Protocol for Genome Editing to Produce Multiple Mutants in Wheat

Fumitaka Abe; Yuji Ishida; Hiroshi Hisano; Masaki Endo; Toshihiko Komari; Seiichi Toki; Kazuhiro Sato

doi:10.1016/j.xpro.2020.100053

Protocol for Genome Editing to Produce Multiple Mutants in Wheat

STAR Protoc. 2020 Jun 13;1(2):100053. doi: 10.1016/j.xpro.2020.100053. eCollection 2020 Sep 18.

Authors

Fumitaka Abe¹, Yuji Ishida², Hiroshi Hisano³, Masaki Endo⁴, Toshihiko Komari², Seiichi Toki⁴, Kazuhiro Sato³

Affiliations

¹ Division of Basic Research, Institute of Crop Science, NARO, Tsukuba 305-8518, Japan.
² Plant Innovation Center, Japan Tobacco Inc., Iwata 438-0802, Japan.
³ Institute of Plant Science and Resources, Okayama University, Kurashiki 710-0046, Japan.
⁴ Division of Applied Genetics, Institute of Agrobiological Sciences, NARO, Tsukuba 305-8634, Japan.

Abstract

Here, we describe a protocol for producing multiple recessive mutants via genome editing in hexaploid wheat (Triticum aestivum) cv. Fielder. Using Agrobacterium-delivered CRISPR/Cas9 and three sub-genome-specific primer sets, all possible combinations of single, double, and triple transgene-free mutants can be generated. The technique for acceleration of generation advancement with embryo culture reduces time for mutant production. The mutants produced by this protocol can be used for the analysis of gene function and crop improvement. For complete details on the use and execution of this protocol, please refer to Abe et al. (2019).

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

CRISPR-Cas Systems / genetics
Gene Editing / methods*
Genome, Plant / genetics*
Mutation / genetics*
Plant Breeding / methods*
Triticum / genetics*