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Regulation of Anthocyanin Accumulation by a Transcription Factor LcTT8 From Lonicera caerulea L.

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Abstract

The fruit of Lonicera caerulea L. is rich in anthocyanin, which has beneficial effects on human health. However, there is no research report on the regulation mechanism of anthocyanin synthesis in Lonicera caerulea L. TT8 gene-encoded protein belongs to bHLH transcription factor, and it has been reported to regulate anthocyanin synthesis. In the study, we cloned TT8 gene from Lonicera caerulea L. named “Berel” and transformed into tobacco (Nicotiana tabacum) to characterize its function. The open-reading frame of LcTT8 was 2049 bp, encoding a protein of 682 amino acids. The phylogenetic tree analysis showed that LcTT8 protein had the highest homology with TT8 in Paeonia suffruticosa. The analysis of expression in different tissues showed that the expression level of LcTT8 was higher in fruits than that in flowers, leaves, stems, and roots, and the expression level reached the peak at the early stage of fruit color change. The subcellular localization results showed that the LcTT8 protein was mainly localized in the nucleus. A total of ten LcTT8 overexpression transgenic tobacco were obtained. The ELISA results revealed that the anthocyanin content in the transgenic tobacco leaves was increased by 36.84% compared with the wild-type tobacco. The qPCR results showed that the overexpression of LcTT8 caused the up-expression of key genes of anthocyanin synthesis, including F3H, DFR, and ANS. Similarly, the enzyme activities of DFR and ANS were significantly increased in the transgenic lines (P < 0.05). Our research reveals the expression patterns and molecular characteristic of LcTT8 gene in anthocyanin synthesis, providing new possibilities for the breeding of blue honeysuckle and other fruit plants with high anthocyanin accumulation.

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Funding

This research was supported by the National Key Research and Development Project (2016YFC0500304), National Key R&D Program of China (2018YFD100200), Young Talents” Project of Northeast Agricultural University (19QC05), Heilongjiang Province Postdoctoral Startup Fund Postdoctoral Grant (LBH-Q17029), Natural Science Foundation of Heilongjiang Province of China (C2017020), and Natural Science Foundation of Heilongjiang Province of China (C2017015, C2017015).

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Author notes

  1. Pei Liu and Huixin Gang contributed equally to this work.

Authors and Affiliations

  1. National-Local Joint Engineering Research Center for Development and Utilization of Small Fruits in Cold Regions, Northeast Agricultural University, Harbin, 150030, China

    Pei Liu, Huixin Gang, Huayu Liu, Dong Qin, Yan Zhang & Junwei Huo

  2. College of Horticulture and Landscape, Northeast Agricultural University, Harbin, 150030, China

    Pei Liu, Huixin Gang, Huayu Liu, Dong Qin, Yan Zhang & Junwei Huo

Authors
  1. Pei Liu
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  2. Huixin Gang
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  3. Huayu Liu
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  4. Dong Qin
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  5. Yan Zhang
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  6. Junwei Huo
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Correspondence to Dong Qin or Junwei Huo.

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Key message

The expression patterns and molecular characteristic of LcTT8 gene of bHLH family were studied in anthocyanin synthesis, providing new possibilities for the breeding of blue honeysuckle and other fruit plants with high anthocyanin accumulation.

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Liu, P., Gang, H., Liu, H. et al. Regulation of Anthocyanin Accumulation by a Transcription Factor LcTT8 From Lonicera caerulea L.. Plant Mol Biol Rep 39, 125–136 (2021). https://doi.org/10.1007/s11105-020-01229-8

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