Acta Physiologiae Plantarum

, Volume 35, Issue 2, pp 575–587 | Cite as

Transcriptional profile of the spring freeze response in the leaves of bread wheat (Triticum aestivum L.)

  • Guozhang Kang
  • Gezi Li
  • Wenping Yang
  • Qiaoxia Han
  • Hongzhen Ma
  • Yonghua Wang
  • Jiangping Ren
  • Yunji Zhu
  • Tiancai Guo
Original Paper


Measurement of the electrolyte leakage rates in wheat leaves indicated that there was no significant difference in susceptibility to −5 °C spring freeze stress among five bread wheat cultivars at the floret primordium-differentiating stage of spike development. A global transcriptional profile was created using the Affymetrix Wheat GeneChip microarray for one wheat cultivar (Yumai 34) under −5 °C freeze stress. After assaying genes with significant regulation at 1 and 3 days after −5 °C freeze stress, we identified 600 genes that were previously annotated as showing changes in expression of at least than two-fold at one or both of the time points. Among these genes, we further analysed 102 genes whose expression levels changed at least eight-fold after 1 or 3 days of freeze stress. These genes encoded an ice recrystallization protein, cold-related proteins, CBF transcription factors, calcium-dependent protein kinases, Na+/H+ antiporters, aquaporins, and many metabolic enzymes. The results of this study were compared with those of a previous study on the sub-freeze hardening response in wheat and spring freeze stress in wheat and barley. Many genes, including those encoding WCOR413, LEA, glycine-rich RNA-binding protein, ferritin, aquaporin 2, and a pathogen-induced protein, showed similar expression levels in these studies. Spring freeze stress is a complex phenomenon involving physiological mechanisms and multiple genes that had not been previously characterised.


cDNA arrays Gene expression Spring freeze stress Triticum aestivum L. 



This study was supported by funds from the National Transgenic Major Project (2009ZX08002-21B), the Special Fund for Agro-scientific Research in the Public Interest (201003002), and the National Basic Research Program of China (2009CB118602).

Conflict of interest

The authors declare no conflict of interest.

Supplementary material

11738_2012_1099_MOESM1_ESM.doc (1.5 mb)
Supplementary material 1 (DOC 1558 kb)


