Identification and Expression Regulation Analysis of the CsWCOR413 Gene Family in Camellia sinensis

LIU Enbei; WU Yedie; XU Miaomiao; LING Mingxing; PENG Jing; WANG Jie; WANG Xinchao; WANG Lu

doi:10.13305/j.cnki.jts.2026.01.004

Journal of Tea Science >

2026 , Vol. 46 >Issue 1: 1 - 19

DOI: https://doi.org/10.13305/j.cnki.jts.2026.01.004

Research Paper

Identification and Expression Regulation Analysis of the CsWCOR413 Gene Family in Camellia sinensis

LIU Enbei ,
WU Yedie ,
XU Miaomiao ,
LING Mingxing ,
PENG Jing ,
WANG Jie ,
WANG Xinchao ,
WANG Lu

Expand

1. Key Laboratory of Biology, Genetic and Breeding of Special Economic Animals and Plants, Ministry of Agriculture and Rural Affairs, National Center for Tea Plant Improvement, Tea Research Institute, Chinese Academy of Agricultural Sciences, Hangzhou 310008, China;
2. State Key Laboratory of Tea Plant Germplasm Innovation and Resource Utilization, Hangzhou 310008, China

Received date: 2025-09-01

Revised date: 2025-09-22

Online published: 2026-02-06

Fold

Abstract

Low temperature stress significantly impairs the growth and development of tea plants (Camellia sinensis), as well as the yield and quality of tea. The WCOR413 gene family plays an important role in plant cold resistance. However, it has not yet been systematically identified in tea plants. This study employed pan-genome and cold acclimation transcriptome data of tea plants, combined with bioinformatics and RT-qPCR analysis, to identify members of the CsWCOR413 gene family. The results show that the CsWCOR413 family consists of six members (CsWCOR413-1 to CsWCOR413-6), which can be divided into two subclasses based on the conserved domains of the proteins: plasma membrane-localized and internal/thylakoid membrane-localized. Promoter cis-element analysis finds that the CsWCOR413 genes may be regulated by various transcription factors, enabling them to respond to cold, dehydration and ABA signals. RT-qPCR analysis reveals the expression patterns of the CsWCOR413 genes. CsWCOR413-1 was predominantly expressed in leaves, responded to JA and ABA signaling, and was significantly induced by low temperature stress. CsWCOR413-5 responded to short-term cold stress and calcium ion signaling. The transcriptome and Weighted gene co-expression network analysis (WGCNA) indicate that CsWCOR413-1 was significantly upregulated during natural cold acclimation and was strongly co-expressed with CsCBF3, a core transcription factor in cold signaling. Subcellular localization reveals that CsWCOR413-1 was localized in the cell membrane and cytoplasm. Dual-luciferase reporter assay confirms that CsCBF1 and CsCBF3 directly bound to the CsWCOR413-1 promoter and activated its expression. This study systematically elucidated the characteristics and expression profiles of the CsWCOR413 gene family and revealed that CsWCOR413-1 is transcriptionally regulated by CsCBF1 and CsCBF3, suggesting the crucial role of CsWCOR413-1 in tea plant under low temperature stress.

Key words： WCOR413 gene family; expression regulation; low temperature stress; pan-genome; tea plants

Cite this article

LIU Enbei , WU Yedie , XU Miaomiao , LING Mingxing , PENG Jing , WANG Jie , WANG Xinchao , WANG Lu . Identification and Expression Regulation Analysis of the CsWCOR413 Gene Family in Camellia sinensis[J]. Journal of Tea Science, 2026 , 46(1) : 1 -19 . DOI: 10.13305/j.cnki.jts.2026.01.004

