Abstract: 【Objective】Explore the molecular mechanism of resistance to rust in Populus deltoides and provide the theoretical basis for the poplar molecular breeding.【Method】 In this study, eighty F1 progenies were selected for genetic analysis using P. detloides parents of ‘2-2'(female parent)and ‘2-38'(male parent)controlled pollination hybridization which possess substantial differences in resistance to rust. The F₁ progeny were sequenced by SLAF-seq, using the third generation genome of P. deltoides as reference. Therefore, specific DNA fragments were extracted to construct the SLAF-seq library. Meanwhile, a great amount of specific SNP sites were identified. Melampsora larici-populina(Kleb.)that caused foliar rust of poplar were identified by morphylogy and molecular tools.And then the rust infection levels were evaluated under the control condition through leaf disk method. The data were analyzed with Kruskal-Wallis test.【Result】Leaf disks were inoculated with the rust and hold for at least eight days. Phenotypes of uredinial numbers and uredinial size were analyzed with the help of stereoscopy. Totally, 8 723 SNP tags are linked to genetic map and 19 linkage groups with a total genetic distance of 3 610.67 cM were identified through linkage analysis of SNP markers. Thirty-seven closely linked loci(P <0.005)are revealed and locates in linkage groups I, IV, VI, XI, XV and XIX respectively with Kruskal-Wallis test.【Conclusion】Thirty-seven loci were developed by SLAF-seq and found to be tightly linked with the rust resistance in P. deltoides and one of the loci, Marker42056, is the nearest to resistance site genetically.