Abstract: FLOWERING LOCUS C（FLC）, a MADS box transcription factor, is the key repressor of vernalization in plants flowering pathway. Soybean is a non-vernalized crop, and the function of soybean FLC needs to be further studied. To investigate the function of soybean FLC in flowering regulation, we used bioinformatics methods to analyze the homology alignment, protein structure and tissue-specific expression of soybean FLC gene, and also verified the flowering time of different haplotypes of soybean FLC. The results showed that two soybean FLC-like genes were identified in 128 MADS-box gene, GmFLC-like 1（Glyma.05 G148700） and the newly discovered GmFLC-like 2（Glyma.08 G105400）, both of them had a highly conserved MADS-box domain, a relatively conserved K-box domain and a non-conserved C-terminal domain. The expression level of GmFLC-like 1 was higher in leaves, flower and pod, while the expression level of GmFLC-like 2 was higher in flower, pod, root and nodule. It was found that after single base mutations in the exon of GmFLC-like 1 and the intron of GmFLC-like 1 or GmFLC-like 2, the flowering time was significantly delayed, indicating that GmFLC-like 1 and GmFLC-like 2 may be involved in the regulation of soybean flowering time.