Welcome to Acta Agronomica Sinica，

Acta Agronomica Sinica

   

Cloning of the IbOPR2 gene promoter and identification of regulatory factors in sweetpotato

Wang Yi-Han1,2,Li Fu-Chang1, Liu Yi1,2,*,Zhu Guo-Peng1,2,*   

  1. 1 School of Breeding and Multiplication / Sanya Institute of Breeding and Multiplication, Hainan University, Sanya 572025, Hainan, China; 2 Key Laboratory for Quality Regulation of Tropical Horticultural Crops of Hainan Province / Tropical Agriculture and forestry College, Hainan University, Haikou 570228, Hainan, China
  • Received:2025-07-02 Revised:2025-11-19 Accepted:2025-11-19 Published:2025-11-26
  • Contact: 刘意, E-mail: 996178@hainanu.edu.cn; 朱国鹏, E-mail: zhuguopeng@hainanu.edu.cn E-mail:23210902000002@hainanu.edu.cn
  • Supported by:
    This study was supported by the “Yazhou Bay” Elite Talent Project Fund (SKJC-JYRC-2024-22) and the China Agriculture Research System of MOF and MARA (CARS-10-Sweetpotato).

PDF

1

Abstract: Jasmonic acids (JAs), as endogenous plant hormones, play a crucial role in the plant response to salt stress. 12-oxophytodienoate reductase (OPR), a key enzyme in JA biosynthesis, remains poorly understood in terms of its expression regulation. To identify transcription factors involved in regulating IbOPR2 expression, this study used the salt-tolerant sweetpotato cultivar ‘Haida 7791’ to clone the promoter sequence of the IbOPR2 gene. Bioinformatics analysis revealed various cis-acting elements, and transcriptional activation assays confirmed promoter activity. Yeast one-hybrid (Y1H) screening was conducted to identify transcription factors interacting with the promoter, and point-to-point Y1H assays were used to validate specific binding. A 1964 bp promoter sequence upstream of IbOPR2 was successfully cloned, containing recognition motifs such as MYBHv1, MYB, and WRKY, along with stress-responsive elements (MYB, MYC) and MeJA-responsive elements, indicating strong transcriptional activation potential. Five candidate transcription factors—PHL7, ZFP16, SKIP11, TGA2, and ERF2—were identified using MYC and MeJA response elements as bait in Y1H assays. Follow-up point-to-point verification confirmed that IbPHL7, IbZFP16, and IbSKIP11 specifically bind to the IbOPR2 promoter. Dual-luciferase reporter assays further demonstrated that these three transcription factors could activate the IbOPR2 promoter. Moreover, qRT-PCR analysis showed that the expression of IbPHL7, IbZFP16, and IbSKIP11 is responsive to salt stress in sweetpotato. Taken together, these results suggest that these transcription factors may function as upstream regulators of IbOPR2, modulating its expression under salt stress conditions. This study represents the first identification of transcriptional regulators of sweetpotato IbOPR2, providing a foundation for further investigation into its regulatory mechanisms in response to salt stress.

Key words: sweetpotato, IbOPR2, yeast one-hybrid, transcription factor, promoter

[1] 曹清河, 王洁, 戴习彬, 等. 甘薯茎叶研究与利用进展. 西南大学学报(自然科学版), 2023, 45(10): 2–10.

Cao Q H, Wang J, Dai X B, et al. Progress on research and utilization of sweetpotato vines and leaves. J Southwest Univ Nat Sci Ed, 2023, 45(10): 2–10 (in Chinese with English abstract).

[2] 宫庆友, 陈冰琳, 李云峰, 等. 盐胁迫对甜玉米幼苗生长的影响. 耕作与栽培, 2021, 41(2): 16–18.
Gong Q Y, Chen B L, Li Y F, et al. Effects of salt stress on sweet corn seedling growth. Tillage Cultiv, 2021, 41(2): 16–18 (in Chinese with English abstract).

[3] 渠晓霞, 黄振英. 盐生植物种子萌发对环境的适应对策. 生态学报, 2005, 25: 2389–2398.
Qu X X, Huang Z Y. The adaptive strategies of halophyte seed germination. Acta Ecol Sin, 2005, 25: 2389–2398 (in Chinese with English abstract).

[4] Li J G, Pu L J, Han M F, et al. Soil salinization research in China: Advances and prospects. J Geogr Sci, 2014, 24: 943–960.