  1. Berrocal-Lobo M, Molina A, Solano R (2002) Constitutive expression of ethylene-response-factor1 in Arabidopsis confers resistance to several necrotrophic fungi. Plant J 29:23–32PubMedCrossRefGoogle Scholar
  2. Breton G, Danyluk J, Charron JBF, Sarhan F (2003) Expression profiling and bioinformatic analyses of a novel stress-regulated multispanning transmembrane protein family from cereals and Arabidopsis. Plant Physiol 132:64–74PubMedCrossRefGoogle Scholar
  3. Chan Z, Wang Q, Xu X, Meng X, Qin G, Li B, Tian S (2008) Functions of defense-related proteins and dehydrogenases in resistance response induced by salicylic acid in sweet cherry fruits at different maturity stages. Proteomics 8:4791–4797PubMedCrossRefGoogle Scholar
  4. Chauvin LP, Houde M, Sarhan F (1993) A leaf-specific gene stimulated by light during wheat acclimation to low temperature. Plant Mol Biol 23:255–265PubMedCrossRefGoogle Scholar
  5. Chen TH, Murata N (2002) Enhancement of tolerance of abiotic stress by metabolic engineering of betaines and other compatible solutes. Curr Opin Plant Biol 5:250–257PubMedCrossRefGoogle Scholar
  6. Chinnusamy V, Zhu J, Zhu J (2006) Gene regulation during cold acclimation in plants. Physiol Plantarum 126:52–61CrossRefGoogle Scholar
  7. Chinnusamy V, Zhu JH, Zhu JK (2007) Cold stress regulation of gene expression in plants. Trends Plant Sci 12:444–451PubMedCrossRefGoogle Scholar
  8. Cook D, Fowler S, Fiehn O, Thomashow MF (2004) A prominent role for the CBF cold response pathway in configuring the low-temperature metabolome of Arabidopsis. Proc Natl Acad Sci USA 101:15243–15248PubMedCrossRefGoogle Scholar
  9. Coupe SA, Watson LM, Ryan DJ, Pinkney TT, Eason JR (2004) Molecular analysis of programmed cell death during senescence in Arabidopsis thaliana and Brassica oleracea: cloning broccoli LSD1, Bax inhibitor and serine palmitoyltransferase homologues. J Exp Bot 55:59–68PubMedCrossRefGoogle Scholar
  10. Crosatti C, Marè C, Mazzucotelli E, Belloni S, Barilli S, Bassi R, Dubcovskyi J, Galiba G, Stanca AM, Cattivelli L (2003) Genetic analysis of the expression of the cold-regulated gene cor14b: a way toward the identification of components of the cold response signal transduction in Triticeae. Can J Bot 81:1162–1167CrossRefGoogle Scholar
  11. Cui JM, Guo TC (2008) Spike of wheat. China Agricultural Press, Beijing (in Chinese)Google Scholar
  12. Danyluk J, Carpentier E, Sarhan F (1996) Identification and characterization of a low temperature regulated gene encoding an actin-binding protein from wheat. FEBS Lett 389:324–327PubMedCrossRefGoogle Scholar
  13. Danyluk J, Perrona A, Houdea M, Liminb A, Fowlerb B, Benhamouc N, Sarhana F (1998) Accumulation of an acidic dehydrin in the vicinity of the plasma membrane during cold acclimation of wheat. Plant Cell 10:623–638PubMedGoogle Scholar
  14. Eulgem T, Somssich IE (2007) Networks of WRKY transcription factors in defense signaling. Curr Opin Plant Biol 10:366–371PubMedCrossRefGoogle Scholar
  15. Foyer CH, Noctor G (2005) Redox homeostasis and antioxidant signaling: a metabolic interface between stress perception and physiological responses. Plant Cell 17:1866–1875PubMedCrossRefGoogle Scholar
  16. Galiba G, Vágújfalvi A, Li C, Soltész A, Dubcovsky J (2009) Regulatory genes involved in the determination of frost tolerance in temperate cereals. Plant Sci 176:12–19CrossRefGoogle Scholar
  17. Gana JA, Sutton F, Kenefick DG (1997) cDNA structure and expression patterns of a low-temperature- specific wheat gene tacr7. Plant Mol Biol 34:643–650PubMedCrossRefGoogle Scholar
  18. Gulick PJ, Drouin S, Yu ZH, Danyluk J, Poisson G, Monroy AF, Sarhan F (2005) Transcriptome comparison of winter and spring wheat responding to low temperature. Genome 48:913–923PubMedCrossRefGoogle Scholar
