Protocol for Genome Editing to Produce Multiple Mutants in Wheat

Protocol for Genome Editing to Produce Multiple Mutants in Wheat

Summary: 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...

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Main Authors: Fumitaka Abe, Yuji Ishida, Hiroshi Hisano, Masaki Endo, Toshihiko Komari, Seiichi Toki, Kazuhiro Sato
Format: Article
Language:English
Published: Elsevier 2020-09-01
Series:STAR Protocols
Online Access:http://www.sciencedirect.com/science/article/pii/S266616672030040X
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spelling doaj-e974e932a71e44c990446711fc83cb762020-11-25T04:00:53ZengElsevierSTAR Protocols2666-16672020-09-0112100053Protocol for Genome Editing to Produce Multiple Mutants in WheatFumitaka Abe0Yuji Ishida1Hiroshi Hisano2Masaki Endo3Toshihiko Komari4Seiichi Toki5Kazuhiro Sato6Division of Basic Research, Institute of Crop Science, NARO, Tsukuba 305-8518, Japan; Corresponding authorPlant Innovation Center, Japan Tobacco Inc., Iwata 438-0802, JapanInstitute of Plant Science and Resources, Okayama University, Kurashiki 710-0046, JapanDivision of Applied Genetics, Institute of Agrobiological Sciences, NARO, Tsukuba 305-8634, JapanPlant Innovation Center, Japan Tobacco Inc., Iwata 438-0802, JapanDivision of Applied Genetics, Institute of Agrobiological Sciences, NARO, Tsukuba 305-8634, JapanInstitute of Plant Science and Resources, Okayama University, Kurashiki 710-0046, JapanSummary: 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).http://www.sciencedirect.com/science/article/pii/S266616672030040X
collection DOAJ
language English
format Article
sources DOAJ
author Fumitaka Abe
Yuji Ishida
Hiroshi Hisano
Masaki Endo
Toshihiko Komari
Seiichi Toki
Kazuhiro Sato
spellingShingle Fumitaka Abe
Yuji Ishida
Hiroshi Hisano
Masaki Endo
Toshihiko Komari
Seiichi Toki
Kazuhiro Sato
Protocol for Genome Editing to Produce Multiple Mutants in Wheat
STAR Protocols
author_facet Fumitaka Abe
Yuji Ishida
Hiroshi Hisano
Masaki Endo
Toshihiko Komari
Seiichi Toki
Kazuhiro Sato
author_sort Fumitaka Abe
title Protocol for Genome Editing to Produce Multiple Mutants in Wheat
title_short Protocol for Genome Editing to Produce Multiple Mutants in Wheat
title_full Protocol for Genome Editing to Produce Multiple Mutants in Wheat
title_fullStr Protocol for Genome Editing to Produce Multiple Mutants in Wheat
title_full_unstemmed Protocol for Genome Editing to Produce Multiple Mutants in Wheat
title_sort protocol for genome editing to produce multiple mutants in wheat
publisher Elsevier
series STAR Protocols
issn 2666-1667
publishDate 2020-09-01
description Summary: 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).
url http://www.sciencedirect.com/science/article/pii/S266616672030040X
work_keys_str_mv AT fumitakaabe protocolforgenomeeditingtoproducemultiplemutantsinwheat
AT yujiishida protocolforgenomeeditingtoproducemultiplemutantsinwheat
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