Efficient targeted mutagenesis in allotetraploid sweet basil by CRISPR/Cas9

Plant Direct
Natasha Navet, Miaoying Tian

Abstract

Sweet basil (Ocimum basilicum) is an economically important herb and its global production is threatened by basil downy mildew caused by the obligate biotrophic oomycete Peronospora belbahrii. Effective tools are required for functional understanding of its genes involved in synthesis of valuable secondary metabolites in essential oil and disease resistance, and breeding for varieties with improved traits. Clustered regularly interspaced short palindromic repeat (CRISPR)/Cas9 gene editing technology has revolutionized crop breeding and functional genomics. The applicability and efficacy of this genomic tool in the allotetraploid sweet basil were tested by editing a potential susceptibility (S) gene ObDMR1, the basil homolog of Arabidopsis DMR1 (Downy Mildew Resistant 1) whose mutations conferred nearly complete resistance against Arabidopsis downy mildew pathogen, Hyaloperonospora arabidopsidis. Two single guide RNAs targeting two different sites of the ObDMR1 coding sequence were designed. A total of 56 transgenic lines were obtained via Agrobacterium-mediated stable transformation. Mutational analysis of 54 T0 transgenic lines identified 92.6% lines carrying mutations at target 1 site, while a very low mutation frequency was ...Continue Reading

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Citations

Dec 20, 2020·DNA Research : an International Journal for Rapid Publication of Reports on Genes and Genomes·Itay GondaNativ Dudai
Apr 10, 2021·Journal of Zhejiang University. Science. B·Chao LiBaohong Zhang
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Aug 28, 2021·Frontiers in Plant Science·Narayan Chandra PaulHyunkyu Sang
Nov 17, 2021·The Plant Journal : for Cell and Molecular Biology·Juvenal AssouJens Boch

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Datasets Mentioned

BETA
MT000722

Methods Mentioned

BETA
transgenic
PCR
ICE
reverse

Software Mentioned

RNAstructure
Illumina Control
Synthego ICE
Conserved Domain Search
EuPaGDT
Genoverser
CLUSTALX
BOXSHADE
SAS
TBLASTX

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