CRISPR-Cas9 Targeted Mutagenesis Leads to Simultaneous Modification of Different Homoeologous Gene Copies in Polyploid Oilseed Rape ( Brassica napus )

In polyploid species, altering a trait by random mutagenesis is highly inefficient due to gene redundancy. We have stably transformed tetraploid oilseed rape ( ) with a CRISPR-Cas9 construct targeting two ( ) homoeologs. is involved in valve margin development and, thus, contributes to seed shatteri...

Celý popis

Uložené v:
Podrobná bibliografia
Vydané v:Plant physiology (Bethesda) Ročník 174; číslo 2; s. 935
Hlavní autori: Braatz, Janina, Harloff, Hans-Joachim, Mascher, Martin, Stein, Nils, Himmelbach, Axel, Jung, Christian
Médium: Journal Article
Jazyk:English
Vydavateľské údaje: United States 01.06.2017
Predmet:
Base Sequence
Brassica napus - genetics
Brassica napus - growth & development
CRISPR-Cas Systems - genetics
DNA, Bacterial - genetics
Gene Dosage
Gene Targeting
Genome, Plant
Inheritance Patterns - genetics
Mutagenesis, Insertional - genetics
Mutation - genetics
Polyploidy
Seeds - genetics
Sequence Homology, Nucleic Acid
Transformation, Genetic
ISSN:1532-2548, 1532-2548
On-line prístup:Zistit podrobnosti o prístupe
Tagy: Pridať tag
Žiadne tagy, Buďte prvý, kto otaguje tento záznam!
Popis
Shrnutí:In polyploid species, altering a trait by random mutagenesis is highly inefficient due to gene redundancy. We have stably transformed tetraploid oilseed rape ( ) with a CRISPR-Cas9 construct targeting two ( ) homoeologs. is involved in valve margin development and, thus, contributes to seed shattering from mature fruits. Knocking out would increase shatter resistance to avoid seed loss during mechanical harvest. We obtained a transgenic T1 plant with four mutant alleles by the use of a single target sequence. All mutations were stably inherited to the T2 progeny. The T2 generation was devoid of any wild-type alleles, proving that the underlying T1 was a nonchimeric double heterozygote. T-DNA and loci were not linked, as indicated by random segregation in the T2 generation. Hence, we could select double mutants lacking the T-DNA already in the first offspring generation. However, whole-genome sequencing data revealed at least five independent insertions of vector backbone sequences. We did not detect any off-target effects in two genome regions homologous to the target sequence. The simultaneous alteration of multiple homoeologs by CRISPR-Cas9 mutagenesis without any background mutations will offer new opportunities for using mutant genotypes in rapeseed breeding.
Bibliografia:ObjectType-Article-1
SourceType-Scholarly Journals-1
ObjectType-Feature-2
content type line 23
ISSN:1532-2548
1532-2548
DOI:10.1104/pp.17.00426