Factors of wheat frost hardiness – ice recrystallization inhibitor proteins

For winter wheat, winter hardiness is one of the complex traits that determine the successful cultivation of this crop, and the responsible genes are recognized as highly significant for breeding work. The accumulation of proteins that prevent ice recrystallization (ice recrystallization inhibition proteins, IRIP) correlates with the survival of winter wheat, which indicates the importance of taking this trait into account when obtaining more frost-resistant varieties. The importance of IRIPs is determined by their ability to integrate into growing ice crystals, which limits the formation of large ice conglomerates in the tissues of winter plants. Wheat IRIPs, which accumulate mainly in the apoplast of leaves and in the crowns during cold acclimation, are characterized by a typical duality of structural organization that determines both the manifestation of IRI activity and anti-pathogenic properties. The wheat IRIP molecule contains at the C-terminus a conserved NxVx(x)G fragment that repeats several times, forming a β-helix responsible for binding to the ice surface; at the N-terminus, there is an LRR sequence typical of pathogen-activated kinases, as well as a guiding signal peptide. The wheat genome contains up to eleven IRI genes. The TaIRI gene promoter contains typical basic cis-activating elements and some elements that respond to abiotic stress and hormones. Isoforms of proteins responsible for protecting against pathogens (pathogenesis related proteins, PRP), which accumulate in winter wheat during cold acclimation, also have IRI activity. The expression of the IRIP and PRP genes positively correlates with the cold resistance of winter wheat plants. According to modern data, the regulation of the IRIP genes and cold-activated PRP genes is ABA-independent, but depends on the presence of jasmonic acid and on some proteomic transcription factors. The review provides examples of the practical use of isolated winter wheat IRIPs. The issue of the factors regulating the activity of the IRIP genes and cold-activated PRPs is the least developed to date. The association of these proteins with the winter hardiness of wheat indicates the prospects for their further study.

1. Ambroise V., Legay S., Guerriero G., Hausman J.F., Cuypers A., Sergeant K. The roots of plant frost hardiness and tolerance. Plant Cell Physiol. 2020;61(1):3-20. doi 10.1093/pcp/pcz196

2. Antikainen M., Griffith M. Antifreeze protein accumulation in freezingtolerant cereals. Physiol Plant. 1997;99(3):423-432. doi 10.1111/ j.1399-3054.1997.tb00556.x

3. Badawi M., Reddy Y.V., Agharbaoui Z., Tominaga Y., Danyluk J., Sarhan F., Houde M. Structure and functional analysis of wheat ICE (inducer of CBF expression) genes. Plant Cell Physiol. 2008;49(8): 1237-1249. doi 10.1093/pcp/pcn100

4. Båga M., Chodaparambil S.V., Limin A.E., Pecar M., Fowler D.B., Chibbar R.N. Identification of quantitative trait loci and associated candidate genes for low-temperature tolerance in cold-hardy winter wheat. Funct Integr Genomics. 2007;7(1):53-68. doi 10.1007/s10142-006-0030-7

5. Bayer-Giraldi M., Weikusat I., Besir H., Dieckmann G. Characterization of an antifreeze protein from the polar diatom Fragilariopsis cylindrus and its relevance in sea ice. Cryobiology. 2011;63(3):210219. doi 10.1016/j.cryobiol.2011.08.006

6. Boonsupthip W., Lee T.-C. Application of antifreeze protein for food preservation: effect of type III antifreeze protein for preservation of gel-forming of frozen and chilled actomyosin. J Food Sci. 2003; 68(5):1804-1809. doi 10.1111/j.1365-2621.2003.tb12333.x

7. Cao Y., Hu G., Zhuang M., Yin J., Wang X. Molecular cloning and functional characterization of TaIRI9 gene in wheat (Triticum aestivum L.). Gene. 2021;791:145694. doi 10.1016/j.gene.2021.145694

8. Case A.J., Skinner D.Z., Garland-Campbell K.A., Carter A.H. Freezing tolerance-associated quantitative trait loci in the brundage × coda wheat recombinant inbred line population. Crop Sci. 2014;54(3): 982-992. doi 10.2135/cropsci2013.08.0526

9. Chinnusamy V., Ohta M., Kanrar S., Lee B.H., Hong X., Agarwal M., Zhu J.K. ICE1: a regulator of cold-induced transcriptome and freezing tolerance in Arabidopsis. Genes Dev. 2003;17(8):1043-1054. doi 10.1101/gad.1077503

