Application of a GRF-GIF chimera enhances plant regeneration for genome editing in tomato

2024 · doi:10.1101/2024.12.20.628945
preprint OA: gold CC-BY-NC-ND-4.0
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Expressing a tomato GRF-GIF chimera accelerates shoot regeneration, increases transgenic plant recovery, and simplifies mutant isolation for genome editing in tomato.

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This paper studied whether expressing a morphogenic regulator chimera, GRF-GIF, could improve plant regeneration and genome editing efficiency in tomato, using tomato tissue culture and genome editing workflows to measure shoot regeneration timing and recovery of heritable edited transgenic plants. The authors found that the tomato GRF-GIF chimera accelerated shoot regeneration from callus tissue culture by about one month and nearly doubled the number of recovered transgenic plants, leading to recovery of a broader range of edited haplotypes and facilitating isolation of mutants with heritable edits, while not markedly disrupting plant growth and development. The main limitation is that the work is framed as a technical improvement in tomato transformation rather than as a biological mechanism study of regeneration itself. The paper does not explicitly discuss endometriosis or adenomyosis; it was included in the corpus via a keyword match in the upstream search index.

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Abstract

Genome editing has become a routine tool for functionally characterizing plant and animal genomes. However, stable genome editing in plants remains limited by the time- and labor- intensive process of generating transgenic plants, as well as by the efficient isolation of desired heritable edits. In this study, we evaluated the impact of the morphogenic regulator GRF-GIF on plant regeneration and genome editing outcomes in tomato. We demonstrate that expressing a tomato GRF-GIF chimera reliably accelerates the onset of shoot regeneration from callus tissue culture by approximately one month and nearly doubles the number of recovered transgenic plants. Consequently, the GRF-GIF chimera enables the recovery of a broader range of edited haplotypes and simplifies the isolation of mutants harboring heritable edits, but without markedly interfering with plant growth and development. Based on these findings, we outline strategies that employ basic or advanced diagnostic pipelines for efficient isolation of single and higher-order mutants in tomato. Our work represents a technical advantage for tomato transformation and genome editing, with potential applications across other Solanaceae species.
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Abstract Genome editing has become a routine tool for functionally characterizing plant and animal genomes. However, stable genome editing in plants remains limited by the time- and labor- intensive process of generating transgenic plants, as well as by the efficient isolation of desired heritable edits. In this study, we evaluated the impact of the morphogenic regulator GRF-GIF on plant regeneration and genome editing outcomes in tomato. We demonstrate that expressing a tomato GRF-GIF chimera reliably accelerates the onset of shoot regeneration from callus tissue culture by approximately one month and nearly doubles the number of recovered transgenic plants. Consequently, the GRF-GIF chimera enables the recovery of a broader range of edited haplotypes and simplifies the isolation of mutants harboring heritable edits, but without markedly interfering with plant growth and development. Based on these findings, we outline strategies that employ basic or advanced diagnostic pipelines for efficient isolation of single and higher-order mutants in tomato. Our work represents a technical advantage for tomato transformation and genome editing, with potential applications across other Solanaceae species. Competing Interest Statement The authors have declared no competing interest.

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License: CC-BY-NC-ND-4.0