Please wait a minute...
Journal of Integrative Agriculture  2014, Vol. 13 Issue (6): 1230-1238    DOI: 10.1016/S2095-3119(13)60572-9
Crop Genetics · Breeding · Germplasm Resources Advanced Online Publication | Current Issue | Archive | Adv Search |
Cloning, Localization and Expression Analysis of ZmHsf-like Gene in Zea mays
 LI Hui-cong, LI Guo-liang, LIU Zi-hui, ZHANG Hong-mei, ZHANG Yan-min , GUO Xiu-lin
Institute of Genetics and Physiology, Hebei Academy of Agriculture and Forestry Sciences/Plant Genetic Engineering Center of Hebei Province, Shijiazhuang 050051, P.R.China
Download:  PDF in ScienceDirect  
Export:  BibTeX | EndNote (RIS)      
摘要  Using homology cloning method, a heat shock transcription factor (Hsf) like gene, ZmHsf-like, was cloned from maize (Zea mays) leaves. Sequence analyses showed that the open reading frame (ORF) of the gene ZmHsf-like is 1404 bp long, encoding 467 amino acids. The sequence of amino acids encoded by ZmHsf-like contains the most conserved and typical DNA-binding domain of Hsf family. By bombardment into onion epidermis, we firstly found that the ZmHsf-like was subcellular-located in nucleus. NucPred analysis revealed there is a classic NLS of KKRR peptide in protein. Real-time PCR showed that ZmHsf-like gene expressed in leaves, stems and roots of maize seedlings under normal growth conditions, and the highest expression level was in roots, lower in leaves and the lowest in stems. The ZmHsf-like gene expression could be up-regulated by heat shock, PEG, ABA, and H2O2 in different degrees, among which the heat shock and ABA worked more efficiently. Obvious differences of the peak value and its corresponding time point of ZmHsf-like gene expression were observed among treatments. Experiments with inhibitor further suggested that the up-regulation ZmHsf-like gene expression of heat shock was H2O2-dependent while the induction of ZmHsf-like with PEG did not depend on the existence of H2O2. These results pointed out that ZmHsf-like gene probably regulates responsive reactions to abiotic stresses especially heat shock and drought through different signal transduction pathways.

Abstract  Using homology cloning method, a heat shock transcription factor (Hsf) like gene, ZmHsf-like, was cloned from maize (Zea mays) leaves. Sequence analyses showed that the open reading frame (ORF) of the gene ZmHsf-like is 1404 bp long, encoding 467 amino acids. The sequence of amino acids encoded by ZmHsf-like contains the most conserved and typical DNA-binding domain of Hsf family. By bombardment into onion epidermis, we firstly found that the ZmHsf-like was subcellular-located in nucleus. NucPred analysis revealed there is a classic NLS of KKRR peptide in protein. Real-time PCR showed that ZmHsf-like gene expressed in leaves, stems and roots of maize seedlings under normal growth conditions, and the highest expression level was in roots, lower in leaves and the lowest in stems. The ZmHsf-like gene expression could be up-regulated by heat shock, PEG, ABA, and H2O2 in different degrees, among which the heat shock and ABA worked more efficiently. Obvious differences of the peak value and its corresponding time point of ZmHsf-like gene expression were observed among treatments. Experiments with inhibitor further suggested that the up-regulation ZmHsf-like gene expression of heat shock was H2O2-dependent while the induction of ZmHsf-like with PEG did not depend on the existence of H2O2. These results pointed out that ZmHsf-like gene probably regulates responsive reactions to abiotic stresses especially heat shock and drought through different signal transduction pathways.
Keywords:  maize       Hsf       ZmHsf-like       stress       nuclear-localization  
Received: 03 April 2013   Accepted:
Fund: 

This work was supported by grants from the Key Project of Application Basic Research of Hebei Province, China (12965517D) and the Youth Fund of Hebei Academy of Agriculture and Forestry Sciences, China (A09110103).

Corresponding Authors:  GUO Xiu-lin, Tel/Fax: +86-311-87269032, E-mail: myhf2002@163.com   
About author:  LI Hui-cong, E-mail: lhc_009@163.com

Cite this article: 

LI Hui-cong, LI Guo-liang, LIU Zi-hui, ZHANG Hong-mei, ZHANG Yan-min , GUO Xiu-lin. 2014. Cloning, Localization and Expression Analysis of ZmHsf-like Gene in Zea mays. Journal of Integrative Agriculture, 13(6): 1230-1238.

