Mosses have adapted to the Antarctic environment and are an ideal medium for studying plant resistance to abiotic stress. Chalcone synthase is the first committed enzyme in the flavonoid metabolic pathway, which plays an indispensable role in plant resistance to adversity. In this study, six genes (Pn021, PnCHS088, Pn270, PnCHS444, PnCHS768 and Pn847) were identified in the Antarctic moss Pohlia nutans Lindberg transcriptome by reverse transcription polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends (RACE). Sequence alignment and three-dimensional structure analysis revealed the conserved amino acid residues of the enzymes of the chalcone synthase family, including three catalytic residues (Cys164, His303 and Asn336) and two substrate recognition residues (Phe215 and Phe265). Phylogenetic analysis indicated that PnCHS088, PnCHS444 and PnCHS768 might be chalcone synthase but that Pn021 is more like stilbenecarboxylate synthase. These genes were located at the transition between fungi and advanced plants in the phylogenetic tree. In addition, real-time PCR analysis revealed that the expression profiles of the six P. nutans genes were influenced by diverse abiotic stresses as well as by abscisic acid and methyl jasmonate. The results presented here contribute to the study of the CHS gene family in polar mosses and further reveal the mechanisms underlying the adaptation of mosses to extreme environments.