CRISPR/Cas9‐mediated efficient targeted mutagenesis in grape in the first generation

• DOI: 10.1111/pbi.12832
• Publication Date: 2017-09-14T08:01:41Z
• Authors: Xianhang Wang; Mingxing Tu; Dejun Wang; Jianwei Liu; Yajuan Li; Zhi Li; Yuejin Wang; Xiping Wang
• Keywords: Gene Editing; 0301 basic medicine; 2. Zero hunger; Genetic Vectors; Homozygote; Plants, Genetically Modified; Gene Knockout Techniques; Plant Breeding; 03 medical and health sciences; Mutagenesis; Mutation; Mutagenesis, Site-Directed; Vitis; Botrytis; CRISPR-Cas Systems; Research Articles; Disease Resistance; Plant Diseases; Plant Proteins; Transcription Factors

The clustered regularly interspaced short palindromic repeats-associated protein 9 (CRISPR/Cas9) system is a powerful tool for editing plant genomes. Efficient genome of grape (Vitis vinifera) suspension cells using the type II CRISPR/Cas9 has been demonstrated; however, it not established whether this can be applied to get biallelic mutations in first generation grape. In current study, we designed four guide RNAs VvWRKY52 transcription factor gene with system, and obtained transgenic plants via Agrobacterium-mediated transformation, somatic embryos Thompson Seedless cultivar. Analysis first-generation verified 22 mutant 72 T-DNA-inserted plants. Of these, 15 lines carried seven were heterozygous. A range RNA-guided events, including large deletions, found plants, while smaller deletions comprised majority detected mutations. Sequencing potential off-target sites all targets revealed no events. addition, knockout increased resistance Botrytis cinerea. We conclude that allows precise represents useful functional analysis molecular breeding.