Cloning of the StTrihelix30 Gene in Potatoes and Functional Analysis Under Low-temperature Stress

doi:10.16420/j.issn.0513-353x.2025-0363

Acta Horticulturae Sinica ›› 2026, Vol. 53 ›› Issue (8): 2306-2318.doi: 10.16420/j.issn.0513-353x.2025-0363

• Genetic & Breeding · Germplasm Resources · Molecular Biology • Previous Articles Next Articles

Cloning of the StTrihelix30 Gene in Potatoes and Functional Analysis Under Low-temperature Stress

YANG Hongyu, WANG Yan, LIU Taotao, YAO Wenxia, FAN Xiangjun, YU Bin, SHI Guiying^*()

College of Horticulture， Gansu Agricultural University， Lanzhou 730070， China

Received:2026-04-07 Revised:2026-06-30 Online:2026-08-25 Published:2026-08-25
Contact: SHI Guiying

Abstract

Abstract:

In order to explore the functional divergence of the Trihelix gene from potato（Solanum tuberosum L.）in low temperature response，the previously identified potato low-temperature-responsive gene StTrihelix30 was overexpressed in Arabidopsis thaliana，and the low-temperature tolerance physiological indices of the overexpression lines were determined. The results showed that StTrihelix30 is non-toxic and exhibits self-activation activity. In cold tolerance assays，transgenic Arabidopsis lines overexpressing StTrihelix30 displayed enhanced cold resistance under low-temperature stress，as evidenced by a 27.3% reduction in relative electrical conductivity and a 34.5% decrease in malondialdehyde（MDA）content compared with the wild type. Furthermore，the activities of antioxidant enzymes were significantly increased，with superoxide dismutase（SOD），peroxidase（POD），and catalase（CAT）activities rising to 1.8-fold，2.1-fold，and 1.5-fold those of the wild type，respectively，indicating a markedly enhanced capacity to scavenge reactive oxygen species（ROS）. In conclusion，StTrihelix30 enhances plant adaptability to low-temperature stress by improving its antioxidant capacity.

Key words: Solanum tuberosum L., Trihelix transcription factor family, low-temperature stress, gene cloning

YANG Hongyu, WANG Yan, LIU Taotao, YAO Wenxia, FAN Xiangjun, YU Bin, SHI Guiying. Cloning of the StTrihelix30 Gene in Potatoes and Functional Analysis Under Low-temperature Stress[J]. Acta Horticulturae Sinica, 2026, 53(8): 2306-2318.

