title: Rapid generation of genetic diversity by multiplex CRISPR/Cas9 genome editing in rice.
abstract: The clustered regularly interspaced short palindromic repeats (CRISPR)-associated endonuclease 9 (CRISPR/Cas9) system has emerged as a promising technology for specific genome editing in many species. Here we constructed one vector targeting eight agronomic genes in rice using the CRISPR/Cas9 multiplex genome editing system. By subsequent genetic transformation and DNA sequencing, we found that the eight target genes have high mutation efficiencies in the T0generation. Both heterozygous and homozygous mutations of all editing genes were obtained in T0plants. In addition, homozygous sextuple, septuple, and octuple mutants were identified. As the abundant genotypes in T0transgenic plants, various phenotypes related to the editing genes were observed. The findings demonstrate the potential of the CRISPR/Cas9 system for rapid introduction of genetic diversity during crop breeding.
authors.affiliation: State Key Laboratory of Rice Biology, China National Rice Research Institute, Chinese Academy of Agricultural Sciences, Hangzhou, 310006, China.|Key Laboratory of Plant Functional Genomics, Ministry of Education, Yangzhou University, Yangzhou, 225009, China.
authors.forename: Lan
authors.initials: L
authors.lastname: Shen
authors.affiliation: State Key Laboratory of Rice Biology, China National Rice Research Institute, Chinese Academy of Agricultural Sciences, Hangzhou, 310006, China.
authors.forename: Yufeng
authors.initials: Y
authors.lastname: Hua
authors.affiliation: State Key Laboratory of Rice Biology, China National Rice Research Institute, Chinese Academy of Agricultural Sciences, Hangzhou, 310006, China.
authors.forename: Yaping
authors.initials: Y
authors.lastname: Fu
authors.affiliation: Lianyungang Academy of Agricultural Sciences, Lianyungang, 222006, China.
authors.forename: Jian
authors.initials: J
authors.lastname: Li
authors.affiliation: State Key Laboratory of Rice Biology, China National Rice Research Institute, Chinese Academy of Agricultural Sciences, Hangzhou, 310006, China.
authors.forename: Qing
authors.initials: Q
authors.lastname: Liu
authors.affiliation: State Key Laboratory of Rice Biology, China National Rice Research Institute, Chinese Academy of Agricultural Sciences, Hangzhou, 310006, China.
authors.forename: Xiaozhen
authors.initials: X
authors.lastname: Jiao
authors.affiliation: College of Life Sciences, Shanxi Agricultural University, Taigu, 030801, China.
authors.forename: Gaowei
authors.initials: G
authors.lastname: Xin
authors.affiliation: State Key Laboratory of Rice Biology, China National Rice Research Institute, Chinese Academy of Agricultural Sciences, Hangzhou, 310006, China.
authors.forename: Junjie
authors.initials: J
authors.lastname: Wang
authors.affiliation: College of Life Sciences, Shanxi Agricultural University, Taigu, 030801, China.
authors.forename: Xingchun
authors.initials: X
authors.lastname: Wang
authors.affiliation: Key Laboratory of Plant Functional Genomics, Ministry of Education, Yangzhou University, Yangzhou, 225009, China. cjyan@yzu.edu.cn.
authors.forename: Changjie
authors.initials: C
authors.lastname: Yan
authors.affiliation: State Key Laboratory of Rice Biology, China National Rice Research Institute, Chinese Academy of Agricultural Sciences, Hangzhou, 310006, China. wangkejian@caas.cn.
authors.forename: Kejian
authors.initials: K
authors.lastname: Wang
mesh_terms: D001483:Base Sequence; D064113:CRISPR-Cas Systems; D000072669:Gene Editing / Q000379:methods*; D017343:Genes, Plant / Q000235:genetics; D014644:Genetic Variation; D018745:Genome, Plant / Q000235:genetics*; D005838:Genotype; D009154:Mutation; D012275:Oryza / Q000235:genetics*; D010641:Phenotype; D000069600:Plant Breeding / Q000379:methods; D030821:Plants, Genetically Modified
keywords: CRISPR/Cas9; agronomic genes; genome editing; rice