  19. Hayes PM, Blake T, Chen TTH, Tragoonrung S, Chen F, Pan A, Liu B (1993) Quantitative trait loci on barley (Hordeum vulgare L.) chromosome 7 associated components of winterhardiness. Genome 36:66–71PubMedCrossRefGoogle Scholar
  20. Herman EM, Rotter K, Premakumar R, Elwinger G, Bae R, Ehler-King L, Chen SX, Livingston DP III (2006) Additional freeze hardiness in wheat acquired by exposure to −3 °C is associated with extensive physiological, morphological, and molecular changes. J Exp Bot 57:3601–3618PubMedCrossRefGoogle Scholar
  21. Hrabak EM (2000) Calcium-dependent protein kinases and their relatives. Adv Bot Res 32:185–223CrossRefGoogle Scholar
  22. Hulbert SH, Bai J, Fellers JP, Pacheco MG, Bowden RL (2007) Gene expression patterns in near isogenic lines for wheat rust resistance gene. Phytopathology 97:1083–1093PubMedCrossRefGoogle Scholar
  23. Inada M, Ueda A, Shi W, Takabe T (2005) A stress-inducible plasma membrane protein 3 (AcPMP3) in a monocotyledonous halophyte, Aneurolepidium chinense, regulates cellular Na+ and K+ accumulation under salt stress. Planta 220:395–402PubMedCrossRefGoogle Scholar
  24. Ítámvás P, Saalbach G, Prášil IT, Čapková V, Opatrná J, Ahmed J (2007) WCS120 protein family and proteins soluble upon boiling in cold-acclimated winter wheat. J Plant Physiol 164:1197–1207CrossRefGoogle Scholar
  25. Jian H (2009) From freezing to scorching, transcriptional responses to temperature variations in plants. Curr Opin Plant Biol 12:568–573CrossRefGoogle Scholar
  26. Johnson JD, Gagnon KG (1988) Assessing freeze damage in loblolly pine seedlings: a comparison of ethane production to electrolyte leakage. New Forest 2:65–72CrossRefGoogle Scholar
  27. Kaplan F, Kopka J, Haskell DW, Zhao W, Schiller KC, Gatzke N, Sung DY, Guy CL (2004) Exploring the temperature-stress metabolome of Arabidopsis. Plant Physiol 136:4159–4168PubMedCrossRefGoogle Scholar
  28. Kim JY, Kim WY, Kwak KJ, Oh SH, Han YS, Kang H (2010) Glycine-rich RNA-binding proteins are functionally conserved in Arabidopsis thaliana and Oryza sativa during cold adaptation process. J Exp Bot 61:2317–2325PubMedCrossRefGoogle Scholar
  29. Kobayashi F, Takumi S, Nakata M, Ohno R, Nakamura T, Nakamura C (2004) Comparative study of the expression profiles of the Cor/Lea gene family in two wheat cultivars with contrasting levels of freezing tolerance. Physiol Plantarum 120:585–594CrossRefGoogle Scholar
  30. Kocsy G, Athmer B, Perovic D, Himmelbach A, Szűcs A, Vashegyi I, Schweizer P, Galiba G, Stein N (2010) Regulation of gene expression by chromosome 5A during cold hardening in wheat. Mol Genet Genomics 283:351–363PubMedCrossRefGoogle Scholar
  31. Koo BC, Bushman BS, Mott IW (2008) Transcripts associated with non-acclimated freezing response in two barley cultivars. Plant Genome 1:21–32CrossRefGoogle Scholar
  32. Kosová K, Vítámvás P, Prášil IT (2007) The role of dehydrins in plant response to cold. Biol Plant 51:601–617CrossRefGoogle Scholar
  33. Lamb C, Dixon RA (1997) The oxidative burst in plant disease resistance. Annu Rev Plant Physiol Plant Mol Biol 48:251–275PubMedCrossRefGoogle Scholar
  34. Lin C, Shalitin D (2003) Cryptochrome structure and signal transduction. Annu Rev Plant Biol 54:469–496PubMedCrossRefGoogle Scholar
  35. Marcellos H (1977) Wheat frost injury-freezing stress and photosynthesis. Aust J Agr Res 28:557–564CrossRefGoogle Scholar
  36. Metwally A, Safronova V, Belimov A, Dietz K (2005) Genotypic variation of the response to cadmium toxicity in Pisum sativum L. J Exp Bot 56:167–178PubMedGoogle Scholar
  37. Mollá S, Villar-Salvador P, García-Fayos P, Rubira JLP (2006) Physiological and transplanting performance of Quercus ilex L. (holm oak) seedlings grown in nurseries with different winter conditions. Forest Ecol Manag 237:218–226CrossRefGoogle Scholar