Figures/Tables 12

References 22

[23]	Wise R R, Naylor A W. 1987. Chilling-enhanced photooxidation:evidence for the role of singlet oxygen and superoxide in the breakdown of pigments and endogenous antioxidants. Plant Physiology, 83 (2):278-282. doi: 10.1104/pp.83.2.278 pmid: 16665236
[24]	Xie Z M, Zou H F, Lei G, Wei W, Zhou Q Y, Niu C F, Chen S Y. 2009. Soybean Trihelix transcription factors GmGT-2A and GmGT-2B improve plant tolerance to abiotic stresses in transgenic Arabidopsis. PLoS ONE, 4 (9):e6898.
[25]	Yang H Y, Wang Y, Liu T T, Yao W X, Fan X J, Yu B, Shi G Y. 2025. Genome-wide identification of potato Trihelix gene family and its response to different abiotic stresses. BMC Plant Biology, 25 (1):690. doi: 10.1186/s12870-025-06437-6 pmid: 40410672
[26]	Yang H Q, Wu F H, Cheng J Y. 2011. Reduced chilling injury in cucumber by nitric oxide and the antioxidant response. Food Chemistry, 127 (3):1237-1242. doi: 10.1016/j.foodchem.2011.02.011 pmid: 25214120
[27]	Yang J, Tang Z X, Yang W Y, Huang Q H, Wang Y Q, Huang M F, Zhang J. 2023. Genome-wide characterization and identification of Trihelix transcription factors and expression profiling in response to abiotic stresses in Chinese Willow(Salix matsudana Koidz). Frontiers in Plant Science, 14:1125519. doi: 10.3389/fpls.2023.1125519 URL
[28]	Yu Chuying. 2018. Identification of QTLs for cold resistance in tomato and functional analysis of cold response gene ShCIGT[Ph. D. Dissertation]. Huazhong Agricultural University. (in Chinese)
	余楚英. 2018. 番茄抗寒性QTL鉴定及冷响应基因ShCIGT的功能解析[博士论文]. 华中农业大学.
[29]	Zhang C Z, Yang Z M, Tang D, Zhu Y H, Wang P, Li D W, Huang S W. 2021. Genome design of hybrid potato. Cell, 184 (15):3873-3883. doi: 10.1016/j.cell.2021.06.006 pmid: 34171306
[1]	Chang D C, Sohn H B, Cho J H, Ju S I, Jin Y I, Do G R, Kim S J, Cho H M, Lee Y B. 2014. Freezing and frost damage of potato plants:a case study on growth recovery,yield response,and quality changes. Potato Research, 57 (2):99-110. doi: 10.1007/s11540-014-9253-5 URL
[2]	Cheng X, Xiong R, Yan H, Gao Y M, Liu H L, Wu M, Xiang Y. 2019. The trihelix family of transcription factors:functional and evolutionary analysis in Moso bamboo(Phyllostachys edulis). BMC Plant Biology, 19 (1):154. doi: 10.1186/s12870-019-1744-8
[3]	Dehesh K, Smith L G, Tepperman J M, Quail P H. 1995. Twin autonomous bipartite nuclear localization signals direct nuclear import of GT-2. The Plant Journal, 8 (1):25-36.
[4]	Gao Hongyan. 2019. Whole genome analysis of the bamboo family and functional studies of PheGT8 and PheGT16[M. D. Dissertation]. Beijing: Chinese Academy of Forestry. (in Chinese)
	高红艳. 2019. 毛竹三螺旋家族全基因组分析及PheGT8和PheGT16的功能研究[硕士论文]. 北京: 中国林业科学研究院.
[5]	Greb-Markiewicz B, Kazana W, Zarębski M, Ożyhar A. 2019. The subcellular localization of bHLH transcription factor TCF 4 is mediated by multiple nuclear localization and nuclear export signals. Scientific Reports, 9 (1):15629. doi: 10.1038/s41598-019-52239-w pmid: 31666615
[6]	Gu Y, He L, Zhao C, Zhao H J, Wang F, Yan B W, Gao Y Q, Li Z T, Yang K J, Xu J Y. 2017. Biochemical and transcriptional regulation of membrane lipid metabolism in maize leaves under low temperature. Frontiers in Plant Science, 8:2053. doi: 10.3389/fpls.2017.02053 pmid: 29250095
[7]	Hu Yadan, Wu Guoqiang, Liu Chen, Wei Ming. 2024. The role of MYB transcription factors in regulating plant responses to environmental stress. Biotechnology Bulletin, 40 (6):5-22. (in Chinese) doi: 10.13560/j.cnki.biotech.bull.1985.2023-1186
	胡雅丹, 伍国强, 刘晨, 魏明. 2024. MYB 转录因子在调控植物响应逆境胁迫中的作用. 生物技术通报, 40 (6):5-22. doi: 10.13560/j.cnki.biotech.bull.1985.2023-1186
[8]	Kaplan-Levy R N, Brewer P B, Quon T, Smyth D R. 2012. The trihelix family of transcription factors-light,stress and development. Trends in Plant Science, 17 (3):163-171. doi: 10.1016/j.tplants.2011.12.002 pmid: 22236699
[9]	Khalid M H B, Raza M A, Yu H Q, Khan I, Sun F A, Feng L Y, Qu J T, Fu F L, Li W C. 2019. Expression,subcellular localization,and interactions of CPK family genes in maize. International Journal of Molecular Sciences, 20 (24):doi:10.3390/ijms20246173.