10. Chow-Shi-Yée M., Briard J.G., Grondin M., Averill-Bates D.A., Ben R.N., Ouellet F. Inhibition of ice recrystallization and cryoprotective activity of wheat proteins in liver and pancreatic cells. Protein Sci. 2016;25(5):974-986. doi 10.1002/pro.2903

11. Chow-Shi-Yée M., Grondin M., Ouellet F., Averill-Bates D.A. Control of stress-induced apoptosis by freezing tolerance-associated wheat proteins during cryopreservation of rat hepatocytes. Cell Stress Chaperones. 2020;25(6):869-886. doi 10.1007/s12192-020-01115-y

12. Chun J., Yu X., Griffith M. Genetic studies of antifreeze proteins and their correlation with winter survival in wheat. Euphytica. 1998;102: 219-226. doi 10.1023/A:1018333730936

13. DeVries A.L., Wohlschlag D.E. Freezing resistance in some Antarctic fishes. Science. 1969;163(3871):1073-1075. doi 10.1126/science.163.3871.1073

14. Duman J.G., Olsen T.M. Thermal hysteresis protein activity in bacteria, fungi, and phylogenetically diverse plants. Cryobiology. 1993; 30(3):322-328. doi 10.1006/cryo.1993.1031

15. Duman J.G., Wisniewski M.J. The use of antifreeze proteins for frost protection in sensitive crop plants. Env Exp Bot. 2014;106:60-69. doi 10.1016/j.envexpbot.2014.01.001

16. Ewart K.V., Lin Q., Hew C.L. Structure, function and evolution of antifreeze proteins. Cell Mol Life Sci. 1999;55(2):271-283. doi 10.1007/s000180050289

17. Gaudet D.A., Laroche A. Winter survival of cereals parasitized by Snow Mold. In: Li P.H., Chen T.H.H. (Eds) Plant Cold Hardiness. Boston: Springer, 1997;331-342. doi 10.1007/978-1-4899-0277-1_31

18. Gaudet D.A., Laroche A., Frick M., Davoren J., Puchalski B., Ergon Å. Expression of plant defence-related (PR-protein) transcripts during hardening and dehardening of winter wheat. Physiol Mol Plant Pathol. 2000;57(1):15-24. doi 10.1006/pmpp.2000.0275

19. Griffith M., Yaish M.W. Antifreeze proteins in overwintering plants: a tale of two activities. Trends Plant Sci. 2004;9(8):399-405. doi 10.1016/j.tplants.2004.06.007

20. Griffith M., Ala P., Yang D.S., Hon W.C., Moffatt B.A. Antifreeze protein produced endogenously in winter rye leaves. Plant Physiol. 1992;100(2):593-596. doi 10.1104/pp.100.2.593

21. Griffith M., Lumb C., Wiseman S.B., Wisniewski M., Johnson R.W., Marangoni A.G. Antifreeze proteins modify the freezing process in planta. Plant Physiol. 2005;138(1):330-340. doi 10.1104/pp.104.058628

22. Grondin M., Hamel F., Averill-Bates D.A., Sarhan F. Wheat proteins improve cryopreservation of rat hepatocytes. Biotechnol Bioeng. 2009;103(3):582-591. doi 10.1002/bit.22270

23. Hannah M.A., Heyer A.G., Hincha D.K. A global survey of gene regulation during cold acclimation in Arabidopsis thaliana. PLoS Genet. 2005;1(2):e26. doi 10.1371/journal.pgen.0010026

24. He K., Wu Y. Receptor-like kinases and regulation of plant innate immunity. Enzymes. 2016;40:105-142. doi 10.1016/bs.enz.2016.09.003

25. Herman E.M., Rotter K., Premakumar R., Elwinger G., Bae H., EhlerKing L., Chen S., Livingston D.P. 3rd. Additional freeze hardiness in wheat acquired by exposure to –3 degrees C is associated with extensive physiological, morphological, and molecular changes. J Exp Bot. 2006;57(14):3601-3618. doi 10.1093/jxb/erl111

26. Hon W.C., Griffith M., Mlynarz A., Kwok Y.C., Yang DSC. Antifreeze proteins in winter rye are similar to pathogenesis-related proteins. Plant Physiol. 1995;109(3):879-889. doi 10.1104/pp.109.3.879

27. Houde M., Belcaid M., Ouellet F., Danyluk J., Monroy A.F., Dryanova A., Gulick P., Bergeron A., Laroche A., Links M.G., MacCarthy L., Crosby W.L., Sarhan F. Wheat EST resources for functional genomics of abiotic stress. BMC Genomics. 2006;7:149. doi 10.1186/1471-2164-7-149

