Mutagenesis of seed storage protein genes in Soybean using CRISPR/Cas9

Chenlong Li; Vi Nguyen; Jun Liu; Wenqun Fu; Chen Chen; Kangfu Yu; Yuhai Cui

doi:10.1186/s13104-019-4207-2

Mutagenesis of seed storage protein genes in Soybean using CRISPR/Cas9

BMC Res Notes. 2019 Mar 27;12(1):176. doi: 10.1186/s13104-019-4207-2.

Authors

Chenlong Li^{1

2

3}, Vi Nguyen⁴, Jun Liu^{4

5}, Wenqun Fu^{4

6}, Chen Chen^{4

7}, Kangfu Yu⁸, Yuhai Cui^{9

10}

Affiliations

¹ London Research and Development Center, Agriculture and Agri-Food Canada, London, ON, Canada. lichlong3@mail.sysu.edu.cn.
² Department of Biology, Western University, London, ON, Canada. lichlong3@mail.sysu.edu.cn.
³ State Key Laboratory of Biocontrol and Guangdong Key Laboratory of Plant Resource, School of Life Sciences, Sun Yat-sen University, Guangzhou, China. lichlong3@mail.sysu.edu.cn.
⁴ London Research and Development Center, Agriculture and Agri-Food Canada, London, ON, Canada.
⁵ Guangdong Academy of Agricultural Sciences, Guangzhou, China.
⁶ Department of Biological Science and Technology, Minnan Normal University, Zhangzhou, China.
⁷ Department of Biology, Western University, London, ON, Canada.
⁸ Agriculture and Agri-Food Canada, Harrow Research and Development Centre, Harrow, ON, Canada.
⁹ London Research and Development Center, Agriculture and Agri-Food Canada, London, ON, Canada. yuhai.cui@canada.ca.
¹⁰ Department of Biology, Western University, London, ON, Canada. yuhai.cui@canada.ca.

Abstract

Objective: Soybean seeds are an important source of vegetable proteins for both food and industry worldwide. Conglycinins (7S) and glycinins (11S), which are two major families of storage proteins encoded by a small family of genes, account for about 70% of total soy seed protein. Mutant alleles of these genes are often necessary in certain breeding programs, as the relative abundance of these protein subunits affect amino acid composition and soy food properties. In this study, we set out to test the efficiency of the CRISPR/Cas9 system in editing soybean storage protein genes using Agrobacterium rhizogenes-mediated hairy root transformation system.

Results: We designed and tested sgRNAs to target nine different major storage protein genes and detected DNA mutations in three storage protein genes in soybean hairy roots, at a ratio ranging from 3.8 to 43.7%. Our work provides a useful resource for future soybean breeders to engineer/develop varieties with mutations in seed storage proteins.

Keywords: CRISPR/Cas9; Genome editing; Seed storage proteins; Soybean.

MeSH terms

CRISPR-Cas Systems / genetics*
Genes, Plant / genetics*
Glycine max / genetics*
Mutation / genetics*
Plant Roots / genetics*
Plants, Genetically Modified / genetics*
Seed Storage Proteins / genetics*
Seeds / genetics*

Substances

Seed Storage Proteins

Grants and funding

A-base/Agriculture and Agri-Food Canada