The invention relates to application of a soybean bHLH transcription factor GmPIF1 gene in promoting isoflavone synthesis, and belongs to the field of gene engineering and molecular biology. Accordingto the application of the soybean bHLH transcription factor GmPIF1 gene in promoting isoflavone synthesis, the nucleotide sequence of the soybean bHLH transcription factor GmPIF1 gene is shown as SEQID No. 1. The application has the beneficial effects as follows: the GmPIF1 promotes accumulation of isoflavone in soybean hairy roots, a new method is provided for increasing the content of soybeanisoflavone, the soybean isoflavone can be synthesized more efficiently through the bioengineering technology and the gene regulation and control method, and the defects that the artificial cultivationbreeding period is long, hybridization traits are unstable, and screening is difficult are overcome. The transcription factor GmPIF1 gene is transferred into soybean hairy roots, so that the transcription factor GmPIF1 gene is overexpressed in the hairy roots, the content of isoflavone in the hairy roots is increased, and the expression of related enzyme genes in an isoflavone synthetic pathway is regulated.