Genome Editing to Produce Knockout Mutations of Seed Dormancy Genes in Wheat

Fumitaka Abe; Yoko Kamiya; Yuji Ishida; Hiroshi Hisano; Kanako Kawaura; Toshihiko Komari; Kazuhiro Sato

doi:10.1007/978-1-0716-3965-8_13

Genome Editing to Produce Knockout Mutations of Seed Dormancy Genes in Wheat

Methods Mol Biol. 2024:2830:137-148. doi: 10.1007/978-1-0716-3965-8_13.

Authors

Fumitaka Abe¹, Yoko Kamiya², Yuji Ishida³, Hiroshi Hisano⁴, Kanako Kawaura², Toshihiko Komari³, Kazuhiro Sato^{4

5

6}

Affiliations

¹ Institute of Crop Science, NARO, Tsukuba, Japan. [email protected].
² Kihara Institute for Biological Research, Yokohama City University, Yokohama, Japan.
³ Plant Innovation Center, Japan Tobacco Inc. (currently Agri-Bio Research Center, KANEKA CORPORATION), Iwata, Japan.
⁴ Institute of Plant Science & Resources, Okayama University, Kurashiki, Japan.
⁵ Faculty of Agriculture, Setsunan University, Hirakata, Japan.
⁶ Kazusa DNA research Institute, Kisarazu, Japan.

PMID: 38977575
DOI: 10.1007/978-1-0716-3965-8_13

Abstract

Knockout mutants provide definitive information about the functions of genes related to agronomic traits, including seed dormancy. However, it takes many years to produce knockout mutants using conventional techniques in polyploid plants such as hexaploid wheat. Genome editing with sequence-specific nucleases is a promising approach for obtaining knockout mutations in all targeted homoeologs of wheat simultaneously. Here, we describe a procedure to produce a triple recessive mutant in wheat via genome editing. This protocol covers the evaluation of gRNA and Agrobacterium-mediated transformation to obtain edited wheat seedlings.

Keywords: Agrobacterium-mediated transformation; CRISPR/Cas9; Genome editing; Immature embryo; Wheat.

MeSH terms

Agrobacterium / genetics
CRISPR-Cas Systems*
Gene Editing* / methods
Gene Knockout Techniques* / methods
Genes, Plant
Genome, Plant
Mutation
Plant Dormancy* / genetics
Plants, Genetically Modified / genetics
RNA, Guide, CRISPR-Cas Systems / genetics
Seedlings / genetics
Seeds / genetics
Triticum* / genetics

Substances

RNA, Guide, CRISPR-Cas Systems