Fig. 1

Maps of pYLgRNA and pYLCRISPR/Cas9-DB vectors"

Fig. 2

Positions of three targets in the GmFAD2-1A gene locus The white boxes on the left and the right represent 5' and 3' noncoding regions; the black box stands for CDS; the solid line represents the intron."

Table 1

Primer sequence"

Fig. 3

Schematic diagram of the pYLCRISPR/Cas9-FAD2-1A-gRNA vector construction"

Table 2

Mediums and compositions in soybean transformation"

Fig. 4

Identification of recombinant plasmids A: Electrophoresis of PCR detection of clony; M: DNA marker (DM2000); 1: H2O blank control; 2: pYLCRISPR/Cas9-DB plasmid; 3-7: FAD2-1A gene knockout positive single clones. B: Identification of the pYLCRISPR/Cas9-FAD2-1A-gRNA plasmid digested with Asc I; M: 1 kb DNA ladder marker; 1: pYLCRISPR/Cas9- FAD2-1A-gRNA."

Fig. 5

Conversion process and the results of leaf painting A: transformation soybean cotyledonine by agrobacterium; 1: germination of sterile seedlings; 2: co-cultivation; 3: shoot induction; 4: shoot elongation; 5: rooting of the resistant shoot; 6: seedling acclimatization. B: identification of the herbicide resistance of the leaves of T0 transgenic soybean plants using glufosinate; a: positive transgenic plant; b: negative transgenic plant."

Fig. 6

Identification of GmFAD2-1A mutant A: PCR detection of T1 transgenic soybean; M: DNA marker (DM2000); 1: H2O blank control; 2: WT H3; 3-7: positive seedling of glufosinate tolerence of transgenic soybean. B: sequence alignment of GmFAD2-1A mutant compared to the WT line; C: sequence alignment of GmFAD2-1A mutant and wild-type protein."

Table 3

Agronomic traits of the transgenic soybean"

Fig. 7

GmFAD2-1A mutants phenotype and the fatty acids content OA: oleic acid; LA: linoleic acid; ALA: linolenic acid; PA: palmitic acid; SA: stearic acid."