[10]	Li H W, Luo W B, Ji R C, Xu Y Q, Xu G C, Qiu S X, Tang H. 2021a. A comparative proteomic study of cold responses in potato leaves. Heliyon, 7 (2):e06002. doi: 10.1016/j.heliyon.2021.e06002 URL
[11]	Li K, Duan L, Zhang Y, Shi M X, Chen S S, Yang M F, Ding Y Q, Peng Y S, Dong Y B, Yang H, Li Z H, Zhang L Y, Fan Y, Ren M J. 2021b. Genome-wide identification and expression profile analysis of trihelix transcription factor family genes in response to abiotic stress in sorghum[Sorghum bicolor(L.) Moench]. BMC Genomics, 22 (1):738.
[12]	Li Yanmei. 2025. The molecular mechanism of MdbHLH162-like transcription factor regulating iron absorption in apple rootstocks[Ph. D. Dissertation]. Lanzhou: Gansu Agricultural University. (in Chinese)
	李艳梅. 2025. MdbHLH162-like转录因子调控苹果砧木铁吸收的分子机制[博士论文]. 兰州: 甘肃农业大学.
[13]	Lian Xuan, Leng Can, Li Yinruizhi, Wu La, Xu Lixin, Dong Di, Tan Penghui, Chao Yuehui. 2019. Cloning,expression and subcellular localization analysis of MtGT1 gene from Peucedanum lanceolatum. Molecular Plant Breeding, 17 (13):4241-4248. (in Chinese)
	连燚, 冷暖, 李殷睿智, 乌拉, 许立新, 董迪, 檀鹏辉, 晁跃辉. 2019. 蒺藜苜蓿MtGT1基因克隆、表达及亚细胞定位分析. 分子植物育种, 17 (13):4241-4248.
[14]	Liang Shumin, Luo Donglan, Cheng Yujin, Yu Le’an, Yang Yajie, Chen Jianye, Lu Wangjin, Kun Jianfei. 2017. Isolation and characterization of the banana trihelix transcription factor MaGTL1a. Journal of Fruit Science, 34 (8):935-945. (in Chinese)
	梁淑敏, 罗冬兰, 程玉瑾, 俞乐安, 杨亚杰, 陈建业, 陆旺金, 邝健飞. 2017. 香蕉trihelix转录因子MaGTL1a的分离及特性. 果树学报, 34 (8):935-945.
[15]	Liu W, Zhang Y, Li W, Lin Y, Wang C, Xu R, Zhang L. 2020. Genome-wide characterization and expression analysis of soybean Trihelix gene family. PeerJ, 8:e8753. doi: 10.7717/peerj.8753 URL
[16]	Liu Yong, Wang Zeqiong, Gong Linzhong, Wang Furong, Wang Huiyang, Ai Xiaoyan, He Huaping. 2020. Cloning and functional analysis of the apetolevulinic acid response factor PpERF1a. Acta Horticulturae Sinica, 47 (6):1165-1171. (in Chinese)
	刘勇, 王泽琼, 龚林忠, 王富荣, 王会良, 艾小艳, 何华平. 2020. 桃乙烯应答因子PpERF1a的克隆与功能分析. 园艺学报, 47 (6):1165-1171. doi: 10.16420/j.issn.0513-353x.2019-0872
[17]	Pham G M, Hamilton J P, Wood J C, Burke J T, Zhao H N, Vaillancourt B, Ou S J, Jiang J M, Buell C R. 2020. Construction of a chromosome-scale long-read reference genome assembly for potato. GigaScience, 9 (9):1-11.
[18]	Qin Y, Ma X, Yu G, Wang Q I, Wang L, Kong L, Kim W, Wang H W. 2014. Evolutionary history of trihelix family and their functional diversification. DNA Research, 21 (5):499-510. doi: 10.1093/dnares/dsu016 pmid: 24864043
[19]	Ruan Mengbin, Peng Ming. 2011. Research progress on genes related to plant responses to abiotic stress. Tropical Biology Journal, 2 (4):364-372. (in Chinese)
	阮孟斌, 彭明. 2011. 植物响应非生物胁迫相关基因的研究进展. 热带生物学报, 2 (4):364-372.
[30]	Zhou Q, Tang D, Huang W, Yang Z M, Zhang Y, Hamilton J P, Huang S W. 2020. Haplotype-resolved genome analyses of a heterozygous diploid potato. Nature Genetics, 52 (10):1018-1023. doi: 10.1038/s41588-020-0699-x pmid: 32989320
[20]	Shafi A, Chauhan R, Gill T, Swarnkar K M, Sreenivasulu Y, Kumar S, Kumar N, Shankar R, Ahuja P S, Singh A K. 2015. Expression of SOD and APX genes positively regulates secondary cell wall biosynthesis and promotes plant growth and yield in Arabidopsis under salt stress. Plant Molecular Biology, 87 (6):615-631. doi: 10.1007/s11103-015-0301-6 URL
[21]	Shafi A, Pal A K, Sharma V, Kalia S, Kumar S, Ahuja P S, Singh A K. 2017. Transgenic potato plants overexpressing SOD and APX exhibit enhanced lignification and starch biosynthesis with improved salt stress tolerance. Plant Molecular Biology Reporter, 35 (5):504-518. doi: 10.1007/s11105-017-1041-3 URL
[22]	Tang D, Jia Y, Zhang J, Li H B, Cheng L, Wang P, Bao A G, Liu Z H, Feng S S, Zhu X J, Li D W, Zhu G T, Wang H R, Zhou Y, Zhou Y F, Bryan G J, Buell C R, Huang S W. 2022. Genome evolution and diversity of wild and cultivated potatoes. Nature, 606 (7914):535-541. doi: 10.1038/s41586-022-04822-x