28. Huang T., Duman J.G. Cloning and characterization of a thermal hysteresis (antifreeze) protein with DNA-binding activity from winter bittersweet nightshade, Solanum dulcamara. Plant Mol Biol. 2002; 48(4):339-350. doi 10.1023/A:1014062714786

29. Jain D., Khurana J.P. Role of pathogenesis-related (PR) proteins in plant defense mechanism. In: Singh A., Singh I. (Eds) Molecular Aspects of Plant-Pathogen Interaction. Singapore: Springer, 2018;265-281. doi 10.1007/978-981-10-7371-7_12

30. Janech M.G., Krell A., Mock T., Raymond J.A. Ice-binding proteins from sea ice diatoms (Bacillariophyceae). J Phycol. 2006;42(2):410416. doi 10.1111/j.1529-8817.2006.00208.x

31. Jarząbek M., Pukacki P.M., Nuc K. Cold-regulated proteins with potent antifreeze and cryoprotective activities in spruces (Picea sp.). Cryobiology. 2009;58(3):268-274. doi 10.1016/j.cryobiol.2009.01.007

32. Jin Y.N., Bai L.P., Guan S.X., Zhong M., Ma H., Wang S., Guo Z.F. Identifcation of an ice recrystallisation inhibition gene family in winter-hardy wheat and its evolutionary relationship to other members of the Triticeae. J Agron Crop Sci. 2018;204(4):400-413. doi 10.1111/jac.12270

33. Jin Y., Ding X., Li J., Guo Z. Isolation and characterization of wheat ice recrystallisation inhibition gene promoter involved in low temperature and methyl jasmonate responses. Physiol Mol Biol Plants. 2022;28(11-12):1969-1979. doi 10.1007/s12298-022-01257-6

34. Juurakko C.L., diCenzo G.C., Walker V.K. Brachypodium antifreeze protein gene products inhibit ice recrystallisation, attenuate ice nucleation, and reduce immune response. Plants. 2022;11(11):1475. doi 10.3390/plants11111475

35. Kang G., Li G., Yang W., Han Q., Ma H., Wang Y., Ren J., Zhu Y., Guo T. Transcriptional profile of the spring freeze response in the leaves of bread wheat (Triticum aestivum L.). Acta Physiol Plant. 2013;35(02):575-587. doi 10.1007/s11738-012-1099-3

36. Knight C.A., Wen D., Laursen R.A. Nonequilibrium antifreeze peptides and the recrystallization of ice. Cryobiology. 1995;32(1):23-34. doi 10.1006/cryo.1995.1002

37. Kontogiorgos V., Regand A., Yada R., Goff H.D. Isolation and characterization of ice structuring proteins from cold-acclimated winter wheat grass extract for recrystallization inhibition in frozen foods. J Food Biochem. 2007;31(2):139-160. doi 10.1111/j.1745-4514.2007.00112.x

38. Kostyaev A.A., Martusevich A.K., Andreev A.A. Toxicity of cryoprotectants and cryoconservants on their basis for blood components and bone marrow (review article). Nauchnoe Obozrenie. Meditsinskie Nauki = Scientific Review. Medical Sciences. 2016;6:54-74 (in Russian)

39. Kristiansen E., Pedersen S.A., Ramløv H., Zachariassen K.E. Antifreeze activity in the cerambycid beetle Rhagium inquisitor. J Comp Physiol B. 1999;169:55-60. doi 10.1007/s003600050193

40. Kristiansen E., Ramløv H., Højrup P., Pedersen S.A., Hagen L., Zachariassen K.E. Structural characteristics of a novel antifreeze protein from the longhorn beetle Rhagium inquisitor. Insect Biochem Mol Biol. 2011;41(2):109-117. doi 10.1016/j.ibmb.2010.11.002

41. Kruse E.B., Carle S.W., Wen N., Skinner D.Z., Murray T.D., GarlandCampbell K.A., Carter A.H. Genomic regions associated with tolerance to freezing stress and snow mold in winter wheat. G3 (Bethesda). 2017;7(3):775-780. doi 10.1534/g3.116.037622

42. Kurepin L.V., Dahal K.P., Savitch L.V., Singh J., Bode R., Ivanov A.G., Hurry V., Hüner N.P. Role of CBFs as integrators of chloroplast redox, phytochrome and plant hormone signaling during cold acclimation. Int J Mol Sci. 2013;14(6):12729-12763. doi 10.3390/ijms140612729

43. Liu X., Pan Y., Liu F., He Y., Zhu Q., Liu Z., Zhan X., Tan S. A review of the material characteristics, antifreeze mechanisms, and applications of cryoprotectants (CPAs). J Nanomater. 2021;2021(1):9990709. doi 10.1155/2021/9990709

