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Plant Biotechnology Journal - 2022 - Hyde - Identification characterization and rescue of CRISPR Cas9 generated wheat.pdf (3.42 MB)

Identification, characterization, and rescue of CRISPR/Cas9 generated wheat SPO11-1 mutants

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posted on 2023-05-16, 11:20 authored by Lucy Hyde, Kim Osman, Mark Winfield, Eugenio Sanchez‐Moran, James D Higgins, Ian R Henderson, Caroline Sparks, F Chris H Franklin, Keith J Edwards

Increasing crop yields through plant breeding is time consuming and laborious, with the generation of novel combinations of alleles being limited by chromosomal linkage blocks and linkage-drag. Meiotic recombination is essential to create novel genetic variation via the reshuffling of parental alleles. The exchange of genetic information between homologous chromosomes occurs at crossover (CO) sites but CO frequency is often low and unevenly distributed. This bias creates the problem of linkage-drag in recombination ‘cold’ regions, where undesirable variation remains linked to useful traits. In plants, programmed meiosis-specific DNA double-strand breaks, catalysed by the SPO11 complex, initiate the recombination pathway, although only ~5% result in the formation of COs. To study the role of SPO11-1 in wheat meiosis, and as a prelude to manipulation, we used CRISPR/Cas9 to generate edits in all three SPO11-1 homoeologues of hexaploid wheat. Characterization of progeny lines shows plants deficient in all six SPO11-1 copies fail to undergo chromosome synapsis, lack COs and are sterile. In contrast, lines carrying a single copy of any one of the three wild-type homoeologues are phenotypically indistinguishable from unedited plants both in terms of vegetative growth and fertility. However, cytogenetic analysis of the edited plants suggests that homoeologues differ in their ability to generate COs and in the dynamics of synapsis. In addition, we show that the transformation of wheat mutants carrying six edited copies of SPO11-1 with the TaSPO11-1B gene, restores synapsis, CO formation, and fertility and hence opens a route to modifying recombination in this agronomically important crop.

Funding

Investigating the role of the meiotic chromosome axes in mediating crossover designation in bread wheat

Biotechnology and Biological Sciences Research Council

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A pipeline for efficient recombination in wheat

Biotechnology and Biological Sciences Research Council

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History

Author affiliation

Department of Genetics and Genome Biology, University of Leicester

Version

  • VoR (Version of Record)

Published in

Plant Biotechnology Journal

Volume

21

Issue

2

Pagination

405 - 418

Publisher

Wiley

issn

1467-7644

eissn

1467-7652

Copyright date

2023

Available date

2023-05-16

Language

en

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