Abstract: In order to enhance the efficiency of gene transformation via the pollen tube pathway，the timing of DNA inoculation，the concentration of DNA，and the different methods of DNA inoculation were studied in‘Qingxiang’walnut（Juglans regia L.）. The results showed that a period of 14 to 20 hours after pollination was the optimum time for the treatment of dripping exogenous DNA on the pistils. And a period of 24 hours was suitable for the treatment of injection. A DNA concentration of 350–500 μg · mL^-1 was optimal for gene transformation according to the fruit set rates and GUS analysis. The
fruits carrying the exogenous gene were obtained by using three methods of dripping DNA with or without the pistil cut and of the ovule injection，while the dripping DNA without the pistil cut treatment had the highest transformation rate（36.4%），followed by the dripping DNA with the pistil cut treatment
（20.7%）. The ovule injection treatment had the lowest fruit set rate（10.9%）due to mechanical damage during injection.

Key words: Juglans regia, exogenous DNA, pollen tube pathway, dripping on pistil, efficiency of transformation