44. Livingston D.P. 3rd, Bertrand A., Wisniewski M., Tisdale R., Tuong T., Gusta L.V., Artlip T. Factors contributing to ice nucleation and sequential freezing of leaves in wheat. Planta. 2021;253(6):124. doi 10.1007/s00425-021-03637-w

45. Macháčcková I., Hanisova A., Krekule J. Levels of ethylene, ACC, MACC, ABA and proline as indicators of cold hardening and frost resistance in winter wheat. Physiol Plant. 1989;76(4):603-607. doi 10.1111/j.1399-3054.1989.tb05486.x

46. McHale L., Tan X., Koehl P., Michelmore R.W. Plant NBS-LRR proteins: adaptable guards. Genome Biol. 2006;7(4):212. doi 10.1186/gb-2006-7-4-212

47. Middleton A.J., Brown A.M., Davies P.L., Walker V.K. Identification of the ice-binding face of a plant antifreeze protein. FEBS Lett. 2009; 583(4):815-819. doi 10.1016/j.febslet.2009.01.035

48. Middleton A.J., Marshal C.B., Faucher F., Bar-Dolev M., Braslavsky I., Campbell R.L., Walker V.K., Davies P.L. Antifreeze protein from freeze-tolerant grass has a beta-roll fold with an irregularly structured ice-binding site. J Mol Biol. 2012;416(5):713-724. doi 10.1016/j.jmb.2012.01.032

49. Monroy A.F., Dryanova A., Malette B., Oren D.H., Ridha Farajalla M., Liu W., Danyluk J., Ubayasena L.W., Kane K., Scoles G.J., Sarhan F., Gulick P.J. Regulatory gene candidates and gene expression analysis of cold acclimation in winter and spring wheat. Plant Mol Biol. 2007;64(4):409-423. doi 10.1007/s11103-007-9161-z

50. Pandey S.P., Somssich I.E. The role of WRKY transcription factors in plant immunity. Plant Physiol. 2009;150(4):1648-1655. doi 10.1104/pp.109.138990

51. Puchkov E.O. Biogenic control of ice formation. Priroda. 2017;2: 27-37 (in Russian)

52. Regand A., Goff H.D. Ice recrystallization inhibition in ice cream as affected by ice structuring proteins from winter wheat grass. J Dairy Sci. 2006;89(1):49-57. doi 10.3168/jds.S0022-0302(06)72068-9

53. Sandve S.R., Rudi H., Asp T., Rognli O.A. Tracking the evolution of a cold stress associated gene family in cold tolerant grasses. BMC Evol Biol. 2008;8:245. doi 10.1186/1471-2148-8-245

54. Sauter M. Phytosulfokine peptide signaling. J Exp Bot. 2015;66(17): 5161-5169. doi 10.1093/jxb/erv071

55. Sutka J. Genetic control of frost tolerance in wheat (Triticum aestivum L.). Euphytica. 1994;77:277-282. doi 10.1007/BF02262642

56. Sutka J. Genes for frost resistance in wheat. Euphytica. 2001;119:169177. doi 10.1023/A:1017520720183

57. Sutka J., Galiba G., Snape J.W. Inheritance of frost resistance in wheat (Triticum aestivum L.). Acta Agron Hung. 1997;45:257-263

58. Talanova V.V., Titov A.F., Topchieva L.V., Malysheva I.E., VenzhikY.V., Frolova S.A. Expression of genes encoding the WRKY transcription factor and heat shock proteins in wheat plants during cold hardening. Dokl Biol Sci. 2008;423:440-442. doi 10.1134/s0012496608060215

59. Tchagang A.B., Fauteux F., Tulpan D., Pan Y. Bioinformatics identification of new targets for improving low temperature stress tolerance in spring and winter wheat. BMC Bioinformatics. 2017;18(1):174. doi 10.1186/s12859-017-1596-x

60. Tremblay K., Ouellet F., Fournier J., Danyluk J., Sarhan F. Molecular characterization and origin of novel bipartite cold-regulated ice recrystallization inhibition proteins from cereals. Plant Cell Physiol. 2005;46(6):884-891. doi 10.1093/pcp/pci093

61. Urrutia M.E., Duman J.G., Knight C.A. Plant thermal hysteresis proteins. Biochem Biophys Acta. 1992;1121(1-2):199-206. doi 10.1016/0167-4838(92)90355-h