  38. Monroy AF, Dryanova A, Malette B, Oren DH, Farajalla MR, Liu WC, Danyluk J, Ubayasena LWC, Kane K, Scoles GJ, Sarhan F, Gulick PJ (2007) Regulatory gene candidates and gene expression analysis of cold acclimation in winter and spring wheat. Plant Mol Biol 64:409–423PubMedCrossRefGoogle Scholar
  39. Nakano T, Suzuki K, Fujimura T, Shinshi H (2006) Genome-wide analysis of the ERF gene family in Arabidopsis and rice. Plant Physiol 140:411–432PubMedCrossRefGoogle Scholar
  40. Nakashima K, Yamaguchi-Shinozaki K (2006) Regulons involved in osmotic stress-responsive and cold stress-responsive gene expression in plants. Physiol Plantarum 126:62–71CrossRefGoogle Scholar
  41. Negishi T, Nakanishi H, Yazaki J, Kishimoto N, Fujii F, Shimbo K, Yamamoto K, Sakata K, Sasaki T, Kikuchi S, Mori S, Nishizawa NK (2002) cDNA microarray analysis of gene expression during Fe-deficiency stress in barley suggests that polar transport of vesicles is implicated in phytosiderophore secretion in Fe-deficient barley roots. Plant J 30:83–94PubMedCrossRefGoogle Scholar
  42. Peng YH, Arora R, Li GW, Wang X, Fessehaie A (2008) Rhododendron catawbiense plasma membrane intrinsic proteins are aquaporins, and their over-expression compromises constitutive freezing tolerance and cold acclimation ability of transgenic Arabidopsis plants. Plant Cell Environ 31:1275–1289PubMedCrossRefGoogle Scholar
  43. Phillips JR, Dunn MA, Hughes MA (1997) mRNA stability and localization of the low-temperature-responsive barley gene family blt14. Plant Mol Biol 33:1013–1023PubMedCrossRefGoogle Scholar
  44. Reinheimer JL, Barr AR, Eglinton JK (2004) QTL mapping of chromosomal regions conferring reproductive frost tolerance in barley (Hordeum vulgare L.). Theor Appl Genet 109:1267–1274PubMedCrossRefGoogle Scholar
  45. Renaut J, Hausman J, Wisniewski ME (2006) Proteomics and low-temperature studies: bridging the gap between gene expression and metabolism. Physiol Plantarum 126:97–109CrossRefGoogle Scholar
  46. Richards KD, Schott EJ, Sharma YK, Davis KR, Richard C, Gardner RC (1998) Aluminum induces oxidative stress genes in Arabidopsis thaliana. Plant Physiol 116:409–418PubMedCrossRefGoogle Scholar
  47. Robertson AJ, Reaney MJT, Wilen RW, Lamb N, Abrams SR, Custa LV (1994) Effects of abscisic acid metabolites and analogs on freezing tolerance and gene expression in bromegrass (Bromus inermis Leyss) cell cultures. Plant Physiol 105:823–830PubMedCrossRefGoogle Scholar
  48. Sakr S, Alves G, Morillon R, Morillon R, Maurel K, Decourteix M, Guilliot A, Fleurat-Lessard P, Julien JL, Chrispeels MJ (2003) Plasma membrane aquaporins are involved in winter embolism recovery in walnut tree. Plant Physiol 133:630–641PubMedCrossRefGoogle Scholar
  49. Sasaki T, Ezaki B, Matsumoto H (2002) A gene encoding multidrug resistance (MDR)-like protein is induced by aluminum and inhibitors of calcium flux in wheat. Plant Cell Physiol 43:177–185PubMedCrossRefGoogle Scholar
  50. Satoh R, Fujita Y, Nakashima K, Shinozaki K, Yamaguchi-Shinozaki K (2004) A novel subgroup of bZIP proteins functions as transcriptional activators in hypoosmolarity-responsive expression of the ProDH gene in Arabidopsis. Plant Cell Physiol 45:309–317PubMedCrossRefGoogle Scholar
  51. Shroyer JP, Mikesell ME, Paulsen GM (1995) Spring freeze injury to Kanasas wheat. Publication C-646. Kansas State University, Manhattan, KSGoogle Scholar
  52. Si J, Wang J, Zhang L, Zhang H, Liu Y, An L (2009) CbCOR15, a cold-regulated gene from alpine Chorispora bungeana, confers cold tolerance in transgenic tobacco. J Plant Biol 52:593–601CrossRefGoogle Scholar
  53. Singh HP, Batish DR, Kohli RK, Arora K (2007) Arsenic-induced root growth inhibition in mung bean (Phaseolus aureus Roxb.) is due to oxidative stress resulting from enhanced lipid peroxidation. Plant Growth Regul 53:65–73CrossRefGoogle Scholar
  54. Snowden KC, Gardner RC (1993) Five genes induced by aluminum in wheat (Triticum aestivum L.) roots. Plant Physiol 103:855–861PubMedCrossRefGoogle Scholar
