Acta Horticulturae Sinica ›› 2021, Vol. 48 ›› Issue (10): 1847-1858.doi: 10.16420/j.issn.0513-353x.2021-0090

• Research Papers • Previous Articles     Next Articles

Cloning and Functional Characterization of Chalcone Synthase Genes(CmCHS)from Clivia miniata

LIU Yue1, LI Yueqing2, MENG Xiangyu3, HUANG Jing1, TANG Hao1, LI Yuanheng1, GAO Xiang2, ZHAO Chunli1,*()   

  1. 1College of Horticulture,Jilin Agricultural University,Changchun 130118,China
    2Key Laboratory of Molecular Epigenetics of Ministry of Education,Northeast Normal University,Changchun 130024,China
    3College of Life Science and Bioengineering,Shenyang University,Shenyang 110044,China
  • Received:2021-06-09 Revised:2021-08-30 Online:2021-10-25 Published:2021-11-01
  • Contact: ZHAO Chunli


In order to investigate the structure and function of Chalcone Synthase from Clivia miniata,members of CHS gene family were screened and identified based on the previously constructed transcriptome database. The results showed that four CmCHSsCmCHS1,CmCHS2,CmCHS3 and CmCHS4)genes were cloned in C. miniata. The open reading frames of the four CmCHSs were 1 173,1 170,1 173 and 1 173 bp and encoding 390,389,390,390 respectively. Amino acid sequence that CmCHSs had typical the conserved domains. Phylogenetic tree analysis showed that the four CmCHS genes clustered with the bona-fide CHSs from other plants and belonged to type Ⅲ polyketide synthase(PKS)super family. The results of subcellular localization showed that they were localized in the cytoplasm and nucleus. Gene expressionanalysis showed CmCHS1 and CmCHS2 were highly expressed at the late stage of flower development process,and the transcripts of CmCHS1,CmCHS2 and CmCHS3 were confirmed to the extensively detected in pigmented leaves and red fruit peels. In contrast,the expression level of CmCHS4 gene was relatively low. In order to firmly verified the roles of CmCHSs in the biosynthesis of anthocyanins,prokaryotic expression vectors,pET-32-CmCHS,were constructed and recombinant proteins were prepared and purified,which were subjected to in vitro enzyme activity analysis. Results showed that the four CmCHS recombinant proteins could catalyze coumarinyl-CoA and malonyl-Coenzyme A into naringenin chalcone. This study explored the bioinformatics analysis,gene expression analysis,and functional analysis of CmCHS genes,and laid a theoretical foundation for the genetic transformation and flower color improvement of C. miniata.

Key words: Clivia miniata, flavonoid, anthocyanin, chalcone synthase, enzyme activity

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