Research achievements (PUBLICATIONS)

Basic information

Name KOIZUKA Nobuya

Name of dissertation

CRISPR/Cas9-mediated genome editing of the fatty acid desaturase 2 gene in Brassica napus

Name of author

Ayako Okuzakia, TakumiOgawa, Chie Koizuka 0Kanako Kanekoa, Mizue Inaba, Jun Imamura and Nobuya Koizuka*

Name of publication

Plant Physiology and Biochemistry

Publisher

Elsevier

Volume (No.)

 

Issue (No.)

 

Start page

 

 

 

End page

 

Publication date

2018

Presence/absence of peer review

maru

Presence/absence of invitation

Batu

Language of publication used

 

Posting category

 

Type of publication

 

Type of authorship

Joint Work(other)

ISSN

 

DOI

10.1016/j.plaphy.2018.04.025

NAID (CiNii ID)

 

PMID

 

Permalink

URL

Summary

We report the modification of a fatty acid desaturase 2 gene (FAD2), which encodes an enzyme that catalyzes the desaturation of oleic acid, in Brassica napus cv. Westar using the CRISPR/Cas9 system. Two guide RNAs were designed for BnaA.FAD2.a (FAD2_Aa). Of 22 regenerated shoots with FAD2_Aa editing vectors, three contained mutant alleles. Further analysis revealed that two of three mature plants (Aa1#13 and Aa2#2) contained the mutant alleles. The mutant fad2_Aa allele had a 4-bp deletion, which was inherited by backcross progenies (BC1) in the Aa1#13 line. Furthermore, plants with the fad2_Aa allele without transgenes were selected from the BC1 progenies and plants homozygous for fad2_Aa were then produced by self-crossing these BC1 progenies (BC1S1). Fatty acid composition analysis of their seeds revealed a statistically significant increase in the content of oleic acid compared with that in wild-type seeds. These results showed that the application of the CRISPR/Cas9 system is useful to produce desirable mutant plants with an agronomically suitable phenotype by modifying the metabolic pathway in B. napus.

Remarks