A Fe(Ⅱ)- and 2-oxoglutarate-dependent dioxygenase,named as GmF6′H1,was cloned from soybean variety ‘Heinong 35’.Expression of GmF6′H1 in different organs was examined by Quantitative(Q)-PCR analysis.The expression of GmF6′H1 was the highest in the roots,followed by in the pods,leaves and stems.Its expression showed weakly response to 2,4-D treatment and increased in a time-dependent manner after salicylic acid and kinetin treatment separately.With the time of treatment prelonged,GmF6′H1 expression levels steadily declined,and finally close to the level before the treatments.Purified protein of GmF6′H1 was able to catalyze feruloyl coenzyme A to scopoletin.By Agrobacterium tumefaciens mediated transformation,GmF6′H1 were transformed into the mutant Atf6′h1 of Arabidopsis thaliana.Compared with mutant plants,the external phenotype of the transgenic plants had no obvious difference,while its coumarin content in the roots increased the level close to the wild type of A.thaliana.