[5] 杨文涛, 王琦, 郭二丹, 等. 土壤盐渍化对植物影响的研究进展. 农业与技术, 2024, 44(18): 95–99.
Yang W T, Wang Q, Guo E D, et al. Research progress on the influence of soil salinization on plants. Agric Technol, 2024, 44(18): 95–99 (in Chinese with English abstract).

[6] 顾鹏鹏, 马鑫磊, 姚锐, 等. 谷子HSP90基因家族鉴定及干旱胁迫下表达分析. 江苏农业科学, 2022, 50(6): 45–52.
Gu P P, Ma X L, Yao R, et al. Identification and expression analysis of HSP90 genes family in foxtail millet (Setaria italica L.) under drought stress. Jiangsu Agric Sci, 2022, 50(6): 45–52 (in Chinese with English abstract).

[7] Howe G A, Major I T, Koo A J. Modularity in jasmonate signaling for multistress resilience. Annu Rev Plant Biol, 2018, 69: 387–415.

[8] Han G Z. Evolution of jasmonate biosynthesis and signaling mechanisms. J Exp Bot, 2017, 68: 1323–1331.

[9] Yastreb T O, Kolupaev Y E, Shkliarevskyi M A, et al. Participation of jasmonate signaling components in the development of Arabidopsis thaliana’s salt resistance induced by H2S and NO donors. Russ J Plant Physiol, 2020, 67: 827–834.

[10] 连青龙, 辛海波, 李晓昕, 等. 异源表达唐菖蒲GhOPR3提高了拟南芥的抗逆性. 园艺学报, 2014, 41: 498–508.
Lian Q L, Xin H B, Li X X, et al. Heterologous expression of Gladiolus GhOPR3 enhances the abiotic stress resistance of Arabidopsis. Acta Hortic Sin, 2014, 41: 498–508 (in Chinese with English abstract).

[11] Dong W, Wang M C, Xu F, et al. Wheat oxophytodienoate reductase gene TaOPR1 confers salinity tolerance via enhancement of abscisic acid signaling and reactive oxygen species scavenging. Plant Physiol, 2013, 161: 1217–1228.

[12] Wang Y K, Yuan G L, Yuan S H, et al. TaOPR2 encodes a 12-oxo-phytodienoic acid reductase involved in the biosynthesis of jasmonic acid in wheat (Triticum aestivum L.). Biochem Biophys Res Commun, 2016, 470: 233–238.

[13] Guang Y, Luo S, Ahammed G J, et al. The OPR gene family in watermelon: genome-wide identification and expression profiling under hormone treatments and root-knot nematode infection. Plant Biol, 2021, 23: 80–88.

[14] Gao L T, Jia S Z, Cao L, et al. An F-box protein from wheat, TaFBA-2A, negatively regulates JA biosynthesis and confers improved salt tolerance and increased JA responsiveness to transgenic rice plants. Plant Physiol Biochem, 2022, 182: 227–239.

[15] Hu Q, Zhu L F, Zhang X N, et al. GhCPK33 negatively regulates defense against Verticillium dahliae by phosphorylating GhOPR3. Plant Physiol, 2018, 178: 876–889.

[16] 王建伟, 许光龄, 陈艳丽, 等. 外源茉莉酸甲酯对盐胁迫下菜用甘薯生长生理的影响. 中国瓜菜, 2022, 35(6): 69–75.
Wang J W, Xu G L, Chen Y L, et al. Exogenous methyl jasmonate affects growth and physiology of leafy sweet potato under salt stress. China Cucurbits Veg, 2022, 35(6): 69–75 (in Chinese with English abstract).

[17] Li W X, Li Y P, Xu Y, et al. Genome-wide identification, gene cloning, subcellular location and expression analysis of the OPR gene family under salt stress in sweetpotato. BMC Plant Biol, 2024, 24: 1171.

[18] 黄婷. 菜用甘薯耐盐品种筛选及耐盐生理响应. 海南大学硕士学位论文, 海南海口, 2020.

Huang T. Screening of Salt-tolerant Varieties of Vegetable Sweet Potato and Salt-tolerant Physiological Response. MS Thesis of Hainan University, Haikou, Hainan, China, 2020 (in Chinese with English abstract).

[19] Barragán-Rosillo A C, Peralta-Alvarez C A, Ojeda-Rivera J O, et al. Genome accessibility dynamics in response to phosphate limitation is controlled by the PHR1 family of transcription factors in Arabidopsis. Proc Natl Acad Sci USA, 2021, 118: e2107558118.