References

[1] Ding Y L, Shi Y T, Yang S H.Advances and challenges in uncovering cold tolerance regulatory mechanisms in plants[J]. New Phytologist, 2019, 222(4): 1690-1704.
[2] 陈思琪, 孙敬爽, 麻文俊, 等. 植物低温胁迫调控机制研究进展[J]. 中国农学通报, 2022, 38(17): 51-61.
Chen S Q, Sun J S, Ma W J, et al.Regulation mechanism of low temperature stress on plants: research progress[J]. Chinese Agricultural Science Bulletin, 2022, 38(17): 51-61.
[3] Thomashow M F.So what’s new in the field of plant cold acclimation? Lots![J]. Plant Physiology, 2001, 125(1): 89-93.
[4] Liu B, Wang X Y, Cao Y, et al.Factors affecting freezing tolerance: a comparative transcriptomics study between field and artificial cold acclimations in overwintering evergreens[J]. The Plant Journal, 2020, 103(6): 2279-2300.
[5] Shi Y T, Yang S H.COLD1: a cold sensor in rice[J]. Science China Life Sciences, 2015, 58(4): 409-410.
[6] Hao X Y, Tang H, Wang B, et al.Integrative transcriptional and metabolic analyses provide insights into cold spell response mechanisms in young shoots of the tea plant[J]. Tree Physiology, 2018, 38(11): 1655-1671.
[7] Wang L, Feng X, Yao L N, et al.Characterization of CBL-CIPK signaling complexes and their involvement in cold response in tea plant[J]. Plant Physiology and Biochemistry, 2020, 154: 195-203. doi: 10.1016/j.plaphy.
2020.06.005.
[8] Wang L, Yao L N, Hao X Y, et al.Transcriptional and physiological analyses reveal the association of ROS metabolism with cold tolerance in tea plant[J]. Environmental and Experimental Botany, 2019, 160: 45-58. doi: 10.1016/j.envexpbot.2018.11.011.
[9] Di T M, Wu Y D, Feng X, et al.CIPK11 phosphorylates GSTU23 to promote cold tolerance in Camellia sinensis[J]. Plant, Cell & Environment, 2024, 47(12): 4786-4799.
[10] Di T M, Wu Y D, Wang J, et al.CsCIPK20 improves tea plant cold tolerance by modulating ascorbic acid synthesis through attenuation of CsCSN5-CsVTC1 interaction[J]. Plant, Cell & Environment, 2025, 48(5): 3337-3351.
[11] Wang Y, Jiang C J, Li Y Y, et al.CsICE1 and CsCBF1: two transcription factors involved in cold responses in Camellia sinensis[J]. Plant Cell Reports, 2012, 31(1): 27-34.
[12] Peng J, Li N N, Di T M, et al.The interaction of CsWRKY4 and CsOCP3 with CsICE1 regulates CsCBF1/3 and mediates stress response in tea plant (Camellia sinensis)[J]. Environmental and Experimental Botany, 2022, 199: 104892. doi: 10.1016/j.envexpbot.2022.104892.
[13] Guo X Y, Zhang L, Dong G Q, et al.A novel cold-regulated protein isolated from Saussurea involucrata confers cold and drought tolerance in transgenic tobacco (Nicotiana tabacum)[J]. Plant Science, 2019, 289: 110246. doi: 10.1016/j.plantsci.2019.110246.
[14] Breton G, Danyluk J, Charron J B F, et al. Expression profiling and bioinformatic analyses of a novel stress-regulated multispanning transmembrane protein family from cereals and Arabidopsis[J]. Plant Physiology, 2003, 132(1): 64-74.
[15] Okawa K, Nakayama K, Kakizaki T, et al.Identification and characterization of Cor413im proteins as novel components of the chloroplast inner envelope[J]. Plant, Cell & Environment, 2008, 31(10): 1470-1483.
[16] 丁小玲, 张宁波, 焦淑珍, 等. 山葡萄COR413家族基因克隆及其参与低温胁迫的表达分析[J]. 农业生物技术学报, 2017, 25(3): 366-377.
Ding X L, Zhang N B, Jiao S Z, et al.Cloning and expression analysis of COR413 family genes from Vitis amurensis in response to cold stress[J]. Journal of Agricultural Biotechnology, 2017, 25(3): 366-377.
[17] 王鹏洋, 韩子昂, 翟玉欢, 等. 水曲柳(Fraxinus mandshurica)COR413基因的克隆及表达模式分析[J]. 分子植物育种, 2019, 17(24): 8065-8071.
