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Characterization of Rosa chinensis CBF Genes and the Function of RcCBF6 in Cold Tolerance

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Abstract

As a class of AP2 family transcription factors, C-repeat binding factor (CBF) gene family plays an important role in resisting low-temperature stress and improving cold tolerance of plants. Rosa chinensis is an important horticultural and ornamental plant, but little is known about the molecular characteristics of CBF genes in R. chinensis. In our study, six RcCBFs members were identified at R. chinensis whole genome, including one member RcCBF1 on Chromosome 1 and other five members RcCBF2-6 on Chromosome 7. The analysis of cis-acting elements indicated that there were various cis-acting elements related to stress, development, hormone, and light. The expression profiles showed that most of RcCBFs were mainly expressed in root, and the expression levels of RcCBFs were significantly induced by low-temperature stress, especially RcCBF6. To verify the function of RcCBF6, we generated its overexpressing transgenic lines in Arabidopsis thaliana. The RcCBF6-overexpressing plants exhibited higher tolerance to cold stress as evidenced by a better growth and higher antioxidative enzyme activities than the wild-type plants. Furthermore, the expression levels of some cold-response genes were up-regulated in the transgenic plants, such as KIN1, RD29A, LTP3, and GOLS3. Our study contributes to a better understanding of RcCBF gene family and provides a foundation for the further functional research of RcCBFs.

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Acknowledgements

This work was supported by the Fundamental Research Funds of Chinese Academy of Forestry (CAFYBB2016MA008, CAFYBB2017ZD005 and CAFYBB2019ZY003).

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Authors and Affiliations

  1. Experimental Center of Forestry in North China, Chinese Academy of Forestry, Beijing, 102300, China

    Jianbo Li, Guangshun Zheng, Shaofeng Li, Zhimin Tang, Jingshuang Sun & Xuebing Xin

  2. State Key Laboratory of Tree Genetics and Breeding, Chinese Academy of Forestry, Beijing, 100091, China

    Jianbo Li

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  1. Jianbo Li
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  2. Guangshun Zheng
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  3. Shaofeng Li
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  4. Zhimin Tang
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Contributions

JL, JS, and XX designed the research. JL, GZ, SL, and JS performed the experiments and data analysis. JL wrote the manuscript. ZT, JS, and XX contributed with valuable discussions. All authors read and approved the final manuscript.

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Correspondence to Jianbo Li, Jingshuang Sun or Xuebing Xin.

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Electronic supplementary material

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Supplementary file1 Fig. S1. Prediction of 3D structure of RcCBF proteins. (TIF 10862 kb)

12374_2020_9248_MOESM2_ESM.tif

Supplementary file2 Fig. S2. Distribution of the conserved motifs in RcCBF proteins from R. chinensis, A. thaliana, O. sativa, P. trichocarpa, and C. sinensis. Ten motifs were marked with different colors. (TIF 1753 kb)

12374_2020_9248_MOESM3_ESM.tif

Supplementary file3 Fig. S3. Expression levels of 35 cold-responsive genes in the WT and RcCBF6-overexpressing plants. (TIF 5051 kb)

12374_2020_9248_MOESM4_ESM.tif

Supplementary file4 Fig. S4. Various cis-acting elements in the promoter (2.0 kb upstream of the translation initiation sites) of AtCBF genes. The statistics of total number of RcCBFs contain various cis-acting elements. (TIF 3954 kb)

Supplementary file5 (XLSX 9 kb)

Supplementary file6 (XLSX 11 kb)

Supplementary file7 (XLSX 13 kb)

Supplementary file8 (XLSX 13 kb)

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Li, J., Zheng, G., Li, S. et al. Characterization of Rosa chinensis CBF Genes and the Function of RcCBF6 in Cold Tolerance. J. Plant Biol. 63, 267–278 (2020). https://doi.org/10.1007/s12374-020-09248-4

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