Abstract: In order to explore the characteristics of chloroplast genome of Rubus alceifolius and its phylogenetic relationship with chloroplast genome of the same family, the Illumina HiSeq 4000 platform was used to sequence, assemble and annotate Rubus alceifolius, and its sequence characteristics, codon preference, sequence repeats, simple sequence repeats（SSR） and phylogeny were analyzed. The results showed that the chloroplast genome of Rubus alceifolius showed a typical tetrad structure, with a total length of 156 266 bp, including a large single copy region of 85 874 bp, a small single copy region of 18 850 bp and a reverse complementary repeat region of 25 771 bp. A total of 128 genes were detected, including 86 protein coding genes, 34 tRNA genes and 8 rRNA genes. Codon preference analysis showed that leucine（Leu） had the largest number of codons（5 189）, while cysteine（Cys） had the least number of codons（590）. A total of 39 long repeats and 52 SSR loci were detected from the chloroplast genome of Rubus alceifolius. Phylogeny ana-lysis showed that Rubus reflexus var. lanceolobus, Rubus cochinchinensis, Rubus rufus and Rubus alceifolius were clustered into one branch, and the relationship was the closest. This study provides scientific basis for molecular markers, species identification, kinship analysis and genetic diversity analysis of Rosaceae and Rubus.