In order to verify the mutation efficiency and identify the mutation type of different targets,two pairs of targets were designed in the first and second exons of GmFAD2-1(g3)and GmFAD-2b(g6)genes respectively,and their activity was verified by in vitro activity test.At the same time,CRISPR/Cas9 vectors with different targets were constructed,and the mutation efficiency and type were further verified by Agrobacterium rhizogenes transfor-mation method.The results showed that the activity of the target designed at the first exon of the gene was signifi-cantly higher than that designed at other locations.The content of GC in the designed target was 52.6％,36.8％,31.5％and 21.1％respectively,indicating that the higher the content of GC,the stronger the activity of the target.The efficiency of g3-Target l with U6 as promoter was 92％,and the mutation types were mainly base insertion,deletion and substitution;the mutation efficiency of g3-Target2 was 36％,and the main mutation types were inser-tion and deletion.The mutation efficiency of g6-Targetl with U6 as promoter was 84％,and the main mutation types were insertion,deletion and substitution;the mutation efficiency of g6-Target2 was 8％,and the main muta-tion types were insertion and deletion of base.
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