Multiplex genome editing targeting soybean with ultra-low anti-nutritive oligosaccharides
Wenxin Lin,
Huaqin Kuang,
Mengyan Bai,
Xiaomeng Jiang,
Pengfei Zhou,
Yinghua Li,
Bo Chen,
Huarong Li,
Yuefeng Guan
Affiliations
Wenxin Lin
College of Resources and Environment, Fujian Agriculture and Forestry University, Fuzhou 350002, Fujian, China
Huaqin Kuang
Guangdong Provincial Key Laboratory of Plant Adaptation and Molecular Design, Guangzhou Key Laboratory of Crop Gene Editing, Innovative Center of Molecular Genetics and Evolution, School of Life Sciences, Guangzhou University, Guangzhou 510006, Guangdong, China; FAFU-UCR Joint Center for Horticultural Biology and Metabolomics, Fujian Agriculture and Forestry University, Fuzhou 350002, Fujian, China
Mengyan Bai
College of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou 350002, Fujian, China
Xiaomeng Jiang
College of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou 350002, Fujian, China
Pengfei Zhou
Bioinformatics Department, School of Life Sciences and Technology, Tongji University, Shanghai 200092, China
Yinghua Li
KingAgroot CropScience Co., Ltd., Qingdao 266500, Shandong, China
Bo Chen
KingAgroot CropScience Co., Ltd., Qingdao 266500, Shandong, China
Huarong Li
KingAgroot CropScience Co., Ltd., Qingdao 266500, Shandong, China
Yuefeng Guan
Guangdong Provincial Key Laboratory of Plant Adaptation and Molecular Design, Guangzhou Key Laboratory of Crop Gene Editing, Innovative Center of Molecular Genetics and Evolution, School of Life Sciences, Guangzhou University, Guangzhou 510006, Guangdong, China; FAFU-UCR Joint Center for Horticultural Biology and Metabolomics, Fujian Agriculture and Forestry University, Fuzhou 350002, Fujian, China; Corresponding author.
Soybean is the primary source of plant protein for humans. Owing to the indigestibility of the raffinose family of oligosaccharides (RFO), raffinose and stachyose are considered anti-nutritive factors in soybean seeds. Low-RFO soybean cultivars are generated by mutagenesis of RFO biosynthesis genes, but the carbohydrate profiles invite further modification to lower RFOs. This study employed a pooled multiplex genome editing approach to target four seed-specifically expressed genes mediating RFO biosynthesis, encoding three raffinose synthases (RS2, RS3, and RS4) and one stachyose synthase. In T1 progeny, rs2/rs3 and rs4/sts homozygous double mutants and a rs2/rs3/rs4/sts quadruple mutant (rfo-4m) were characterized. The rs2/rs3 mutant showed reduced raffinose and stachyose contents, but the rs4/sts mutant showed only reduced stachyose in seeds. The RFO contents in the rfo-4m mutant were almost eliminated. Metabolomic analysis showed that the mutation of four RFO biosynthesis genes led to a shift of metabolic profile in the seeds, including the accumulation of several oligosaccharides-related metabolites. These mutants could contribute to precision breeding of soybean cultivars for soy food production.
