Identification of CRISPR/Cas9 knockout targets and tissue expression analysis of circadian clock genes GmLNK1/2 , GmRVE4/8 , and GmTOC1 in soybean

Circadian clock genes play an important role in improving crop yield. LNK1 , LNK2 , RVE4 , RVE8 , and TOC1 are important circadian clock genes in plants. Homologous genes of AtLNK1 , AtLNK2 , AtRVE4 , AtRVE8 , and AtTOC1 in soybean were found by evolutionary tree analysis. These genes were expressed in soybean roots, stems and leaves. The knockout vectors of these genes were successfully constructed by using CRISPR/Cas9 gene editing technology. Transformation system of soybean root hair and RT-PCR were used to identify 13 genes targets effectively. The results of this study provide important target informations for further obtaining soybean mutant materials, and a foundation for further studying the function of circadian clock genes.