Targeted Gene Mutations in the Forest Pathogen Dothistroma septosporum Using CRISPR/Cas9.

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dc.citation.issue8
dc.citation.volume11
dc.contributor.authorMcCarthy HM
dc.contributor.authorTarallo M
dc.contributor.authorMesarich CH
dc.contributor.authorMcDougal RL
dc.contributor.authorBradshaw RE
dc.coverage.spatialSwitzerland
dc.date.available2022-04-08
dc.date.available2022-04-04
dc.date.issued2022-04-08
dc.description.abstractDothistroma needle blight, caused by Dothistroma septosporum, has increased in incidence and severity over the last few decades and is now one of the most important global diseases of pines. Disease resistance breeding could be accelerated by knowledge of pathogen virulence factors and their host targets. However, this is hindered due to inefficient targeted gene disruption in D. septosporum, which is required for virulence gene characterisation. Here we report the first successful application of CRISPR/Cas9 gene editing to a Dothideomycete forest pathogen, D. septosporum. Disruption of the dothistromin pathway regulator gene AflR, with a known phenotype, was performed using nonhomologous end-joining repair with an efficiency of > 90%. Transformants with a range of disruption mutations in AflR were produced. Disruption of Ds74283, a D. septosporum gene encoding a secreted cell death elicitor, was also achieved using CRISPR/Cas9, by using a specific donor DNA repair template to aid selection where the phenotype was unknown. In this case, 100% of screened transformants were identified as disruptants. In establishing CRISPR/Cas9 as a tool for gene editing in D. septosporum, our research could fast track the functional characterisation of candidate virulence factors in D. septosporum and helps set the foundation for development of this technology in other forest pathogens.
dc.description.publication-statusPublished online
dc.identifierhttps://www.ncbi.nlm.nih.gov/pubmed/35448744
dc.identifierplants11081016
dc.identifier.citationPlants (Basel), 2022, 11 (8)
dc.identifier.doi10.3390/plants11081016
dc.identifier.elements-id452893
dc.identifier.harvestedMassey_Dark
dc.identifier.issn2223-7747
dc.languageeng
dc.relation.isPartOfPlants (Basel)
dc.rights(c) The author/s (CC BY)
dc.subjectCRISPR/Cas9
dc.subjectDSB repair
dc.subjectDothideomycete
dc.subjectDothistroma
dc.subjectDothistroma needle blight
dc.subjectfungal transformation
dc.subjectgene disruption
dc.subjecthomologous recombination
dc.subjectpine pathogen
dc.titleTargeted Gene Mutations in the Forest Pathogen Dothistroma septosporum Using CRISPR/Cas9.
dc.typeJournal article
pubs.notesNot known
pubs.organisational-group/Massey University
pubs.organisational-group/Massey University/College of Sciences
pubs.organisational-group/Massey University/College of Sciences/School of Agriculture & Environment
pubs.organisational-group/Massey University/College of Sciences/School of Natural Sciences
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