In order to study the role of PfNLR gene family in Paulownia fortunei resistance to Paulownia witches'broom(PaWB),bioinformatics methods were used to comprehensively analyze the composition,physico-chemical properties,chromosomal localization,evolution,structure,cis-acting elements and tissue expression spe-cificity of P.fortunei NLR(PfNLR)family members.Combined with the miRNA and transcriptome data before and after phytoplasma infecting Paulownia,the genes associated with resistance to PaWB were screened.The res-ults showed that 199 PfNLRs in P.fortunei belonged to 6 subfamilies;91.9%of PfNLRs were located on 18 chro-mosomes of P.fortunei,of which 62.8%were clustered,among them,there were 10 chromosome segment re-peats and 1 chromosome tandem repeats in 23 PfNLRs;PfNLR contains hormonal and stress elements;integrated transcriptome and miRNA analysis found that there were 156 pairs of targeted relationships between PfNLRs and miRNAs;most of PfNLRs were expressed in buds and roots;of which,16 PfNLRs were significantly different be-fore and after phytoplasma infection.Further analysis revealed that PfNLR181 mediated by pf-miRNA482 played a role in resistance of PaWB.These results could provide a theoretical basis for analyzing the function of PfNLR protein.