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

ABSTRACT: Eucalyptus 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 × 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.

SUBMITTER: Elorriaga E

PROVIDER: S-EPMC8428835 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:Many clonal plants have two reproductive patterns, seed propagation and vegetative propagation. By vegetative propagation, plants reproduce the genetically identical offspring with a low mortality, because resources are supplied from the other individuals through interconnected ramets at vegetative-propagated offspring. However, the ramets transport not only resources but also systemic pathogen. Pathogens evolve to establish and spread widely within the plant population. The superinfection, which is defined as the ability that an established pathogen spreads widely by infecting to already-infected individuals with other strains of a pathogen, is important to the evolution of pathogens. We examine the dynamics of plant reproduction and pathogen propagation considering spatial structure and the effect of superinfection on genetic diversity of pathogen by analysis of several models, 1-strain and multiple-strain models, on two-dimensional square lattice. In the analysis of 1-strain model, we derive equilibrium value by mean-field approximation and pair approximation, and its local stability by Routh-Hurwitz stability criterion. In the multiple-strain models, we analyze the dynamics by numerical simulation of mean-field approximation, pair approximation and Monte Carlo simulation. Through the analyses, we show the effect of parameter values to dynamics of models, such as transition of dominant strain of pathogen, competition between plants and pathogens and density of individuals. As a result, (i) The strain with intermediate cost becomes dominant when both superinfection rate and growth rate are low. (ii) The competition between plants and pathogens occurs in the phase of coexistence of various strains by pair approximation and Monte Carlo simulation. (iii) Too high growth rate leads to the decrease of plant population in all models. (iv) Pathogens are easy to maintain their genetic diversity with low superinfection rate. However, if they do not superinfect, the maintenance becomes difficult. (v) When growth rate of plant is low, individuals are very influenced by distant individuals.

Dataset Information

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

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