Fig. 1

JN18 and MT72 fatty acid content gas chromatogramNote: A：JN18; B:MT72. The peak time from left to right is linolenic acid, oleic acid, linoleic acid, palmitic acid, stearic acid

Table 1

PCR Primer sequence

Fig. 2

GmFAD3 gene PCR amplification resultNote: M: DL10000 Marker; W: Negative control;1-3: JN18;4-6: MT72;A: GmFAD3A;B: GmFAD3B;C: GmFAD3C;D: GmFAD3C-1

Fig. 3

Multiple sequence alignment of sequencing resultsNote: A: Schematic diagram of mutation site;B: Amino acid multiple sequence alignment diagram

Fig. 4

Bioinformatics analysis result graphNote: A: protein conserved domain analysis;B: transmembrane structure analysis;C: hydrophilic and hydrophobic analysis;D: three-dimensional structure model prediction

Fig. 5

PCR result of recombinant plasmid bacterial solutionNote: M: DL10000Marker;W: Negative control;A: Extraction of total soybean RNA;B: Full-length cDNA sequence of GmFAD3C-1 gene;C: PCR detection of overexpression vector bacterial solution;D: PCR detection of editing vector bacteria;E: schematic diagram of editing vector construction

Fig. 6

PCR detection of some transgenic plantsNote: M: DL10000Marker; -: negative control;+: positive control, Figure;A: T1 generation 35s detection 1-3: JN18OET1 4-6: MT72OET1, Figure;B: T2 generation 35s detection 1-3: JN18OET2 4-6: MT72OET2, Figure;C: T1 generation Bar detection 1-3: JN18OET1 4-7: MT72OET1, Figure;D: T2 generation Bar detection 1-3: JN18OET2 4-6: MT72OET2, Figure;E: T1 generation Cas9 detection 1-2: JN18CRT1 3-5: MT72CRT1, Figure;F: T2 generation Cas9 detection 1-2: JN18CRT2 3-5: MT72CRT2

Fig. 7

Analysis of sgRNA mutation types in transgenic plantsNote: A: M: DL2000 Marker W: Negative control -: Untransformed plants +: Positive control 1-2: JN18CRT1 3-5: MT72CRT1 Figure;B: Sequence alignment of sgRNA mutation sites in some transgenic plants Figure;C: Partial sgRNA in transgenic plants Alignment of amino acid sequence of mutation site;D: Prediction of three-dimensional structure of some proteins

Fig. 8

Southern blotting detection of genetically modified soybeansNote: -: negative control;+: positive control; A: 1-6 JN18OET2;B: 1-6 MT72OET2;C:1-6 JN18CRT2;D:1-6 MT72CRT2

Fig. 9

Fluorescence quantitative PCR analysis of T2 transgenic positive plants

Fig. 10

Determination of enzyme activity in grains of T2 transgenic linesNote: A: standard curve; B: determination of enzyme activity in the seed of transgenic plant line; 1 enzyme unit (U) means the amount of enzyme which can transfor 1 μmol base in 1 min. U/mg refers to the number of U per mg of fatty acid dehydrogenase

Fig. 11

Determination of relative fatty acid content in grains of T2 generation transgenic lines