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Target-specific mutations efficiency at multiple loci of CRISPR/Cas9 system using one sgRNA in soybean

文献类型： 外文期刊

作者： Zhang, Ling 1 ; Wang, Yingzhe 1 ; Li, Tong 1 ; Qiu, Hongmei 1 ; Xia, Zhengjun 2 ; Dong, Yingshan 1 ;

作者机构： 1.Jilin Acad Agr Sci, Jilin Prov Key Lab Agr Biotechnol, Changchun 130033, Jilin, Peoples R China

2.Chinese Acad Sci, Innovat Acad Seed Design, Northeast Inst Geog & Agroecol, Harbin, Peoples R China

关键词： Soybean; CRISPR; Cas9; GmPDS genes; Mutation efficiency

期刊名称：TRANSGENIC RESEARCH （ 影响因子：2.788； 五年影响因子：2.377 ）

ISSN： 0962-8819

年卷期： 2021 年 30 卷 1 期

页码：

收录情况： SCI

摘要： Soybean has a palaeopolyploid genome with nearly 75% of the genes present in multiple copies. Although the CRISPR/Cas9 system has been employed in soybean to generate site-directed mutagenesis, a systematical assessment of mutation efficiency of the CRISPR/Cas9 system for the multiple-copy genes is still urgently needed. Here, we successfully optimize one sgRNA CRISPR/Cas9 system in soybean by testing the efficiency, pattern, specificity of the mutations at multiple loci of GmFAD2 and GmALS. The results showed that simultaneous site-directed mutagenesis of two homoeologous loci by one sgRNA, the mutation frequency in the T0 generation were 64.71% for GmPDS, 60.0% for GmFAD2 and 42.86% for GmALS, respectively. The chimeric and heterozygous mutations were dominant types. Moreover, association of phenotypes with mutation pattern at target loci of GmPDS11 and GmPDS18 could help us further demonstrate that the CRISPR/Cas9 system can efficiently generate target specific mutations at multiple loci using one sgRNA in soybean, albeit with a relatively low transformation efficiency.

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