Agashe V R, Hartl F U. 2000. Roles of molecular chaperones in cytoplasmic protein folding. Seminars in Cell & Developmental Biology, 11, 15-25

 Almoguera C, Rojas A, Díaz-Martín J, Prieto-Dapena P, Carranco R, Jordano J 2002. A seed-specific heat-shock transcription factor involved in developmental regulation during embryogenesis in sunflower. The Journal of Biological Chemistry, 277, 43866-43872

 Aranda M A, Escaler M, Thomas C L, Maule A J 1999. A heat shock transcription factor in pea is differentially controlled by heat and virus replication. The Plant Journal, 20, 153-161

 Brameier M, Krings A, MacCallum R M 2007. NucPred-predicting nuclear localization of proteins. Bioinformatics Applications Note, 23, 1159-1160

Busch W, Wunderlich M, Schöffl F 2005. Identification of novel heat shock factor-dependent genes and biochemical pathways in Arabidopsis thaliana. The Plant Journal, 14, 1-14

 Charng Y Y, Liu H C, Liu N Y, Chi W T, Wang C N, Chang S H, Wang T T. 2007. A heat-induced transcription factor, HsfA2, is required for extension of acquired thermotolerance in Arabidopsis. Plant Physiology, 143, 251-262

 Czarnecha-Verner, Yuan C X, Fox P C, Gurley W B. 1995. Isolation and characterization of six heat shock transcription factor cDNA clones from soybean. Plant Molecular Biology, 29, 37-51

 Dang J, Jiang M Y, Lin F. 2010. ABA up-regulate the expression of OsHsf gene in leaves of rice plants. Journal of Nanjing Agricultural University, 33, 11-15 (in Chinese)

Dat J F, Foyer C H, Scott I M. 1998. Changes in salicylic acid and antioxidants during induction of thermotolerance in mustard seedlings. Plant Physiology, 118, 1455-1461

Davletova S, Rizhsky L, Liang H J, Zhong S Q, David J O, Jesse C, Vladimir S, Karen S, Ron M. 2005. Cytosolic ascorbate peroxidase1 is a central component of the reactive oxygen gene network of Arabidopsis. The Plant Cell, 17, 268-281

 Ellis R J. 2000. Chaperone substrates inside the cell. Trends in Biochemical Science, 25, 210-212

 Gagiardi D, Breton C, Chaboud A, Vergne P, Dumas C. 1995. Expression of heat shock factor and heat shock protein 70 genes during maize pollen development. Plant Molecular Biology, 29, 841-856

 Guo J K, Wu J, Ji Q, Wang C, Luo L, Yuan Y, Wang Y, Wang J. 2008. Genome-wide analysis of heat shock transcription factor families in rice and Arabidopsis. Journal of Genetics and Genomics, 35, 105-118

 Heerklotz D, Döring P, Bonzelius F, Winkelhaus S, Nover L. 2001. The balance of nuclear import and export determines the intrancellular distribution and function of tomato heat stress transcription factor HsfA2. Molecular and Cellular Biology, 21, 1759-1768

 Hübel A, Schöffl F. 1994. Arabidopsis heat shock factor: isolation and characterization of the gene and the recombinant protein. Plant Molecular Biology, 26, 353- 362.

Jolly C, Morimoto R I. 2000. Role of the heat shock response and molecular chaperones in oncogenesis and cell death. Journal of the National Cancer Iinstitute, 92, 1564-1572

 Lee B H, Won S H, Lee H S, Miyao M, Chung W I, Kim I J. 2000. Expression of the chloroplast-localized small heat shock protein by oxidative stress in rice. Gene, 245, 283-290

 Lee J H, Hübel A, Schöffl F. 1995. Derepression of the activity of genetically engineered heat shock factor causes constitutive synthesis of heat shock proteins and increased thermotolerance in transgenic Arabidopsis. The Plant Journal, 8, 603-612

 Li C G, Chen Q J, Gao X Q, Qi B S, Cheng N Z, Xu S M, Chen J, Wang X C. 2005. Heat shock transcription factor AtHsfA2 regulating genes expression related to stresses and increase endurance to heat and oxidation stress in Arabidopsis. Chinese Science (C-Life Science), 35, 398- 407.

Lin Y X, Jiang H Y, Chu Z X, Tang X L, Zhu S W, Cheng B J. 2011. Genome-wide identification, classification and analysis of heat shock transcription factor family in maize. BMC Genomics, 12, 76-89

 Liu H C, Liao H T, Charng Y Y. 2011. The role of class A1 heat shock factors (HSFA1s) in response to heat and other stresses in Arabidopsis. Plant Cell and Environment, 34, 738-751

 Lohmann C, Eggers-Schumacher G, Wunderlich M, Schöffl F. 2004. Two different heat shock transcription factors regulate immediate early expression of stress genes in Arabidopsis. Molecular Genetics and Genomics, 271, 11-21

