Mutation of GmIPK1 gene using CRISPR/Cas9 reduced phytic acid content in soybean seeds

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Title
Mutation of GmIPK1 gene using CRISPR/Cas9 reduced phytic acid content in soybean seeds
Author(s)
J H Song; Gilok Shin; H J Kim; Saet Buyl Lee; Ju Yeon Moon; Jae Cheol Jeong; H K Choi; I A Kim; H J Song; Cha Young Kim; Y S Chung
Bibliographic Citation
International Journal of Molecular Sciences, vol. 23, no. 18, pp. 10583-10583
Publication Year
2022
Abstract
Phytic acid (PA) acts as an antinutrient substance in cereal grains, disturbing the bioavailability of micronutrients, such as iron and zinc, in humans, causing malnutrition. GmIPK1 encodes the inositol 1,3,4,5,6-pentakisphosphate 2-kinase enzyme, which converts myo-inopsitol-1,3,4,5,6-pentakisphosphate (IP5) to myo-inositol-1,2,3,4,5,6-hexakisphosphate (IP6) in soybean (Glycine max L.). In this study, for developing soybean with low PA levels, we attempted to edit the GmIPK1 gene using the CRISPR/Cas9 system to introduce mutations into the GmIPK1 gene with guide RNAs in soybean (cv. Kwangankong). The GmIPK1 gene was disrupted using the CRISPR/Cas9 system, with sgRNA-1 and sgRNA-4 targeting the second and third exon, respectively. Several soybean Gmipk1 gene-edited lines were obtained in the T0 generation at editing frequencies of 0.1?84.3%. Sequencing analysis revealed various indel patterns with the deletion of 1?9 nucleotides and insertions of 1 nucleotide in several soybean lines (T0). Finally, we confirmed two sgRNA-4 Gmipk1 gene-edited homozygote soybean T1 plants (line #21-2: 5 bp deletion; line #21-3: 1 bp insertion) by PPT leaf coating assay and PCR analysis. Analysis of soybean Gmipk1 gene-edited lines indicated a reduction in PA content in soybean T2 seeds but did not show any defects in plant growth and seed development.
Keyword
SoybeanPhytic acidGenome editingCRISPR/Cas9sgRNAGmIPK1
ISSN
1661-6596
Publisher
MDPI
DOI
http://dx.doi.org/10.3390/ijms231810583
Type
Article
Appears in Collections:
Jeonbuk Branch Institute > Biological Resource Center > 1. Journal Articles
Jeonbuk Branch Institute > 1. Journal Articles
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