CmTCP9/12/25-CmSAUR1 module assists trehalose-induced cold tolerance in melon seedlings

doi:10.1016/j.jia.2026.05.057

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CmTCP9/12/25-CmSAUR1 module assists trehalose-induced cold tolerance in melon seedlings

Adan Liang^1,2*, Yuqing Han³*, Yujie Zhang^1,2*, Jiahui Tian^1,2*, Dongdong Xu^1,2, Li Shao^1,2, Hongyi Zhang^1,2, Haiming Li^1,2, Xueling Ye^1,2, Tao Liu^1,2#, Hongyan Qi^1,2#

¹College of Horticulture, Shenyang Agricultural University, Shenyang, 110866, China

²Key Laboratory of Protected Horticulture of Education of Ministry and Liaoning Province, Northern National & Local Joint Engineering Research Center of Horticultural Facilities Design and Application Technology, Shenyang, Liaoning, 110866, China

³Horticultural Research Institute and Shanghai Key Lab of Protected Horticultural Technology, Shanghai Academy of Agricultural Sciences, Shanghai 201403, China

Highlights

IAA-CmSAUR1 positively regulates the cold tolerance of melon seedlings

Tre induced CmTCP9/12/25 positively regulate the transcription of CmSAUR1

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摘要

冷害限制了甜瓜正常生长发育。我们前期研究表明外施海藻糖（Tre）能缓解冷胁迫对甜瓜幼苗的损伤，但其具体作用机制尚不明晰。在本研究中，Tre预处理增加了吲哚-3-乙酸（IAA）的含量，并上调了 IAA早期响应因子（small auxin up RNA gene, CmSAUR1）基因表达。当采用N-1-萘基酞氨酸抑制IAA信号运输或采用VIGS技术沉默 CmSAUR1 会降低甜瓜幼苗超氧化物歧化酶、抗坏血酸过氧化物酶和谷胱甘肽还原酶的活性，加重了甜瓜幼苗冷胁迫损伤，并削弱了Tre的缓解作用。通过对转录组数据和CmSAUR1基因启动子的分析，挖掘出CmSAUR1上游转录因子CmTCP9/12/25。通过蛋白质-DNA互作试验进一步证实了CmTCP9/12/25能与CmSAUR1的启动子结合并激发其表达。通过酵母双杂交、荧光素酶互补成像和双分子荧光互补试验进一步证实了CmTCP25能分别与CmTCP9和CmTCP12形成蛋白复合体发挥作用。沉默CmTCP9/12/25基因下调了CmSAUR1的表达，加剧了甜瓜幼苗的冷损伤，并削弱了Tre的缓解效应。综上，Tre预处理诱导了CmTCP9/12/25的上调表达，CmTCP9/12/25能与CmSAUR1启动子结合并激活其表达，最终提高了甜瓜幼苗耐冷性。

Abstract

Cold stress negatively affects melon growth and development. Previous studies have shown that exogenous trehalose (Tre) alleviates the cold damage in melon seedlings; however, the specific mechanism is not fully clarified. Here, Tre treatment increased the indole-3-acetic (IAA) content and upregulated an early response factor of IAA (small auxin up RNA gene, CmSAUR1) gene expression. Inhibition of IAA signal transport or silencing of CmSAUR1 decreased the activities of superoxide dismutase, ascorbate peroxidase and glutathione reductase. This led to increased susceptibility to cold damage in melon seedlings and diminished the beneficial impact of Tre. Through the analysis of transcriptome and promoter of CmSAUR1, the upstream transcription factor CmTCP9/12/25 of CmSAUR1 was excavated. Protein–DNA interaction experiments further verified that CmTCP9/12/25 could transcriptionally regulate CmSAUR1 expression. Yeast two-hybrid, luciferase complementation imaging and bimolecular fluorescence complementation experiments further confirmed that CmTCP9/12 interacted with CmTCP25 to form protein complexes. Silencing CmTCP9/12/25 genes significantly aggravated the cold damage of melon seedlings and weakened the efficacy of Tre. All of them downregulated the expression of CmSAUR1. In conclusion, application of Tre induced the upregulation of CmTCP9/12/25, and CmTCP9/12/25 could bind to the CmSAUR1 promoter and activate its expression. These eventually increased the cold tolerance of melon seedlings.

Keywords: Cucumis melo teosinte branched1/cycloidea/proliferating cell factor small auxin up RNA gene 1 cold stress, trehalose

Online: 30 May 2026

Fund:

This work was supported by grants from the Project funded by the Project funded by the Liao Ning Revitalization Talents Program (XLYC2402014), the National Natural Science Foundation of Liaoning (2024-MSLH-406), the China Agriculture Research System of MOF and MARA (CARS-25), the National Natural Science Foundation of China (U20A2044) and the Basic Research Project of Liaoning Provincial Department of Education (JYTZD2023117).

About author: 1Adan Liang, E-mail: 2656478680@qq.com; * These authors contributed equally to this study. #Correspondence Tao Liu, Tel: +86-24-88487143, E-mail: taoliu@syau.edu.cn; Hongyan Qi, Tel: +86-24-88487144, E-mail: qihongyan@syau.edu.cn.

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Adan Liang, Yuqing Han, Yujie Zhang, Jiahui Tian, Dongdong Xu, Li Shao, Hongyi Zhang, Haiming Li, Xueling Ye, Tao Liu, Hongyan Qi. 2026. CmTCP9/12/25-CmSAUR1 module assists trehalose-induced cold tolerance in melon seedlings. Journal of Integrative Agriculture, Doi:10.1016/j.jia.2026.05.057

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