62. Vaitkevičiūte G., Aleliūnas A., Brazauskas G., Armonienè R. Deacclimation and reacclimation processes in winter wheat: novel perspectives from time-series transcriptome analysis. Front Plant Sci. 2024; 15:1395830. doi 10.3389/fpls.2024.1395830

63. Vanková R., Kosová K., Dobrev P., Vítámvás P., Trávníčková A., Cvikrová M., Pešek B., Gaudinová A., Přerostová S., Musilová J., Galiba G., Prasil I.T. Dynamics of cold acclimation and complex phytohormone responses in Triticum monococcum lines G3116 and DV92 differing in vernalization and frost tolerance level. Env Exp Bot. 2014;101:12-25. doi 10.1016/j.envexpbot.2014.01.002

64. Voets I.K. From ice-binding proteins to bio-inspired antifreeze materials. Soft Matter. 2017;13(28):4808-4823. doi 10.1039/c6sm02867e

65. Wang W., Hao Q., Wang W., Li Q., Wang W. The genetic characteristics in cytology and plant physiology of two wheat (Triticum aestivum) near isogenic lines with different freezing tolerances. Plant Cell Rep. 2017;36(11):1801-1814. doi 10.1007/s00299-017-2195-z

66. Wang W., Wang X., Zhang J., Huang M., Cai J., Zhou Q., Dai T., Jiang D. Salicylic acid and cold priming induce late-spring freezing tolerance by maintaining cellular redox homeostasis and protecting photosynthetic apparatus in wheat. Plant Growth Regul. 2020;90:109-121. doi 10.1007/s10725-019-00553-8

67. Wang W., Wang X., Huang M., Cai J., Zhou Q., Dai T., Jiang D. Alleviation of field low-temperature stress in winter wheat by exogenous application of salicylic acid. J Plant Growth Regul. 2021;40:811-823. doi 10.1007/s00344-020-10144-x

68. Wang X., Geng H., Wu D., Wang L., Zhang N., Wang W., Yu D. Isolation of ice structuring proteins from winter wheat in frigid region (Dongnongdongmai1) and the effect on freeze-thaw stability of dough. Food Res Int. 2024;197(Pt. 2):115295. doi 10.1016/j.foodres.2024.115295

69. Willick I.R., Takahashi D., Fowler D.B., Uemura M., Tanino K.K. Tissue-specific changes in apoplastic proteins and cell wall structure during cold acclimation of winter wheat crowns. J Exp Bot. 2018; 69(5):1221-1234. doi 10.1093/jxb/erx450

70. Willick I.R., Gusta L.V., Fowler D.B., Tanino K.K. Ice segregation in the crown of winter cereals: evidence for extraorgan and extratissue freezing. Plant Cell Environ. 2019;42(2):701-716. doi 10.1111/pce.13454

71. Winfield M.O., Lu C., Wilson I.D., Coghill J.A., Edwards K.J. Plant responses to cold: transcriptome analysis of wheat. Plant Biotechnol J. 2010;8(7):749-771. doi 10.1111/j.1467-7652.2010.00536.x

72. Wisniewski M., Willick I.R., Duman J.G., Livingston D.P. 3rd, Newton S.S. Plant antifreeze proteins. In: Ramløv H., Friis D. (Eds) Antifreeze Proteins. Vol. 1. Cham: Springer, 2020;189-226. doi 10.1007/978-3-030-41929-5_7

73. Xue-Xuan X., Hong-Bo S., Yuan-Yuan M., Gang X., Jun-Na S., DongGang G., Cheng-Jiang R. Biotechnological implications from abscisic acid (ABA) roles in cold stress and leaf senescence as an important signal for improving plant sustainable survival under abiotic-stressed conditions. Crit Rev Biotechnol. 2010;30(3):222-230. doi 10.3109/07388551.2010.487186

74. Yang T., Zhang Y., Guo L., Li D., Liu A., Bilal M., Xie C., Yang R., Gu Z., Jiang D., Wang P. Antifreeze polysaccharides from wheat bran: the structural characterization and antifreeze mechanism. Biomacromolecules. 2024;25(7):3877-3892. doi 10.1021/acs.biomac.3c00958

75. Yu X.M., Griffith M., Wiseman S.B. Ethylene induces antifreeze activity in winter rye leaves. Plant Physiol. 2001;126(3):1232-1240. doi 10.1104/pp.126.3.1232

76. Zheng X., Shi M., Wang J., Yang N., Wang K., Xi J., Wu C., Xi T., Zheng J., Zhang J. Isoform sequencing provides insight into freezing response of common wheat (Triticum aestivum L.). Front Genet. 2020;11:462. doi 10.3389/fgene.2020.00462