[20] Sun L C, Song L, Zhang Y, et al. Arabidopsis PHL2 and PHR1 act redundantly as the key components of the central regulatory system controlling transcriptional responses to phosphate starvation. Plant Physiol, 2016, 170: 499–514.

[21] He K R, Du J C, Han X, et al. Phosphate starvation response1 (phr1) interacts with jasmonate zim-domain (jaz) and MYC2 to modulate phosphate deficiency-induced jasmonate signaling in Arabidopsis. Plant Cell, 2023, 35: 2132–2156.

[22] Yang W T, Bae K D, Lee S W, et al. The myb-cc transcription factor phosphate starvation response-like 7 (phl7) functions in phosphate homeostasis and affects salt stress tolerance in rice. Plants, 2024, 13: 637.

[23] Han G L, Lu C X, Guo J R, et al. C2H2 zinc finger proteins: master regulators of abiotic stress responses in plants. Front Plant Sci, 2020, 11: 115.

[24] Huang J, Yang X, Wang M M, et al. A novel rice C2H2-type zinc finger protein lacking DLN-box/EAR-motif plays a role in salt tolerance. Biochim Biophys Acta, 2007, 1769: 220–227.

[25] Zhang X, Guo X P, Lei C L, et al. Overexpression of SlCZFP1, a novel TFIIIA-type zinc finger protein from tomato, confers enhanced cold tolerance in transgenic Arabidopsis and rice. Plant Mol Biol Rep, 2011, 29: 185–196.

[26] Dinkins R D, Tavva V S, Palli S R, et al. Mutant and overexpression analysis of a C2H2 single zinc finger gene of Arabidopsis. Plant Mol Biol Re, 2012, 30: 99–110.

[27] Wang F B, Tong W J, Zhu H, et al. A novel Cys2/His2 zinc finger protein gene from sweetpotato, IbZFP1, is involved in salt and drought tolerance in transgenic Arabidopsis. Planta, 2016, 243: 783–797.

[28] Zhang Z Y, Liu H H, Sun C, et al. A C2H2 zinc-finger protein OsZFP213 interacts with OsMAPK3 to enhance salt tolerance in rice. J Plant Physiol, 2018, 229: 100–110.

[29] 王洪云, 黄剑, 赖钊, 等. 植物F-box蛋白质及其研究进展. 科学通报, 2002, 47: 891–895.
Wang H Y, Huang J, Lai Z, et al. Plant F-box protein and its research progress. Chin Sci Bull, 2002, 47: 891–895 (in Chinese with English abstract).

[30] 李莉, 李懿星, 夏凯, 等. 植物F-Box蛋白及其生物学功能研究. 安徽农业科学, 2010, 38: 19879–19881.
Li L, Li Y X, Xia K, et al. Plant F-box proteins and their biological functions. J Anhui Agric Sci, 2010, 38: 19879–19881 (in Chinese with English abstract).

[31] Hassan M N U, Ismail I. Evaluation of genetic and metabolic role of SKIP11 in Arabidopsis thaliana. AIP Conf Proc, 2015, 1678: 030034.

