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Molecular and biochemical characterization of two 4-coumarate: CoA ligase genes in tea plant (Camellia sinensis)

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Two 4-coumarate: CoA ligase genes in tea plant involved in phenylpropanoids biosynthesis and response to environmental stresses.

Abstract

Tea plant is rich in flavonoids benefiting human health. Lignin is essential for tea plant growth. Both flavonoids and lignin defend plants from stresses. The biosynthesis of lignin and flavonoids shares a key intermediate, 4-coumaroyl-CoA, which is formed from 4-coumaric acid catalyzed by 4-coumaric acid: CoA ligase (4CL). Herein, we report two 4CL paralogs from tea plant, Cs4CL1 and Cs4CL2, which are a member of class I and II of this gene family, respectively. Cs4CL1 was mainly expressed in roots and stems, while Cs4CL2 was mainly expressed in leaves. The promoter of Cs4CL1 had AC, nine types of light sensitive (LSE), four types of stress-inducible (SIE), and two types of meristem-specific elements (MSE). The promoter of Cs4CL2 also had AC and nine types of LSEs, but only had two types of SIEs and did not have MSEs. In addition, the LSEs varied in the two promoters. Based on the different features of regulatory elements, three stress treatments were tested to understand their expression responses to different conditions. The resulting data indicated that the expression of Cs4CL1 was sensitive to mechanical wounding, while the expression of Cs4CL2 was UV-B-inducible. Enzymatic assays showed that both recombinant Cs4CL1 and Cs4CL2 transformed 4-coumaric acid (CM), ferulic acid (FR), and caffeic acid (CF) to their corresponding CoA ethers. Kinetic analysis indicated that the recombinant Cs4CL1 preferred to catalyze CF, while the recombinant Cs4CL2 favored to catalyze CM. The overexpression of both Cs4CL1 and Cs4CL2 increased the levels of chlorogenic acid and total lignin in transgenic tobacco seedlings. In addition, the overexpression of Cs4CL2 consistently increased the levels of three flavonoid compounds. These findings indicate the differences of Cs4CL1 and Cs4CL2 in the phenylpropanoid metabolism.

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Funding

Funding was provided by Natural Science Foundation of China (Grant Nos. 31870677 and 31870676).

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Authors and Affiliations

  1. State Key Laboratory of Tea Plant Biochemistry and Utilization, Anhui Agricultural University, Hefei, 230036, China

    Mingzhuo Li, Yanzhi Li, Xiaolan Jiang, Yajun Liu & Tao Xia

  2. School of Life Science, Anhui Agricultural University, Hefei, 230036, China

    Lili Guo, Yeru Wang & Liping Gao

  3. Department of Plant and Microbial Biology, North Carolina State University, Raleigh, NC, 27695, USA

    Mingzhuo Li & De-Yu Xie

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  1. Mingzhuo Li
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  2. Lili Guo
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  3. Yeru Wang
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  4. Yanzhi Li
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  5. Xiaolan Jiang
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  6. Yajun Liu
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  7. De-Yu Xie
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  8. Liping Gao
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  9. Tao Xia
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Contributions

ML, LG, and TX conceived the experiments. ML, LG, YW, XJ and YL performed the experiments. ML and DX prepared the manuscript. All authors read and approved the manuscript.

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Correspondence to De-Yu Xie, Liping Gao or Tao Xia.

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Li, M., Guo, L., Wang, Y. et al. Molecular and biochemical characterization of two 4-coumarate: CoA ligase genes in tea plant (Camellia sinensis). Plant Mol Biol 109, 579–593 (2022). https://doi.org/10.1007/s11103-022-01269-6

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