Generation of seed lipoxygenase-free soybean using CRISPR-Cas9

文献类型： 外文期刊

第一作者： Wang, Jie

作者： Wang, Jie;Zhang, Zhihui;Wang, Jie;Kuang, Huaqin;Zhang, Zhihui;Song, Shikui;Guan, Yuefeng;Yang, Yongqing;Yan, Long;Zhang, Mengchen

作者机构：

期刊名称：CROP JOURNAL （ 影响因子：4.407； 五年影响因子：5.687 ）

ISSN： 2095-5421

年卷期： 2020 年 8 卷 3 期

页码：

收录情况： SCI

摘要： Beany flavor induced by three lipoxygenases (LOXs, including LOX1, LOX2, and LOX3) restricts human consumption of soybean. It is desirable to generate lipoxygenase-free new mutant lines to improve the eating quality of soybean oil and protein products. In this study, a pooled clustered regularly interspaced short palindromic repeats (CRISPR)-CRISPR-associated protein 9 (Cas9) strategy targeting three GmLox genes (GmLox1, GmLox2, and GmLox3) was applied and 60 T-0 positive transgenic plants were generated, carrying combinations of sgRNAs and mutations. Among them, GmLox-28 and GmLox-60 were gmlox1gmlox2gmlox3 triple mutants and GmLox-40 was a gmlox1gmlox2 double mutant. Sequencing of T-1 mutant plants derived from GmLox-28, GmLox-60, and GmLox-40 showed that mutation in the GmLox gene was inherited by the next generation. Colorimetric assay revealed that plants carrying different combinations of mutations lost the corresponding lipoxygenase activities. Transgene-free mutants were obtained by screening the T-2 generation of lipoxygenase-free mutant lines (GmLox-28 and GmLox-60). These transgene- and lipoxygenase-free mutants could be used for soybean beany flavor reduction without restriction by regulatory frameworks governing transgenic organisms. (C) 2019 Crop Science Society of China and Institute of Crop Science, CAAS. Production and hosting by Elsevier B.V. on behalf of KeAiCommunications Co., Ltd.

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