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Flavonoid 3′-hydroxylase of Camellia nitidissima Chi. promotes the synthesis of polyphenols better than flavonoids

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Abstract

Camellia nitidissima Chi. is an ornamental plant of the genus Camellia L. Its flowers contain a lot of flavonoids and polyphenols. Flavonoid 3′-hydroxylase (F3′H) plays an important role in the synthesis of flavonoids, polyphenols and anthocyanins. We used PCR amplification, quantitative PCR, High-performance liquid chromatography, subcellular localization, and agrobacterium-mediated leaf disk method to study the the function of CnF3′H. The full length of CnF3′H was 1859 bp (GenBank code: HQ290518.1), with an open reading frame of 1577 bp, and encoded 518 amino acid. A phylogenetic tree analysis showed that CnF3′H was closely related to Camellia sinensis L. and C. sinensis cultivar Zhonghuang. CnF3′H was expressed in flowers, leaves, fruits, sepals, petals and stamens of C. nitidissima, and during the flowering process the expression level in flower decreased initially and then increased. CnF3′H expression was significantly positive correlated with polyphenol contents in C. nitidissima. A CnF3′H-EGFP expression vector was constructed to do the subcellular localization, we found that CnF3′H was obviously localized in the nuclear envelope and cytomembrane. In transgenic tobacco flowers, the total polyphenol content and various polyphenol constituents were significantly increased with high CnF3′H expression level, while total flavonoid contents and some flavonol constituents were increased slightly. These findings suggest that CnF3′H promotes the synthesis of polyphenols better than flavonoids.

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Data Availability

All data are presented in the manuscript. Camellia nitidissima Chi. tissues were collected from the National Camellia Germplasm Resource Bank (Guangxi, China).

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Acknowledgements

The authors thank the members of the National Forestry and Grassland Administration Economic Forest Product Quality Supervision, Inspection and Testing Center (Hangzhou) for their assistance in condition establishment and detection of high-performance liquid phase.

Funding

This work was supported by the National Key R&D Program of China (2019YFD1001005), the special funds for basic scientific research expenses of public welfare research institutes of the Chinese academy of forestry (CAFYBB2017ZF001), The National key projects for international scientific and technological innovation cooperation among governments (2016YFE0126100), The National Natural Science Foundation of China (No. 31860228), the Key projects of Natural Science Foundation of the Guangxi Zhuang Autonomous Region (2018GXNSFDA281007), and Scientific Research Foundation of Yulin Normal University for high-level talents (G2018025).

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Authors and Affiliations

  1. Research Institute of Subtropical Forestry, Chinese Academy of Forestry, Hangzhou, 311400, China

    Lina Jiang, Zhengqi Fan, Ran Tong, Hengfu Yin & Jiyuan Li

  2. State Key Laboratory of Tree Genetics and Breeding, Research Institute of Subtropical Forestry, Chinese Academy of Forestry, Hangzhou, 311400, China

    Lina Jiang & Hengfu Yin

  3. Key Laboratory of Forest Genetics and Breeding, Research Institute of Subtropical Forestry, Chinese Academy of Forestry, Hangzhou, 311400, China

    Lina Jiang, Zhengqi Fan, Hengfu Yin & Jiyuan Li

  4. College of Architecture and Urban Planning, Fujian University of Technology, Fuzhou, 350117, China

    Xingwen Zhou

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  1. Lina Jiang
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  3. Ran Tong
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Contributions

JL finished the main experimental contentc, wrote and modified the paper. FZ and TR participated in the experiment and data collection together. ZX provided the foundation and thought for the preliminary study. LJ was the architect and director of the project. YH was responsible for the experimental design, and guided the writing and modification of the paper. All authors read and agree to the final manuscript.

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Correspondence to Jiyuan Li or Xingwen Zhou.

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Jiang, L., Fan, Z., Tong, R. et al. Flavonoid 3′-hydroxylase of Camellia nitidissima Chi. promotes the synthesis of polyphenols better than flavonoids. Mol Biol Rep 48, 3903–3912 (2021). https://doi.org/10.1007/s11033-021-06345-6

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