用途Purpose	引物名称 Primer name	引物序列（5′-3′）Primers sequence
瞬时表达/过表达载体 Transient expression / overexpression vector	StTrihelix30-F	TTTGGAGAGGACACGCTCGAGATGCTGGAAAGTTCTGTTTTGTTG
瞬时表达/过表达载体 Transient expression / overexpression vector	StTrihelix30-R	GCCCTTGCTCACCATGAATTCACATTCTGTTCTTTCTGATGCTTGTT
BD载体 BD carrier	StTrihelix30-F	AGGCCGAATTCCCGGGGATCCTTATGCTGGAAAGTTCTGTTTTGTTG
BD载体 BD carrier	StTrihelix30-R	CCGCTGCAGGTCGACGGATCCTCAACATTCTGTTCTTTCTGATGCT

用途Purpose	引物名称 Primer name	引物序列（5′-3′）Primers sequence
瞬时表达/过表达载体 Transient expression / overexpression vector	StTrihelix30-F	TTTGGAGAGGACACGCTCGAGATGCTGGAAAGTTCTGTTTTGTTG
瞬时表达/过表达载体 Transient expression / overexpression vector	StTrihelix30-R	GCCCTTGCTCACCATGAATTCACATTCTGTTCTTTCTGATGCTTGTT
BD载体 BD carrier	StTrihelix30-F	AGGCCGAATTCCCGGGGATCCTTATGCTGGAAAGTTCTGTTTTGTTG
BD载体 BD carrier	StTrihelix30-R	CCGCTGCAGGTCGACGGATCCTCAACATTCTGTTCTTTCTGATGCT

物种 Species	基因 Gene	氨基酸数量 No. of amino acid	分子量/Da Molecular weight	等电点 pI	脂溶性系数 Aliphatic index	亲水性 Hydropathicity
马铃薯Solanum tuberosum	StTrihelix30	503	57 413.52	7.99	59.96	-1.001
多疣茄Solanum verrucosum	XM_049514945.1	497	56 894.05	7.98	61.09	-0.998
欧白英Solanum dulcamara	XM_055976455.1	502	57 445.28	7.58	58.92	-1.061
辣椒Capsicum annuum	XM_016695686.2	506	57 402.60	8.74	60.20	-1.001
枸杞Lycium barbarum	XM_060327770.1	480	55 030.00	8.31	61.60	-1.008
黄瓜Cucumis sativus	XM_004147307.3	499	57 254.69	5.55	61.04	-1.097
甜瓜Cucumis melo	XM_008462691.3	500	57 063.60	5.62	61.72	-1.061
牵牛Ipomoea nil	XM_019321946.1	480	54 300.42	6.00	58.42	-1.037

物种 Species	基因 Gene	氨基酸数量 No. of amino acid	分子量/Da Molecular weight	等电点 pI	脂溶性系数 Aliphatic index	亲水性 Hydropathicity
马铃薯Solanum tuberosum	StTrihelix30	503	57 413.52	7.99	59.96	-1.001
多疣茄Solanum verrucosum	XM_049514945.1	497	56 894.05	7.98	61.09	-0.998
欧白英Solanum dulcamara	XM_055976455.1	502	57 445.28	7.58	58.92	-1.061
辣椒Capsicum annuum	XM_016695686.2	506	57 402.60	8.74	60.20	-1.001
枸杞Lycium barbarum	XM_060327770.1	480	55 030.00	8.31	61.60	-1.008
黄瓜Cucumis sativus	XM_004147307.3	499	57 254.69	5.55	61.04	-1.097
甜瓜Cucumis melo	XM_008462691.3	500	57 063.60	5.62	61.72	-1.061
牵牛Ipomoea nil	XM_019321946.1	480	54 300.42	6.00	58.42	-1.037

物种 Species	基因 Gene	a-螺旋/% Alpha helix	β-折叠/% Beta turn	不规则卷曲/% Random coil	延伸链/% Extended strand	亚细胞定位 Subcellular localization
马铃薯Solanum tuberosum	XM_006350958.2	43.94	4.57	48.51	2.98	细胞核Nuclear
多疣茄Solanum verrucosum	XM_049514945.1	43.66	4.63	48.89	2.82	细胞核、叶绿体Nuclear、Chloroplast
欧白英Solanum dulcamara	XM_055976455.1	45.22	4.98	48.41	1.39	细胞核Nuclear
辣椒Capsicum annuum	XM_016695686.2	43.87	4.15	50.00	1.98	细胞核Nuclear
枸杞Lycium barbarum	XM_060327770.1	46.04	3.75	48.54	1.67	细胞核Nuclear
黄瓜Cucumis sativus	XM_004147307.3	47.90	3.61	45.29	3.21	细胞核Nuclear
甜瓜Cucumis melo	XM_008462691.3	47.00	4.60	45.60	2.80	细胞核Nuclear
牵牛Ipomoea nil	XM_019321946.1	46.66	3.75	47.71	2.08	细胞核Nuclear
马铃薯Solanum tuberosum	StTrihelix30	43.94	4.57	48.51	2.98	细胞核Nuclear

Cloning of the StTrihelix30 Gene in Potatoes and Functional Analysis Under Low-temperature Stress

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