 Mishra S K, Tripp J, Winkelhaus S, Tschiersch B, Theres K, Nover L, Scharf K D. 2002. In the complex family of heat stress transcription factors, HsfA1 has a unique role as master regulator of thermotolerance in tomato. Genes and Development, 16, 1555-1567

 Nishizawa A, Yabuta Y, Yoshida E, Maruta T, Yoshimura K, Shigeoka S. 2006. Arabidopsis heat shock transcription factor A2 as a key regulator in response to several types of environmental stress. The Plant Journal, 48, 535-547

 Nishizawa-Yokoi A, Nosaka R, Hayashi H, Tainaka1 H, Maruta1 T, Tamoi M, Ikeda M, Ohme-Takagi M, Yoshimura K, Yabuta Y, Shigeoka S. 2011. HsfA1d and HsfA1e involved in the transcriptional regulation of HsfA2 function as key regulators for the Hsf signaling network in response to environmental stress. Plant Cell Physiology, 52, 933-945

 Nover L, Bharti K, Döring P, Mishra S K, Ganguli A, Scharf K D. 2001. Arabidopsis and the heat stress transcription factor world: How many heat stress transcription factors do we need. Cell Stress Chaperones, 6, 177-189

 Nover L, Scharf K D, Gagliardi D, Vergne P, Czarnecka- Verner E, Gurley W B. 1996. The Hsf world: Classification and properties of plant heat stress transcription factors. Cell Stress Chaperones, 1, 215-223

 Scharf K D, Heider H, Höhfeld I, Lyck R, Schmidt E, Nover L. 1998. The tomato Hsf system: HsfA2 needs interaction with HsfA1 for efficient nuclear import and may be localized in cytoplasmic heat stress granules. Molecular and Cellular Biology, 18, 2240-2251

 Scharf K D, Rose S, Zott W, Schöffl F, Nover L. 1990. Three tomato genes code for heat stress transcription factors with a region of remarkable homology to the DNA-binding domain of the yeast HSF. EMBO Journal, 9, 4495-4501

 Schett G, Steiner C W, Groger M, Winkler S, Graninger W, Smolen J. 1999. Activation of Fas inhibits heat induced activation of Hsf1 and upregulation of Hsp70. Federation of American Societies for Experimental Biology Journal, 13, 833-842

 Schramm F, Ganguli A, Kiehlmann E, Englich G, Walch D, von Kaskull-Doring P. 2006. The heat stress transcription factor HsfA2 serves as a regulatory amplifier of a subset of genes in the heat stress response in Arabidopsis. Plant Molecular Biology, 60, 759-772

 Schöffl F, Prändl R, Reindl A. 1998. Regulation of the heat- shock response. Plant Physiology, 117, 1135-1141

 Shim D, Hwang J U, Lee J, Lee S, Choi Y, An G, Martinoia E, Lee Y. 2009. Orthologs of the class A4 heat shock transcription factor HsfA4a confer cadmium tolerance in wheat and rice. The Plant Journal, 21, 4031-4043

 Yamanouchi U, Yano M, Lin H X, Ashikari M, Yamada K. 2002. Rice spotted leaf gene, Spl7, encodes a heat stress transcription factor protein. Proceedings of the National Academy of Sciences of the United States of America, 99, 7530-7535

