Genetic containment in vegetatively propagated forest trees : CRISPR disruption of LEAFY function in Eucalyptus gives sterile indeterminate inflorescences and normal juvenile development

dc.contributor.authorElorriaga, Estefania
dc.contributor.authorKlocko, Amy L.
dc.contributor.authorMa, Cathleen
dc.contributor.authorDu Plessis, Marc
dc.contributor.authorAn, Xinmin
dc.contributor.authorMyburg, Alexander Andrew
dc.contributor.authorStrauss, Steven H.
dc.date.accessioned2022-03-10T10:48:08Z
dc.date.available2022-03-10T10:48:08Z
dc.date.issued2021-09
dc.description.abstractEucalyptus is among the most widely planted taxa of forest trees worldwide. However, its spread as an exotic or genetically engineered form can create ecological and social problems. To mitigate gene flow via pollen and seeds, we mutated the Eucalyptus orthologue of LEAFY (LFY) by transforming a Eucalyptus grandis 9 urophylla wild-type hybrid and two Flowering Locus T (FT) overexpressing (and flowering) lines with CRISPR Cas9 targeting its LFY orthologue, ELFY. We achieved high rates of elfy biallelic knockouts, often approaching 100% of transgene insertion events. Frameshift mutations and deletions removing conserved amino acids caused strong floral alterations, including indeterminacy in floral development and an absence of male and female gametes. These mutants were otherwise visibly normal and did not differ statistically from transgenic controls in juvenile vegetative growth rate or leaf morphology in greenhouse trials. Genes upstream or near to ELFY in the floral development pathway were overexpressed, whereas floral organ identity genes downstream of ELFY were severely depressed. We conclude that disruption of ELFY function appears to be a useful tool for sexual containment, without causing statistically significant or large adverse effects on juvenile vegetative growth or leaf morphology.en_ZA
dc.description.departmentBiochemistryen_ZA
dc.description.departmentForestry and Agricultural Biotechnology Institute (FABI)en_ZA
dc.description.departmentGeneticsen_ZA
dc.description.departmentMicrobiology and Plant Pathologyen_ZA
dc.description.departmentZoology and Entomologyen_ZA
dc.description.librarianpm2022en_ZA
dc.description.urihttp://onlinelibrary.wiley.com/journal/10.1111/(ISSN)1467-7652en_ZA
dc.identifier.citationElorriaga E., Klocko A.L., Ma C., du Plessis M., An X., Myburg A.A., Strauss S.H. Genetic containment in vegetatively propagated forest trees: CRISPR disruption of LEAFY function in Eucalyptus gives sterile indeterminate inflorescences and normal juvenile development. Plant Biotechnology Journal 2021 Sep;19(9):1743-1755. doi: 10.1111/pbi.13588.en_ZA
dc.identifier.issn1467-7644 (print)
dc.identifier.issn1467-7652(online)
dc.identifier.other10.1111/pbi.13588
dc.identifier.urihttp://hdl.handle.net/2263/84423
dc.language.isoenen_ZA
dc.publisherWiley Open Accessen_ZA
dc.rights© 2021 The Authors. Plant Biotechnology Journal published by Society for Experimental Biology and The Association of Applied Biologists and John Wiley & Sons Ltd. This is an open access article under the terms of the Creative Commons Attribution License.en_ZA
dc.subjectLEAFYen_ZA
dc.subjectEucalyptusen_ZA
dc.subjectFloweringen_ZA
dc.subjectCRISPRen_ZA
dc.subjectContainmenten_ZA
dc.titleGenetic containment in vegetatively propagated forest trees : CRISPR disruption of LEAFY function in Eucalyptus gives sterile indeterminate inflorescences and normal juvenile developmenten_ZA
dc.typeArticleen_ZA

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