TY  - JOUR
AU  - Wang, Jie
AU  - Kuang, Huaqin
AU  - Zhang, Zhihui
AU  - Yang, Yongqing
AU  - Yan, Long
AU  - Zhang, Mengchen
AU  - Song, Shikui
AU  - Guan, Yuefeng
PY  - 2020
TI  - Generation of seed lipoxygenase-free soybean using CRISPR-Cas9
JO  - The Crop Journal
SN  - 2095-5421
SP  - 432
EP  - 439
VL  - 8
IS  - 3
AB  - 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 T0 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 T1 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 T2 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.
UR  - https://doi.org/10.1016/j.cj.2019.08.008
DO  - 10.1016/j.cj.2019.08.008