Abstract: In order to better understand the Paphiopedilum dianthum and develop its horticultural value, the transcriptome sequencing of flower buds P. dianthum was performed using RNA-seq technique. The results showed that a total of 95,659 Unigenes were obtained, and a total of 61,629 Unigene annotations were found, accounting for 64.43% of all Unigene annotations by comparing Unigene to NR, KOG, SwissProt, KEGG and other databases. The number of unigene annotated genes in CDD, KOG, NR, NT, PFAM, SWISSPROT, TREMBL, GO and KEGG databases were 33,589, 28,405 and 45,568, 56,635, 23,870, 52,141, 44,973, 54,934, 4,893 respectively. The annotated results showed that the most homologous sequences of P. dianthum were Elaeis guineensis. It could be divided into 71 groups in 3 broad categories in the GO annotation, and 25 functional categories in the KOG database. According to KEGG database and results of pathway enrichment, a total of 4,893 Unigenes were involved in 327 metabolic pathways of 23 classes. The SSR detection of Unigene by MISA software showed that 7,613 of 95,659 Unigenes had SSR, and a total of 8,160 SSR loci were found. The length of the SSR loci ranged from 10 to 230 bp, with an average length of 66.95 bp. Dinucleotide had the highest SSR richness, accounting for 33.72%, followed by mononucleotide and trinucleotide, accounting for 32.12% and 26.11%, respectively. In this study, a large number of gene sequences were obtained by transcriptome sequencing of P. dianthum, and the general expression of genes in the organs of P. dianthum was understood, which provided references for the exploration and utilization of genes related to organ development of P. dianthum, the development of SSR molecular markers, and the sequencing and assembly of its genome. It also laid a foundation for further research on the molecular biology of P. dianthum.