Gene cloning and bioinformatics analysis of chalcone synthase gene from Daphniphyllum calycinum

doi:10.14088/j.cnki.issn0439-8114.2026.05.031

Abstract

Abstract: To reveal the characteristics and potential functions of the chalcone synthase gene (DcCHS) from Daphniphyllum calycinum, highly expressed candidate genes were screened based on the transcriptome data of Daphniphyllum calycinum, and after total RNA extraction and cDNA synthesis, the target genes were cloned using PCR technology. Bioinformatics methods were used to analyze the physicochemical properties, domains, subcellular localization and spatial structure of the genes, multiple sequence alignment and phylogenetic tree construction were performed, and the binding activity of CHS proteins with small molecules was verified by molecular docking. The results showed that the full lengths of the open reading frames (ORFs) of DcCHS1,DcCHS2 and DcCHS3 were 1 188, 1 188 and 1 023 bp, respectively, with G+C contents all greater than 51%, encoding 395, 395 and 340 amino acids, respectively.The molecular weights of three DcCHS proteins (DcCHS1, DcCHS2 and DcCHS3) were 43.08, 43.03 and 37.03 ku, respectively.All three DcCHS proteins were non-secretory proteins, among which DcCHS1 was a weakly acidic unstable protein, while DcCHS2 and DcCHS3 were weakly acidic stable proteins. Subcellular localization prediction showed that they were mainly located in the cytoplasm. The aliphatic indices of three DcCHS proteins were all greater than 90, and they all had no signal peptide or transmembrane domain. All three DcCHS proteins contained four conserved residues (Cys, Phe, His, Asn) of the CHS catalytic center and the CHS family characteristic sequence “WGVLFGFGPGLT”. Phylogenetic analysis showed that the CHS genes of Daphniphyllum calycinum were closely related to the CHS genes of citrus, Arabidopsis and other species. Molecular docking results showed that the DcCHS proteins had good binding ability with flavonoid small molecules such as naringenin and phloretin. The three DcCHS genes might play important roles in anthocyanin accumulation and abiotic stress response.

Key words: daphniphyllum calycinum, chalcone synthase (CHS), gene cloning, bioinformatics

CLC Number:

S567

CUI Wen-hao, ZHANG Yu-xiu, WU Hai-ying, ZHANG Jun-na, YE Yu-ling, ZENG Yan, FENG Meng-lian, TIAN Jian-ping. Gene cloning and bioinformatics analysis of chalcone synthase gene from Daphniphyllum calycinum[J]. HUBEI AGRICULTURAL SCIENCES, 2026, 65(5): 205-213.

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