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Cloning and functional analysis of GhDFR1, a key gene of flavonoid synthesis pathway in naturally colored cotton

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Abstract

Background

The naturally colored brown cotton fiber is the most widely used environmentally friendly textile material, which primarily contains proanthocyanidins and their derivatives. Many structural genes in the flavonoid synthesis pathway are known to improve the genetic resources of naturally colored cotton. Among them, DFR is a crucial late enzyme to synthesis both anthocyanins and proanthocyanidins in the plant flavonoid pathway.

Methods

The protein sequences of GhDFRs were analyzed using bioinformatic tools. The expression levels of GhDFRs in various tissues and organs of upland cotton Zongxu1 (ZX1), were analyzed by quantitative real-time PCR, and the expression pattern of GhDFR1 during fiber development of white cotton and brown cotton was analyzed further. The function of GhDFR1 in NCC ZX1 was preliminarily analyzed by virus induced gene silencing (VIGS) technology.

Results

Bioinformatic analysis revealed that GhDFRs sequences in upland cotton genome were extremely conserved. Furthermore, evolutionary tree analysis revealed that the functions of GhDFR1 and GhDFR2, and GhDFR3 and GhDFR4, presented different and shared some similarities. Our study showed GhDFR1 and GhDFR2 were specifically expressed in fibers, while GhDFR3 and GhDFR4 were specifically expressed in petals. GhDFR1 was exclusively expressed in brown cotton fiber at various stages of development and progressively increased with the growth of fiber, but the trend of expression in white cotton was quite the opposite. We silenced GhDFR1 expression in brown cotton fiber using VIGS technology, and observed the VIGS-interference plants. After reducing the expression level of GhDFR1, the period for significant GhDFR1 expression in the developing fibers changed, reducing the content of anthocyanins, and lightening the color of mature cotton fibers.

Conclusion

GhDFR1 was preferentially expressed in brown cotton during fiber development. The timing of GhDFR1 expression for flavonoid synthesis altered, resulting in anthocyanin contents reduced and the fiber color of the GhDFR1i lines lightened. These findings showed the role of GhDFR1 in fiber coloration of NCC and provided a new candidate for NCC genetic improvement.

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Funding

This work was supported by the Natural Science Foundation of Zhejiang Province (Grant No. LZ21C130004) and the National Natural Science Foundation of China (Grant No. U1903204).

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Authors and Affiliations

  1. Plant Genomics and Molecular Improvement of Colored Fiber Laboratory, College of Life Sciences and Medicine, Zhejiang Sci-Tech University, Hangzhou, 310018, People’s Republic of China

    Hongli Zheng, Junye Jiao, Qingqing Niu, Ning Zhu, Yinshuai Huang, Liping Ke & Yuqiang Sun

  2. College of Textile Science and Engineering, Zhejiang Sci-Tech University, Hangzhou, 310018, People’s Republic of China

    Hongli Zheng

  3. China Colored-Cotton (Group) Co., Ltd., Ürümqi, 830011, Xinjiang, People’s Republic of China

    Shouwu Tang & Haifeng Liu

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  1. Hongli Zheng
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  2. Junye Jiao
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  3. Qingqing Niu
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  4. Ning Zhu
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  5. Yinshuai Huang
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Contributions

HZ conducted experiments and wrote the manuscript; JJ and QN took photos and sampling; NZ and YS analyzed the data; ST and HL provided methodology and project administration. LK and YS provided funding, wrote and revised the manuscript. All authors read and agreed to the submitted version of the manuscript.

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Correspondence to Haifeng Liu or Yuqiang Sun.

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Zheng, H., Jiao, J., Niu, Q. et al. Cloning and functional analysis of GhDFR1, a key gene of flavonoid synthesis pathway in naturally colored cotton. Mol Biol Rep 50, 4865–4873 (2023). https://doi.org/10.1007/s11033-023-08420-6

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