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Mutation of GmIPK1 Gene Using CRISPR/Cas9 Reduced Phytic Acid Content in Soybean Seeds.


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.

SUBMITTER: Song JH 

PROVIDER: S-EPMC9504718 | biostudies-literature | 2022 Sep

REPOSITORIES: biostudies-literature

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Mutation of <i>GmIPK1</i> Gene Using CRISPR/Cas9 Reduced Phytic Acid Content in Soybean Seeds.

Song Ji Hyeon JH   Shin Gilok G   Kim Hye Jeong HJ   Lee Saet Buyl SB   Moon Ju Yeon JY   Jeong Jae Cheol JC   Choi Hong-Kyu HK   Kim In Ah IA   Song Hyeon Jin HJ   Kim Cha Young CY   Chung Young-Soo YS  

International journal of molecular sciences 20220913 18


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. <i>GmIPK1</i> encodes the inositol 1,3,4,5,6-penta<i>kis</i>phosphate 2-kinase enzyme, which converts <i>myo</i>-inopsitol-1,3,4,5,6-penta<i>kis</i>phosphate (IP<sub>5</sub>) to <i>myo</i>-inositol-1,2,3,4,5,6-hexakisphosphate (IP<sub>6</sub>) in soybean (<i>Glycine max</i> L.). In this study, for developing soybean with lo  ...[more]

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