[1] MA Juan-E, YAO You-Hua, YAO Xiao-Hua, WU Kun-Lun, CUI Yong-Mei. Cloning and functional analysis of HvERF039 gene in Qingke (Hordeum vulgare L. var. nudum Hook. f) [J]. Acta Agronomica Sinica, 2025, 51(9): 2341-2357.
[2] YANG Hai-Yang, WU Lin-Xuan, LI Bo-Wen, SHI Han-Feng, YUAN Xi-Long, LIU Jin-Zhao, CAI Hai-Rong, CHEN Shi-Yi, GUO Tao, WANG Hui. OsWRI3, identified based on QTL mapping, regulates seed shattering in rice [J]. Acta Agronomica Sinica, 2025, 51(7): 1712-1724.
[3] YIN Yu-Meng, WANG Yan-Nan, KANG Zhi-He, QIAO Shou-Chen, BIAN Qian-Qian, LI Ya-Wei, CAO Guo-Zheng, ZHAO Guo-Rui, XU Dan-Dan, YANG Yu-Feng. Cloning and functional analysis of glutathione S-transferase gene IbGSTU7 in sweetpotato [J]. Acta Agronomica Sinica, 2025, 51(7): 1736-1746.
[4] SHEN Ao, LIU Min, NI Di-An, LIU Wei. Promoter characterization and expression pattern analysis of the m6A methyltransferase gene SiMTA1 in foxtail millet [J]. Acta Agronomica Sinica, 2025, 51(7): 1969-1978.
[5] HUANG Meng-Xin, ZHUANG Ling-Ling, CHENG Pei-Pei, LI Qin, XU Jian-Tang, TAO Ai-Fen, FANG Ping-Ping, QI Jian-Min, ZHANG Li-Wu. Cloning and transcriptional activity analysis of U6 promoter in jute (Corchorus capsularis) [J]. Acta Agronomica Sinica, 2025, 51(5): 1156-1165.
[6] FANG Ying-Hao, ZHOU Bo, CHEN Ru-Mei, YANG Wen-Zhu, QIN Hui-Min. Integrative analysis of RNA-seq and PER-seq to elucidate regulatory network of ZmHDZ6 expression [J]. Acta Agronomica Sinica, 2025, 51(4): 958-968.
[7] PAN Ju-Zhong, WEI Ping, ZHU De-Ping, SHAO Sheng-Xue, CHEN Shan-Shan, WEI Ya-Qian, GAO Wei-Wei. Cloning and functional analysis of OsERF104 transcription factor in rice [J]. Acta Agronomica Sinica, 2025, 51(4): 900-913.
[8] WANG Lin, CHEN Xiao-Yu, ZHANG Wen-Meng-Long, WANG Si-Qi, CHENG Bing-Yun, CHENG Jing-Qiu, PAN Rui, ZHANG Wen-Ying. Molecular characteristics and functional analysis of HvMYB2 in response to drought stress in barley [J]. Acta Agronomica Sinica, 2025, 51(4): 873-887.
[9] HUO Ru-Xue, GE Xiang-Han, SHI Jia, LI Xue-Rui, DAI Sheng-Jie, LIU Zhen-Ning, LI Zong-Yun. Functional analysis of the sweetpotato histidine kinase protein IbHK5 in response to drought and salt stresses [J]. Acta Agronomica Sinica, 2025, 51(3): 650-666.
[10] WANG Yu-Xin, CHEN Tian-Yu, ZHAI Hong, ZHANG Huan, GAO Shao-Pei, HE Shao-Zhen, ZHAO Ning, LIU Qing-Chang. Cloning and characterization of drought tolerance function of kinase gene IbHT1 in sweetpotato [J]. Acta Agronomica Sinica, 2025, 51(2): 301-311.
[11] AI Li, LI Meng-Tao, LU Fa-Bao, LIU Xiao-Chen, MAI Wei-Tao, ZHOU Xin-Cheng, CHEN Xin. Molecular mechanism of the MeERF6-MePAP2 module in response to low-temperature stress [J]. Acta Agronomica Sinica, 2025, 51(11): 2971-2982.
[12] LI Wan, CHANG Zi-Rui, LU Yao, SHEN Ri-Min, ZHAO Yong-Ping, BAI Xiao-Dong. Identification of RAV family in 25 different plant species and expression analysis of RAV genes in potato [J]. Acta Agronomica Sinica, 2025, 51(11): 2944-2957.
[13] ZHANG Shun-Jie, WU Wei-Tai, RAN Xi-Yue, ZHAO Zi-Han, HAN Yong-Hui, WU Zheng-Dan, ZHANG Kai. Functional analysis of the sweetpotato β-Amylase IbBAM48829 [J]. Acta Agronomica Sinica, 2025, 51(11): 3096-3104.
[14] SONG Qian-Na, SONG Hui-Yang, LI Jing-Hao, DUAN Yong-Hong, MEI Chao, FENG Rui-Yun. Response of transcription factor StFBH3 under abiotic stress in potato [J]. Acta Agronomica Sinica, 2025, 51(1): 247-259.
[15] GUO Fei-Xiang, LI Chun-Xia, ZHOU Shuang, GUO Bin-Bin, ZHANG Jun, MA Chao. Identification of the R2R3-MYB transcription factor family and screening of genes regulating flavonoid synthesis in mung bean [J]. Acta Agronomica Sinica, 2025, 51(1): 117-133.
Viewed
Full text


Abstract

Cited
  Shared   
  Discussed   
No Suggested Reading articles found!
Add: No. 80 Xueyuan South Rd, Beijing 100081, P. R. China E-mail: zwxb301@caas.cn
Supported by Beijing Magtech Co.Ltd