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【Objective】The present paper aimed to study whether PpCASLUU3 was involved in the hypersensitive reaction in Physcomitrella patens after infection with B.cinerea. Knockout vector PpCASLUU3-PTN182 was constructed, which could provide theoretical basis for plant defense of landing plant. 【Method】In the principle, genome DNA extracted from P. patens was used as a template. With primers designed using Primer Premier 5, the homologous upstream and downstream fragments were amplified, and cloned into pMD-19 T. After being sequenced, the upstream and downstream genes were obtained with restriction enzyme EcoR I and EcoR V, BamH I and Sma I, respectively. Meanwhile PTN182 was digested with the same enzyme. Used T4-DNA ligase, the recombinant vector was transferred into E.coli DH5α. Positive clones were screened out, and the downstream arm was connected to PpCASLUU3.1-PTN182 vector by the same method.【Result】Complete PpCASLUU3.1-PTN182-ppcasluu3.2 knockout vector was obtained by PCR detection and enzyme digestion verification. After sequencing analysis, the similarity of inserted sequences PpCASLUU3.1 and PpCASLUU3.2 were 100 % and 99.63 % respectively.【Conclusion】It indicated that the knockout vector PpCASLUU3-PTN182 was successfully constructed. [ABSTRACT FROM AUTHOR]