Abstract: 【Objective】To explore the transcriptional regulation mechanism on rice allelopathy resistant weeds，and provide theoretical basis for breeding rice of resistant weeds.It provided basis for the genetic modification and improvement of rice allelopathy traits，and also provided a new way for the prevention and control of weeds.【Method】Using SMART technology，cDNA library of rice under barnyard grass stress was constructed in Y2H by homologous recombination. The decoy vector pGBK-OsMYB57 with C-terminal deletion of OsMYB57 was constructed using restriction endonuclease BamHⅠ and NdeⅠ，and its self-activation activity was detected according to Yeastmaker^TMYeast Transformation System 2. The interaction protein of OsMYB57 was screened by yeast two-hybrid technology，and pGAD vector was constructed with the interaction protein coding sequence，and the interaction protein of Y2HGold strain was co-transformed with bait vector. The interaction between proteins was further verified by BiFC technology.【Result】The size of yeast cDNA library was 1.36×10⁷ CFU/mL，the inserted fragment was 250-2000 bp，the average length was more than 1000 bp，and the recombinant rate was 100%，showed that the library was large and of good quality. Four annotated interacting proteins，RZFP34 16，4HTR 50，BM1PD 51 and GTP1 74，were screened out. RZFP34，4HTR，BMIPD and GTP1 in-teracted with OsMYB57 in plant cells. The RZFP34 was an E3 subtype ubiquitin ligase that responded to abiotic stress in plants，and the promoter region of rice OsRZFP34 contained elements related to hormone response such as salicylic acid， jasmonic acid and MYB transcription factor binding domain related to allelopathy regulation of rice.【Conclusion】RZFP34 interacts with OsMYB57 and participates in allelopathy transcriptional regulation of rice.