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Title:: Generation ofgmnarkMutant with SupernodulationviaCRISPR-Cas9 in Soybean

Author(s):: BAI Meng-yan¹; 2; YUAN Jue-hui¹; 2; SUN Jia-feng¹; 2; LI Su-ning²; 3; GUAN Yue-feng²; (1.College of Resources and Environment, Fujian Agriculture and Forestry University, Fuzhou 350002, China; 2.FAFU-UCR Joint Center for Horticultural Biology and Metabolomics, Haixia Institute of Science and Technology, Fujian Agriculture and Forestry University, Fuzhou 350002, China; 3.College of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou 350002, China)

Keywords:: Soybean; Genome editing; Autoregulation of nodulation; Frameshift mutation

PACS:: -

DOI:: 10.11861/j.issn.1000-9841.2019.04.0525

Abstract:: In order to study the mechanism of the autoregulation of nodulation (AON) in the legumes, the gene editing technology of CRISPR-Cas9 system was used to create thegmnarkmutant of the soybean variety Huachun 6, and three specific sgRNAs were designed to locate the targeted geneGmNARKand constructed into a CRISPR-Cas9 knockout vector. Two sgRNAs with high editing efficiency, sgRNA-B and sgRNA-C, were selected for stable transformation through hairy root transformation experiments. Furthermore, eight mutants of different mutation types were screened in T₁generation and five of them had the phenotype of supernodulation which is verified by hydroponic experiment. In the further phenotypic analysis ofgmnark-Amutant, it was found thatgmnark-Aowned a larger number of nodules, smaller shoot and root and greener leaves than WT. These data proved that thegmnarkmutant created in the study is an important genetic material for studying the mechanism of AON pathway and the development of nodule. At the same time, this new germplasm resource has great potential in intercropping with other crops as ‘green manure’.

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Generation ofgmnarkMutant with SupernodulationviaCRISPR-Cas9 in Soybean(PDF)

《大豆科学》[ISSN:1000-9841/CN:23-1227/S]

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