In this experiment,the 1 426 bp promoter fragment of LcCu/Zn-SOD3 from embryogenic callus of the ancient litchi tree ‘Songli’ which named ProLcCSD3(GenBank:KF672186.1) was cloned by means of adapter chromosome walking method.The bioinformatics analysis showed that ProLcCSD3 contains a number of environmental stress response elements,hormone response elements,endosperm expression elements and also might be regulation by MYB and WRKY transcription factors.A novel plant expression vector p1301-proLcCSD3-GUS was constructed with ProLcCSD3 replace 35S promoter in the pCAMBIA1301 vector by using the double-digested method and then transferred into Agrobacterium strain EHA105 and GV3101.Furthermore,transformation analysis showed that ProLcCSD3 has promoter activity by the infection of tobacco leaves and could drive the downstream reporter GUS gene expression in the roots,stems and leaves by Agrobacterium-mediated transformation of Arabidopsis.And stress treatments showed that ProLcCSD3 could response to some abiotic stresses,such as NaCl,PEG-6000,ABA,MeJA and wounding.Our results demonstrated that LcCu/Zn-SOD3 might response to abiotic stresses and participate in the signal transduction pathways of hormones in the ancient litchi tree.