Background: Rose is one of most popular ornamental plants all over the world, it is of high economical value and great cultural importance. However, chilling damage restricts its application in cold area. To elucidate the metabolic response under low temperature stress, we conducted transcriptome and De novo analysis in Rosa xanthina f. spontanea.Results: A total of 124,106 unigenes from 9 databases were generated by De novo assembly, with mean length of 661bp under 4℃ and -20℃ stress (23℃ as control). Functional annotation and prediction of 55,084 unigenes were detected and 67.72% of these unigenes had significant similarity (Blast, E≤10-5) in the public databases. 468 DEGs were involved in three groups: biological process (64.84%), cellular component (9.38%) and molecular function (25.78%), KEGG analysis suggested that metabolic pathway, response to plant pathogen interaction, starch and sucrose metabolism, circadian rhythm plant and photosynthesis antenna proteins were significantly enriched under cold stress.Conclusions: Our study first reported response to cold stress at the transcriptome level in Rosa xanthina f. spontanea, which can provides a theoretical basis for further studies on the molecular mechanism of cold-resistance in rose. The results was shown that the expression trend of eight DEGs selected by qRT-PCR analysis at random was consistent with the results of high-throughput sequencing.