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Research paper | Open Access

Multiplex genome editing targeting soybean with ultra-low anti-nutritive oligosaccharides

Wenxin Lina,1, Huaqin Kuangb,c,1, Mengyan Baid, Xiaomeng Jiangd, Pengfei Zhoue, Yinghua Lif, Bo Chenf, Huarong Lif, Yuefeng Guanb,c( )
College of Resources and Environment, Fujian Agriculture and Forestry University, Fuzhou 350002, Fujian, China
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
College of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou 350002, Fujian, China
Bioinformatics Department, School of Life Sciences and Technology, Tongji University, Shanghai 200092, China
KingAgroot CropScience Co., Ltd., Qingdao 266500, Shandong, China

1 These authors contributed equally to this work.

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Abstract

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.

Keywords

Genome editingSoybeanStachyoseRaffinose family of oligosaccharidesRaffinosePrecision breeding

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The Crop Journal
Volume 11 Issue 3,
June 2023
Pages 825-831
DOI: 10.1016/j.cj.2023.01.001
Cite this article:
Lin W, Kuang H, Bai M, et al. Multiplex genome editing targeting soybean with ultra-low anti-nutritive oligosaccharides. The Crop Journal, 2023, 11(3): 825-831. https://doi.org/10.1016/j.cj.2023.01.001

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Received: 12 September 2022
Revised: 26 November 2022
Accepted: 02 January 2023
Published: 21 January 2023
© 2022 Crop Science Society of China and Institute of Crop Science, CAAS.

This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
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