  55. Stockand JD, Zeltwanger S, Bao HF, Becchetti A, Worrell RT, Eaton DC (2001) S-adenosyl-l-homocysteine hydrolase is necessary for aldosterone-induced activity of epithelial Na+ channels. Am J Physiol Cell Ph 281:C773–C785Google Scholar
  56. Tremblay K, Ouellet F, Fournier J, Danyluk J, Sarhan F (2005) Molecular characterization and origin of novel bipartite cold-regulated ice recrystallization inhibition proteins from cereals. Plant Cell Physiol 46:884–891PubMedCrossRefGoogle Scholar
  57. Tsvetanov S, Ohno R, Tsuda K, Takumi S, Mori N, Atanassov A, Nakamura C (2000) A cold-responsive wheat (Triticum aestivum L.) gene wcor14 identified in a winter-hardy cultivar ‘Mironovska 808’. Genes Genet Syst 75:49–57PubMedCrossRefGoogle Scholar
  58. Uemura M, Cilmour SJ, Thomashow MF, Steponkus PL (1996) Effects of COR6.6 and CORl5am polypeptides encoded by COR (cold-regulated) genes of Arabidopsis thaliana on the freeze-induced fusion and leakage of liposomes. Plant Physiol 111:313–327PubMedCrossRefGoogle Scholar
  59. Van Buskirk HA, Thomashow MF (2006) Arabidopsis transcription factors regulating cold acclimation. Physiol Plantarum 126:72–80CrossRefGoogle Scholar
  60. Winfield MO, Lu C, Wilson ID, Coghill JA, Edwards KJ (2009) Cold- and light-induced changes in the transcriptome of wheat leading to phase transition from vegetative to reproductive growth. BMC Plant Biol 9:55PubMedCrossRefGoogle Scholar
  61. Winfield MO, Lu C, Wilson ID, Coghill JA, Edwards KJ (2010) Plant responses to cold: transcriptome analysis of wheat. Plant Biotechnol J 8:749–771PubMedCrossRefGoogle Scholar
  62. Yang KS, Kim HS, Jin UH, Lee SS, Park J, Lim YP, Pai H (2007) Silencing of NbBTF3 results in developmental defects and disturbed gene expression in chloroplasts and mitochondria of higher plants. Planta 225:1459–1469PubMedCrossRefGoogle Scholar
  63. Yeh S, Griffith M, Xiong F, Yang DSC, Wiseman SB, Sarhan F, Danyluk J, Xue YQ, Hew CL, Doherty-Kirby A, Lajoie G (2000) Chitinase genes responsive to cold encode antifreeze proteins in winter cereals. Plant Physiol 124:1251–1264PubMedCrossRefGoogle Scholar
  64. Yoo SY, Kim Y, Kim SY, Lee JS, Ahn JH (2007) Control of flowering time and cold response by a NAC-Domain protein in Arabidopsis. PLoS ONE 2:e642PubMedCrossRefGoogle Scholar
  65. Zhang ZJ, Huang RF (2010) Enhanced tolerance to freezing in tobacco and tomato overexpression transcription factor TERF2/LeERF2 is modulated by ethylene biosynthesis. Plant Mol Biol 73:241–249PubMedCrossRefGoogle Scholar
  66. Zhong X, Mei X, Li Y, Yoshida H, Zhao P, Wang X, Han L, Hu X, Huang S, Huang J, Sun Z (2008) Changes in frost resistance of wheat young ears with development during jointing stage. J Agron Crop Sci 194:343–349CrossRefGoogle Scholar
  67. Zhou MQ, Shen C, Wu LH, Tang KX, Lin J (2011) CBF-dependent signaling pathway: a key responder to low temperature stress in plants. Crit Rev Biotechnol 31:186–192PubMedCrossRefGoogle Scholar
  68. Zhu JH, Dong CH, Zhu JK (2007) Interplay between cold-responsive gene regulation, metabolism and RNA processing during plant cold acclimation. Curr Opin Plant Biol 10:290–295PubMedCrossRefGoogle Scholar

Copyright information

© Franciszek Górski Institute of Plant Physiology, Polish Academy of Sciences, Kraków 2012

Authors and Affiliations

  • Guozhang Kang
    • 1
  • Gezi Li
    • 1
  • Wenping Yang
    • 2
  • Qiaoxia Han
    • 1
  • Hongzhen Ma
    • 1
  • Yonghua Wang
    • 1
  • Jiangping Ren
    • 1
  • Yunji Zhu
    • 1
  • Tiancai Guo
    • 1
  1. 1.National Engineering Research Centre for Wheat, Key Laboratory of Physiology, Ecology and Genetic Improvement of Food Crops in Henan ProvinceHenan Agricultural UniversityZhengzhouChina
  2. 2.Department of AgricultureHenan Institute of Science and TechnologyXinxiangChina

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