DON) on maize and wheat in Ontario, Canada. It has been understood since the early 1980s that in most parts of Canada, the predominant chemotype of F. graminearum is 15ADON, and not the 3ADON chemotype mainly found in Europe and Asia. The discovery of F. graminearum strains that did not produce DON but the structurally related 7-α hydroxy, 15-deacetylcalonectrin (3ANX) and its hydrolysis product 7-α hydroxy, 3,15-dideacetylcalonectrin to (NX) demonstrated that we still have a lot to learn about this well studied but complicated fungus. We conducted a survey of maize and wheat samples from Ontario farms. In the 2015 crop year, we isolated 86 strains and tested a representative subset of 20 using the published genetic probes for assessing genotype. We also developed a targeted LC-MS/MS method for the identification and quantitation of known toxins from this species to determine chemotype. The results showed that 80% of our strains produced some 3ANX in addition to 15ADON and one strain produced 3ANX and no 15ADON. Comparison of chemical data with genotyping revealed that in more than 50% of the cases there was no clear agreement. These data demonstrate the importance of chemical analysis for understanding the toxigenic potential of strains, especially using a LC-MS method that is capable of differentiating 3ADON and 15ADON. For this collection, genotyping of isolates did not produce reliable information on the chemotype. This is the first report of 3ANX toxin production concurrently with 15ADON and suggests that the 3ANX producers in North America likely originated from the 15ADON background.

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          Gene genealogies reveal global phylogeographic structure and reproductive isolation among lineages of Fusarium graminearum, the fungus causing wheat scab.

          During the past decade, the plant disease called scab or Fusarium head blight of wheat and barley has reached epidemic proportions in North America and elsewhere in the world. Scab is an economically devastating plant disease, not only because it causes significant reduction in seed yields and quality, but also because infested seeds are often contaminated with trichothecene and estrogenic mycotoxins that pose a serious threat to animal health and food safety. To test whether the primary etiological agent of scab, the fungus Fusarium graminearum, is panmictic throughout its range, allelic genealogies were constructed from six single-copy nuclear genes from strains selected to represent the global genetic diversity of this pathogen. Excluding one hybrid strain, all six genealogies recovered the same seven biogeographically structured lineages, suggesting that they represent phylogenetically distinct species among which gene flow has been very limited during their evolutionary history. Parsimony analysis of the combined data set comprising 7,120 aligned nucleotide characters resolved most relationships among the seven lineages of the F. graminearum clade and related fusaria included in the study. Phylogenetic evidence is also presented for introgressive hybridization and intragenic recombination among lineages of the F. graminearum clade in nature.
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            An adaptive evolutionary shift in Fusarium head blight pathogen populations is driving the rapid spread of more toxigenic Fusarium graminearum in North America.

            Analysis of Fusarium head blight (FHB) pathogen diversity revealed that 3ADON producing Fusarium graminearum are prevalent in North America and identified significant population structure associated with trichothecene chemotype differences (F(ST)>0.285; P<0.001). In addition, we identified a trichothecene chemotype cline in Canada and documented a recent and significant shift in FHB pathogen composition by demonstrating that the 3ADON chemotype frequency in western Canada increased more than 14-fold between 1998 and 2004. On average, isolates from 3ADON populations produced significantly (P<0.05) more trichothecene and had significantly (P<0.005) higher fecundity and growth rates than isolates from the 15ADON population. These results indicate that selection is driving the rapid spread of an introduced pathogen population that is more toxigenic and potentially more vigorous. The discovery of this previously unrecognized pathogen diversity has significant implications for food safety and cereal production in North America.
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              Global molecular surveillance reveals novel Fusarium head blight species and trichothecene toxin diversity.