Wang P Y, Han Z A, Zhai Y H, et al.Cloning and expression pattern analysis of COR413 gene in Fraxinus mandshurica[J]. Molecular Plant Breeding, 2019, 17(24): 8065-8071.
[18] 王鹏飞, 任相亮, 陈曦, 等. 水稻新型潜在冷调节蛋白的生物信息学预测与分析[J]. 安徽农业大学学报, 2011, 38(1): 6-13.
Wang P F, Ren X L, Chen X, et al.Prediction and analysis for new potential rice cold regulated proteins based on bioinformatic method[J]. Journal of Anhui Agricultural University, 2011, 38(1): 6-13.
[19] 苏晨. 拟南芥、水稻膜定位蛋白COR413-PM1通过ABA信号途径调节植物抗逆反应的功能分析[D]. 杨凌: 西北农林科技大学, 2018.
Su C.Functional analysis of membrane localization protein COR413-PM1 regulating plant stress resistance through ABA signaling pathway in Arabidopsis thaliana and rice [D]. Yangling: Northwest A & F University, 2018.
[20] 刘媛君. 黄瓜冷适应蛋白基因CsWCOR413PM的克隆及干扰载体的构建[D]. 天津: 天津农学院, 2020.
Liu Y J.Cloning of cucumber cold adaptation protein gene CsWCOR413PM and construction of its interference vector [D]. Tianjin: Tianjin Agricultural University, 2020.
[21] 王璐瑶, 丁美云, 孙玉婷, 等. 油菜素内酯对低温胁迫下冬小麦wcor413-like基因表达的影响[J]. 麦类作物学报, 2023, 43(4): 463-469.
Wang L Y, Ding M Y, Sun Y T, et al.Regulation of wcor413-like gene expression in winter wheat by low temperature and brassinosteroids[J]. Journal of Triticeae Crops, 2023, 43(4): 463-469.
[22] 秦华, 眭顺照, 李名扬, 等. 蜡梅Chimonanthus praecox(L.) Link COR413蛋白基因(Cpcor413pm1)的分子特性与表达分析[J]. 中国生物化学与分子生物学报, 2006(7): 547-552.
Qin H, Sui S Z, Li M Y, et al.Molecular characterization and expression analysis of a novel COR413 gene from wintersweet [Chimonanthus praecox(L.) Link][J]. Chinese Journal of Biochemistry and Molecular Biology, 2006(7): 547-552.
[23] Ruibal C, Castro A, Fleitas A L, et al.A chloroplast COR413 protein from Physcomitrella patens is required for growth regulation under high light and ABA responses[J]. Frontiers in Plant Science, 2020, 11: 845. doi: 10.3389/fpls.2020.00845.
[24] Danyluk J, Carpentier E, Sarhan F.Identification and characterization of a low temperature regulated gene encoding an actin-binding protein from wheat[J]. FEBS Letters, 1996, 389(3): 324-327.
[25] Zhang H W, Liu W, Wan L Y, et al.Functional analyses of ethylene response factor JERF3 with the aim of improving tolerance to drought and osmotic stress in transgenic rice[J]. Transgenic Research, 2010, 19(5): 809-818.
[26] Gao S M, Zhang H W, Tian Y, et al.Expression of TERF1 in rice regulates expression of stress-responsive genes and enhances tolerance to drought and high-salinity[J]. Plant Cell Reports, 2008, 27(11): 1787-1795.
[27] Lyu J I, Ramekar R, Kim J M, et al.Unraveling the complexity of faba bean (Vicia faba L.) transcriptome to reveal cold-stress-responsive genes using long-read isoform sequencing technology[J]. Scientific Reports, 2021, 11(1): 21094. doi: 10.1038/s41598-021-00506-0.
[28] Chen S, Wang P J, Kong W L, et al.Gene mining and genomics-assisted breeding empowered by the pangenome of tea plant Camellia sinensis[J]. Nature Plants, 2023, 9(12): 1986-1999.
[29] Katoh K, Standley D M.MAFFT multiple sequence alignment software version 7: improvements in performance and usability[J]. Molecular Biology and Evolution, 2013, 30(4): 772-780.
[30] Kumar S, Stecher G, Tamura K.MEGA7: molecular evolutionary genetics analysis version 7.0 for bigger datasets[J]. Molecular Biology and Evolution, 2016, 33(7): 1870-1874.
[31] Price M N, Dehal P S, Arkin A P.FastTree 2: approximately maximum-likelihood trees for large alignments[J]. PLoS One, 2010, 5(3): e9490. doi: 10.1371/journal.pone.0009490.