 Yokotani N, Ichikawa T, Kondou Y, Matsui M, Hirochika H, Iwabuchi M, Oda K. 2008. Expression of rice heat stress transcription factor OsHsfA2e enhances tolerance to environmental stresses in transgenic Arabidopsis. Planta, 227, 957-967
[1] Teng Li, Shumei Wang, Qing Liu, Xuepeng Zhang, Lin Chen, Yuanquan Chen, Wangsheng Gao, Peng Sui. Effects of changing assimilate supply on starch synthesis in maize kernels under high temperature stress[J]. >Journal of Integrative Agriculture, 2026, 25(2): 639-647.
[2] Zhuandi Wu, Xin Hu, Wenzhi Wang, Zhengying Luo, Naveed ur Rehman, Peifang Zhao, Jiayong Liu, Shuzhen Zhang, Fenggang Zan, Xinlong Liu, Jiawen Guo. ScD27.2 gene regulation mechanism during sugarcane tillering and growth[J]. >Journal of Integrative Agriculture, 2026, 25(2): 694-708.
[3] Xiaohui Xu, Qiang Chai, Falong Hu, Wen Yin, Zhilong Fan, Hanting Li, Zhipeng Liu, Qiming Wang. Intercropping grain crops with green manure under reduced chemical nitrogen improves the soil carbon stocks by optimizing aggregates in an oasis irrigation area[J]. >Journal of Integrative Agriculture, 2026, 25(1): 326-338.
[4] Zhilong Hao, Yuping Zhang, Weiyi Kong, Jiao Feng, Yucheng Zheng, Hongzheng Lin, Xiaomin Yu, Yun Sun, Xiangxiang Huang, Wei Wang, Yang Wu, Xinyi Jin. Mechanical stress induces molecular changes in oolong tea: Insights from multi-omics analysis[J]. >Journal of Integrative Agriculture, 2026, 25(1): 352-365.
[5] Shuo Ma, Tong Li, Ziquan Feng, Yali Zhang, Han Jiang, Yuanyuan Li. MdXTH30, an apple gene encoding endotransferase/hydrolase for xyloglucan, enhances plant resistance to drought, salt and pathogenic stresses[J]. >Journal of Integrative Agriculture, 2026, 25(1): 127-137.
[6] Yanqing Wu, Jiao Liu, Lu Zhao, Hao Wu, Yiming Zhu, Irshad Ahmad, Guisheng Zhou. Abiotic stress responses in crop plants: A multi-scale approach[J]. >Journal of Integrative Agriculture, 2026, 25(1): 1-15.
[7] Mohan K. Bista, Purushothaman Ramamoorthy, Ranadheer Reddy Vennam, Sadikshya Poudel, K. Raja Reddy, Raju Bheemanahalli. Impacts of abiotic stresses on cotton physiology and vigor under current and future CO2 levels[J]. >Journal of Integrative Agriculture, 2026, 25(1): 105-117.
[8] Ziwen Shi, Sheng Zhang, Qing He, Xiaoyuan Wang, Bo yang, Tao Yu, Hongyang Yi, Tingzhao Rong, Moju Cao. ZmCals12 impacts maize growth and development by regulating symplastic transport[J]. >Journal of Integrative Agriculture, 2026, 25(1): 42-55.
[9] Qinghao Wang, Juan Hu, Weizhen Yu, Limin Gu, Peng Liu, Bin Zhao, Wenchao Zhen, Jiwang Zhang, Baizhao Ren. Shading and waterlogging interactions exacerbate summer maize yield losses by reducing assimilate accumulation and remobilization processes[J]. >Journal of Integrative Agriculture, 2026, 25(1): 92-104.
[10] Lichao Zhai, Shijia Song, Lihua Zhang, Jinan Huang, Lihua Lv, Zhiqiang Dong, Yongzeng Cui, Mengjing Zheng, Wanbin Hou, Jingting Zhang, Yanrong Yao, Yanhong Cui, Xiuling Jia. Subsoiling before winter wheat alleviates the kernel position effect of densely grown summer maize by delaying post-silking root–shoot senescence[J]. >Journal of Integrative Agriculture, 2025, 24(9): 3384-3402.
[11] Ling Ai, Ju Qiu, Jiuguang Wang, Mengya Qian, Tingting Liu, Wan Cao, Fangyu Xing, Hameed Gul, Yingyi Zhang, Xiangling Gong, Jing Li, Hong Duan, Qianlin Xiao, Zhizhai Liu. A naturally occurring 31 bp deletion in TEOSINTE BRANCHED1 causes branched ears in maize[J]. >Journal of Integrative Agriculture, 2025, 24(9): 3322-3333.
[12] Jing Zhou, Bingshuai Du, Yibo Cao, Kui Liu, Zhihua Ye, Yiming Huang, Lingyun Zhang. Genome-wide identification of sucrose transporter genes in Camellia oleifera and characterization of CoSUT4[J]. >Journal of Integrative Agriculture, 2025, 24(9): 3494-3510.
[13] Siya Li, Lu Cao, Ziwen Zhou, Yaohua Cheng, Xianchen Zhang, Yeyun Li. The miR164a targets CsNAC1 to negatively regulate the cold tolerance of tea plants (Camellia sinensis)[J]. >Journal of Integrative Agriculture, 2025, 24(8): 3073-3086.
[14] Jie Zhang, Han Gao, Fuhao Ren, Zehua Zhou, Huan Wu, Huahua Zhao, Lu Zhang, Mingguo Zhou, Yabing Duan. The stress regulator FgWhi2 and phosphatase FgPsr1 play crucial roles in the regulation of secondary metabolite biosynthesis and the response to fungicides in Fusarium graminearum[J]. >Journal of Integrative Agriculture, 2025, 24(8): 3095-3111.
[15] Xuehao Zhang, Qiuling Zheng, Yongjiang Hao, Yingying Zhang, Weijie Gu, Zhihao Deng, Penghui Zhou, Yulin Fang, Keqin Chen, Kekun Zhang. Physiology and transcriptome profiling reveal the drought tolerance of five grape varieties under high temperatures[J]. >Journal of Integrative Agriculture, 2025, 24(8): 3055-3072.
No Suggested Reading articles found!