DWF4 and CPD are key brassinosteroids (BRs) biosynthesis enzyme genes. To explore the function of Populus euphratica DWF4 (PeDWF4) and CPD (PeCPD), Arabidopsis thaliana transgenic lines (TLs) expressing PeDWF4, PeCPD or PeDWF4 plus PeCPD, namely PeDWF4-TL, PeCPD-TL and PeCP/DW-TL, were characterized. Compared with wild type (WT), the changes of both PeDWF4-TL and PeCPD-TL in plant heights, silique and hypocotyls lengths and seed yields were similar, but in bolting time and stem diameters, they were opposite. PeCP/DW-TL was more in plant heights and the lengths of primary root, silique, and fruit stalk, but less in silique numbers and seed yields than either PeDWF4-TL or PeCPD-TL. PeDWF4 and PeCPD specially expressed in PeDWF4-TL or PeCPDTL, and the transcription level of PeDWF4 was higher than that of PeCPD. In PeCP/DW-TL, their expressions were all relatively reduced. Additionally, the expression of PeDWF4 and PeCPD differentially made the expression levels of AtDWF4, AtCPD, AtBR6OX2, AtFLC, AtTCP1 and AtGA5 change in the TLs. The total BRs contents were PeDWF4-TL greater than PeCP/DW-TL greater than WT greater than PeCPD-TL. These results imply that PeDWF4 is functionally not exactly the same as PeCPD and there may be a synergistic and antagonistic effects in physiology between both of them in the regulation of plant growth and development.