[32] Hao X Y, Horvath D P, Chao W S, et al.Identification and evaluation of reliable reference genes for quantitative real-time PCR analysis in tea plant (Camellia sinensis (L.) O. Kuntze)[J]. International Journal of Molecular Sciences, 2014, 15(12): 22155-22172.
[33] Livak K J, Schmittgen T D.Analysis of relative gene expression data using real-time quantitative PCR and the <inline-graphic xlink:href="1000-369X-46-1-1/img_1.wmf"/> method[J]. Methods, 2001, 25(4): 402-408.
[34] Wang L, Di T M, Peng J, et al.Comparative metabolomic analysis reveals the involvement of catechins in adaptation mechanism to cold stress in tea plant (Camellia sinensis var. sinensis)[J]. Environmental and Experimental Botany, 2022, 201: 104978. doi: 10.1016/j.envexpbot.2022.104978.
[35] Yao L N, Ding C Q, Hao X Y, et al.CsSWEET1a and CsSWEET17 mediate growth and freezing tolerance by promoting sugar transport across the plasma membrane[J]. Plant & Cell Physiology, 2020, 61(9): 1669-1682.
[36] Li F F, Zhao N, Li Z H, et al.A calmodulin-like protein suppresses RNA silencing and promotes geminivirus infection by degrading SGS3 via the autophagy pathway in Nicotiana benthamiana[J]. PLoS Pathogens, 2017, 13(2): e1006213. doi: 10.1371/journal.ppat.1006213.
[37] Wang J G, Dai S Y, Sun H W, et al.The N-terminal and third transmembrane domain of PsCor413im1 are essential for targeting to chloroplast envelope membrane[J]. Biochemical and Biophysical Research Communications, 2020, 527(4): 929-934.
[38] Liu J L, Wang F T, Yu G, et al.Functional analysis of the maize C-repeat/DRE motif-binding transcription factor CBF3 promoter in response to abiotic stress[J]. International Journal of Molecular Sciences, 2015, 16(6): 12131-12146.
[39] Hu Z, Ban Q Y, Hao J, et al.Genome-wide characterization of the C-repeat binding factor (CBF) gene family involved in the response to abiotic stresses in tea plant (Camellia sinensis)[J]. Frontiers in Plant Science, 2020, 11: 921. doi: 10.3389/fpls.2020.00921.
[40] Panchy N, Lehti-shiu M, Shiu S H. Evolution of gene duplication in plants[J]. Plant Physiology, 2016, 171(4): 2294-2316.
[41] Guo D L, Li Y, Lu H Y, et al.A pangenome reference of wild and cultivated rice[J]. Nature, 2025, 642(8068): 662-671.
[42] Han Z L, Zhang C, Zhang H, et al.CsMYB transcription factors participate in jasmonic acid signal transduction in response to cold stress in tea plant (Camellia sinensis)[J]. Plants, 2022, 11(21): 2869. doi: 10.3390/plants11212869.
[43] Wang Y L, Tong W, Li F D, et al.LUX ARRHYTHMO links CBF pathway and jasmonic acid metabolism to regulate cold tolerance of tea plants[J]. Plant Physiology, 2024, 196(2): 961-978.
[44] Jin J Y, Zhao M Y, Jing T T, et al.(Z)-3-hexenol integrates drought and cold stress signaling by activating abscisic acid glucosylation in tea plants[J]. Plant Physiology, 2023, 193(2): 1491-1507.
[45] 黄玉婷, 钱文俊, 王博, 等. 外源Ca²+及钙离子信号抑制剂对茶树抗寒性的影响[J]. 茶叶科学, 2015, 35(6): 520-526.
Huang Y T, Qian W J, Wang B, et al.Effects of exogenous calcium and inhibitors of calcium signaling transduction pathway on cold resistance of tea plant[J]. Journal of Tea Science, 2015, 35(6): 520-526.
[46] Kopeć P, Rapacz M, Arora R.Post-translational activation of CBF for inducing freezing tolerance[J]. Trends in Plant Science, 2022, 27(5): 415-417.
[47] Zhang X C, Cao X J, Xia Y H, et al.CsCBF5 depletion impairs cold tolerance in tea plants[J]. Plant Science, 2022, 325: 111463. doi: 10.1016/j.plantsci.2022.111463.

Options

Outlines

模态框（Modal）标题

Abstract

Cite this article

References