Fig. 2

Homozygous targeted mutagenesis of GmLHY1a/1b/2a/2b induced by CRISPR/Cas9. a Self-crossed scheme for generating a homozygous mutant that contains no Cas9 vector. (+/+) indicates that Cas9 was homozygous. (+/−) indicates that Cas9 was heterozygous. (−/−) indicates that Cas9 was deleted; b PCR-based genotyping results for the Cas9 gene in the T1 generation. (+) indicates that the gene was detected, (−) indicates that the gene was not detected; c Detailed sequence of the target site GmLHY2b in the T1–15 line; d Detailed sequence of the target site GmLHY2a in the T1–15 line; e Detailed sequence of the target site GmLHY1a in the T1–15 line; f Detailed sequence of the target site GmLHY1b in the T1–15 line. ‘-’ represents the number of deleted nucleotides. The red frames indicate the location of the targets; g Multiple alignment of the amino acid sequences of the quadruple mutant of GmLHY