Abstract: As one of the three fast-growing tree species in the world,Eucalyptus has high value in economic,ecological,medicinal and other aspects. Due to the high genetic heterozygosity of Eucalyptus,many majoreconomic traits might be jointly regulated by multiple genes,and conventional gene editing methods could notmeet the requirements for efficient screening of Eucalyptus target gene editing and transformation. Usingm Cherry fluorescent protein as a selection marker might reduce the identification workload after transformation.In this study,a CRISPR/Cas9 vector containing the 35 S promoter to promote the m Cherry marker gene wasconstructed using Eucalyptus urophylla as the material to perform efficient visual editing of the Eucalyptus gene.The m Cherry fluorescent protein was used as the selection marker to screen positively transformed progenies,and the adventitious bud genome containing fluorescent markers were extracted for PCR identification andanalysis. The results showed that the editing vector PHEE401-35 S-m Cherry was successfully constructed. After the callus of Eucalyptus urophylla transformed,there was obvious red fluorescence under the light of 580 nm,and the target fragment with the same size as 35 S-m Cherry band was amplified by PCR identification. The studyprovided a visual screening method for the Eucalyptus gene editing.