Acta Horticulturae Sinica ›› 2021, Vol. 48 ›› Issue (10): 1945-1955.doi: 10.16420/j.issn.0513-353x.2021-0581

• Research Papers • Previous Articles     Next Articles

Cloning and Functional Analysis of CpTT8 Related to Anthocyanin Synthesis in Wintersweet(Chimonanthus praecox

ZHANG Qian1, YANG Nan1,2, SANG Haiyu1, ZHAO Rong1, SONG Xiaoxi1, CHEN Longqing2, XIANG Lin1, ZHAO Kaige1,*()   

  1. 1Key Laboratory of Horticltural Plant Biology,Ministy of Edtcation,College of Horticulture and Forestry Sciences,Huazhong Agricultural University,Wuhan 430070,China
    2College of Landscape Architecture and Horticulture Sciences,Southwest Forestry University,Kunming 650224,China
  • Received:2021-06-18 Revised:2021-09-09 Online:2021-10-25 Published:2021-11-01
  • Contact: ZHAO Kaige


The full-length coding sequence of a bHLH transcription factor,named CpTT8,was cloned from Chimonanthus praecox‘H29’. Studies related to this transcription factor were carried out,including bioinformatics characteristics,spatiotemporal expression pattern analysis,gene function test,protein interactions,and protein-DNA interaction. Sequence analysis showed that the ORF of CpTT8 consists of 2 130 bp encoding 709 amino acids,and CpTT8 contains a bHLH domain. CpTT8 shows the highest sequence similarity to PpbHLH3 from peach and MdbHLH3 from apple. The results of quantitative RT-PCR revealed that the expression level of CpTT8 was highest in flower bud at the early stage of floral development. It was also expressed in roots,stems,leaves,and fruits,with the highest level in fruits. Overexpression of CpTT8 in Arabidopsis Col-0 and tt8 facilitated the accumulation of anthocyanins and proanthocyanidins,indicating that CpTT8 is a critical positive regulatory transcription factor in the anthocyanin biosynthesis pathway. Yeast two-hybrid assay and dual-luciferase analysis illustrated that CpTT8 needs to interact with MYB transcription factors to activate its downstream structural gene ANS.

Key words: Chimonanthus praecox, anthocyanin, CpTT8, functional identification, interaction

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