              To expand our knowledge of Fusarium head blight (FHB) pathogen and trichothecene toxin diversity, a global collection of 2100 isolates was screened for novel genetic variation, resulting in the identification of 16 phylogenetically divergent FHB isolates. The affinities and taxonomic status of these novel isolates were evaluated via phylogenetic analyses of multilocus DNA sequence data (13 genes; 16.3 kb/strain) together with analyses of their morphology, pathogenicity to wheat, and trichothecene toxin potential. Based on the results of these analyses, we formally describe two novel species (Fusarium vorosii and Fusarium gerlachii) within the Fusarium graminearum species complex (Fg complex), and provide the first published report of Fg complex isolates with either a nivalenol or 3-acetyldeoxynivalenol chemotype within the U.S. In addition, we describe a highly divergent population of F. graminearum from the Gulf Coast of the U.S., and divergent isolates of F. acaciae-mearnsii from Australia and South Africa.
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                Author and article information

                Contributors
                Role: InvestigationRole: Writing – review & editing
                Role: InvestigationRole: MethodologyRole: Writing – original draft
                Role: InvestigationRole: Writing – review & editing
                Role: ConceptualizationRole: Funding acquisitionRole: InvestigationRole: MethodologyRole: Writing – original draft
                Role: Editor
                Journal
                PLoS One
                PLoS ONE
                plos
                plosone
                PLoS ONE
                Public Library of Science (San Francisco, CA USA )
                1932-6203
                9 May 2019
                2019
                : 14
                : 5
                : e0216735
                Affiliations
                [1 ] Department of Chemistry, Carleton University, Ottawa, Ontario, Canada
                [2 ] London Research and Development Center, Agriculture and Agri-Food Canada, London, Canada
                Universita degli Studi di Pisa, ITALY
                Author notes

                Competing Interests: The authors have declared that no competing interests exist.

                Author information
                http://orcid.org/0000-0002-6680-6563
                Article
                PONE-D-18-36518
                10.1371/journal.pone.0216735
                6508712
                31071188
                29054ef0-6f9f-42f9-9bf8-529e40602714
                © 2019 Crippin et al

                This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

                History
                : 21 December 2018
                : 26 April 2019
                Page count
                Figures: 5, Tables: 2, Pages: 18
                Funding
                Funded by: funder-id http://dx.doi.org/10.13039/501100000094, Ontario Ministry of Agriculture, Food and Rural Affairs;
                Award ID: FS2015-2617 / SF6115
                Award Recipient :
                J David Miller funded by the Ontario Ministry of Food and Agriculture (FS2015-2617 / SF6115). This is the Department of Agriculture for the Province of Ontario, Canada. The funders played no role in the design or execution of the project.
                Categories
                Research Article
                Research and Analysis Methods
                Animal Studies
                Experimental Organism Systems
                Model Organisms
                Maize
                Research and Analysis Methods
                Model Organisms
                Maize
                Biology and Life Sciences
                Organisms
                Eukaryota
                Plants
                Grasses
                Maize
                Research and Analysis Methods
                Animal Studies
                Experimental Organism Systems
                Plant and Algal Models
                Maize
                Biology and Life Sciences
                Organisms
                Eukaryota
                Plants
                Grasses
                Wheat
                Biology and Life Sciences
                Molecular Biology
                Molecular Biology Techniques
                Artificial Gene Amplification and Extension
                Polymerase Chain Reaction
                Research and Analysis Methods
                Molecular Biology Techniques
                Artificial Gene Amplification and Extension
                Polymerase Chain Reaction
                Biology and Life Sciences
                Toxicology
                Toxic Agents
                Toxins
                Medicine and Health Sciences
                Pathology and Laboratory Medicine
                Toxicology
                Toxic Agents
                Toxins
                Physical Sciences
                Chemistry
                Analytical Chemistry
                Chemical Analysis
                Biology and Life Sciences
                Genetics
                Fungal Genetics
                Biology and Life Sciences
                Mycology
                Fungal Genetics
                Biology and Life Sciences
                Agriculture
                Farms
                Biology and Life Sciences
                Microbiology
                Medical Microbiology
                Microbial Pathogens
                Fungal Pathogens
                Fusarium
                Fusarium Graminearum
                Medicine and Health Sciences
                Pathology and Laboratory Medicine
                Pathogens
                Microbial Pathogens
                Fungal Pathogens
                Fusarium
                Fusarium Graminearum
                Biology and Life Sciences
                Mycology
                Fungal Pathogens
                Fusarium
                Fusarium Graminearum
                Custom metadata
                Relevant data underlying the study is available within the Supporting Information file, including GenBank accession numbers for all strains.

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                Uncategorized

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