Molecular characterization and expression analysis of chalcone synthase gene during flower development in tree peony ( Paeonia suffruticosa )

Abstract

Chalcone synthase (CHS, EC: 2.3.1.74) is a key enzyme in the flavonoid and anthocyanin biosynthesis pathway. In order to investigate the role of CHS in tree peony flower coloration mechanism, we isolated and characterized the CHS gene from Paeonia suffruticosa cv. Yu Ji Yan Zhuang and analyzed its spatial and temporal expression patterns during floral development. The cDNA sequence of the CHS gene in P. suffruticosa (Ps-CHS 1, genbank accession no. GQ483511) was 1475 bp in full length containing an opening reading frame (ORF) of 1185 bp that encoded a 394 amino acid polypeptide. Bioinformatic analysis showed that, Ps-CHS1 possessed all the conserved active sites for the CHS function as well as the family signature. Sequence alignment and phylogenetic analysis revealed that Ps-CHS1 shared high homology with CHS from plants in Salicaceae, Malvaceae and Rosaceae. The homology-based structural modeling showed that Ps-CHS1 had the typical structure of CHS. Southern blot analysis indicated that CHS was encoded by a small multigene family in the genome of tree peony. Anthocyanidin content in full-opening flower petals accumulated to the highest level. Real-time polymerase chain reaction amplification (PCR) analysis indicated that, Ps-CHS 1 showed the highest transcript abundance in petals, moderate levels in sepals, low levels in leaves and stamens, and the lowest levels in carpels. Ps-CHS 1 was actively expressed during flower development and increased gradually until reached maximal expression when flower fully opened. These results indicated that PsCHS 1 was involved in the flower pigmentation of tree peony.