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Estimating Ice Encasement Tolerance of Herbage Plants

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Plant Cold Acclimation

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1166))

Abstract

One of the key stresses acting on herbage plants during winter is ice encasement, when plants are enclosed in compact ice and turn from aerobic to anaerobic respiration. The cause of cell death is related to the accumulation of metabolites to toxic levels during winter and perhaps also to production of reactive oxygen species (ROS) when plants escape from long-lasting ice cover. The process of ice encasement damage has been studied by sampling studies, indirect measurements of ice tolerance, field tests and provocation methods by increasing stress in the field artificially, thus increasing the ice stress. Here we describe a laboratory method to measure ice encasement tolerance. This is the most common and effective way to measure ice encasement tolerance of large plant material. Plants are raised from seeds (or taken from the field), cold acclimated, usually at +2 °C under short day conditions, in a greenhouse or growth chamber (or in the field during fall). Plants are submerged in cold water in beakers and frozen encased in ice, usually at −2 °C. Plants are kept enclosed in ice at this temperature. Samples are taken at intervals, depending on species and tolerance of plant material, and put smoothly to regrowth. Damage is then evaluated after a suitable time of regeneration.

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References

  1. Griffith M, Gudleifsson BE, Fukuta N (2001) Abiotic stresses in overwintering crops. In: Iriki N, Gaudet DA, Tronsmo AM et al (eds) Low temperature plant microbe interactions under snow. Hokkaido National Agricultural Experiment Station, Hitsujigaoka, Toytrira, Sapporo, Japan, pp 101–114

    Google Scholar 

  2. Gudleifsson BE, Larsen A (1993) Ice encasement as a component of winter kill in herbage plants. In: Christersson L, Li PH (eds) Advances in plant cold hardiness. CRC Press, Boca Raton, pp 229–249

    Google Scholar 

  3. Gudleifsson BE (2010) Ice tolerance and metabolite accumulation in herbage crops in Iceland and impact of climate change. Icel Agr Sci 23:111–122

    Google Scholar 

  4. Höglind M, Bakken AK, Jörgensen M et al (2010) Tolerance to frost and ice encasement in cultivars of timothy and perennial ryegrass during winter. Grass Forage Sci 65:431–445

    Article  Google Scholar 

  5. Rakitina ZG (1965) The permeability of ice for O2 and CO2 in connection with a study of the reason for winter cereal mortality under the ice crust. Sov Plant Physiol 12:795–803

    Google Scholar 

  6. Rakitina ZG (1970) Effect of an ice crust on gas composition of the internal atmosphere in winter wheat. Sov Plant Physiol 17:755–759

    Google Scholar 

  7. Andrews CJ, Pomeroy MK (1990) Low temperature anaerobiosis in ice encasement damage to winter cereals. In: Jackson MB, Davies DD, Lambers H (eds) Plant life under oxygen deprivation. SPS Academic Publishing, The Hague, The Netherlands, pp 85–99

    Google Scholar 

  8. Castonguay Y, Thibault C, Rochette P et al (2009) Physiological responses of annual bluegrass to contrasted levels of O2 and CO2 at low temperatures. Crop Sci 49:671–689

    Article  Google Scholar 

  9. Andrews CJ (1977) Accumulation of ethanol in ice-encased winter cereals. Crop Sci 17:157–161

    Article  CAS  Google Scholar 

  10. Gudleifsson BE (1997) Estimating ice encasement tolerance in the laboratory. In: Molecular and physiological aspects of cold and chilling tolerance of northern plants. Proceedings of the Finish-Japanese Workshop, Agricultural Research Center of Finland, Jokioinen, pp 14–15

    Google Scholar 

  11. Gudleifsson BE (2009) Ice encasement damage on grass crops and alpine plants in Iceland – impact of climate change. In: Wisniewski M, Tanino K, Gusta L (eds) Plant cold hardiness. From the laboratory to the field. CAB International, Wallingford, UK, pp 163–172

    Chapter  Google Scholar 

  12. Brandsæter LO, Haugland E, Helgheim E et al (2004) Identification of phytotoxic substances in soil following winter injury of grasses as estimated by a bioassay. Can J Plant Sci 85:115–123

    Article  Google Scholar 

  13. Aamlid TS, Landschoot PJ, Huff DR (2009) Tolerance to simulated ice encasement and Microdochium nivale in USA selections of greens-type Poa annua. Acta Agric Scand Sect B Soil Plant Sci 59:170–178

    CAS  Google Scholar 

  14. Hetherington PR, McKersie BD, Borochov A (1987) Ice encasement injury to microsomal membranes from winter wheat crowns. I. Comparison of membrane properties after lethal ice encasement and during a post-thaw period. Plant Physiol 85:1068–1072

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  15. Hetherington PR, Broughton HL, McKersie BD (1988) Ice encasement injury to microsomal membranes from winter wheat crowns. II. Changes in membrane lipids during ice encasement. Plant Physiol 86:740–743

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  16. Andrews CJ, Pomeroy MK (1989) Physiological properties of plants affecting ice encasement tolerance. Icel Agr Sci 2:41–51

    Google Scholar 

  17. Andrews CJ (1996) How do plants survive ice? Ann Bot 78:529–536

    Article  CAS  Google Scholar 

  18. Bertrand A, Gastonguay Y, Nadeau P et al (2001) Molecular and biochemical responses of perennial forage crops to oxygen deprivation at low temperatures. Plant Cell Environ 24:1085–1093

    Article  CAS  Google Scholar 

  19. Tanino KK, McKersie BD (1985) Injury within the crown of winter wheat seedlings after freezing and icing stress. Can J Bot 63:432–436

    Article  Google Scholar 

  20. McKersie BD, Leshem YY (1994) Stress and stress coping in cultivated plants. Kluwer Academic, Dordrecht

    Book  Google Scholar 

  21. Andrews CJ, Pomeroy MK (1975) Survival and cold hardiness of winter wheats during partial and total ice immersion. Crop Sci 15:561–566

    Article  Google Scholar 

  22. Andrews CJ, Pomeroy MK (1981) The effect of flooding pretreatment on cold hardiness and survival of winter cereals in ice encasement. Can J Plant Sci 61:507–513

    Article  Google Scholar 

  23. Gudleifsson BE, Andrews CJ, Bjornsson H (1986) Cold hardiness and ice tolerance of pasture grasses grown and tested in controlled environments. Can J Plant Sci 66:601–608

    Article  Google Scholar 

  24. Bowley SR, McKersie BD (1990) Relationships among freezing, low temperature flooding, and ice encasement tolerance in alfalfa. Can J Plant Sci 70:227–235

    Article  Google Scholar 

  25. Preece C, Callaghan TV, Phoenix GK (2012) Impacts of winter icing events on the growth, phenology and physiology of sub-arctic dwarf shrubs. Physiol Plant 146:460–473

    Article  CAS  PubMed  Google Scholar 

  26. Pulli S, Hjortsholm K, Larsen A et al (1996) Development and evaluation of laboratory testing methods for winter hardiness breeding, vol 32. Nordic Gene Bank, Alnarp, Sweden, p 98

    Google Scholar 

  27. Baadshaug OH (1973) En vurdering av forskjellige metoder for overvintringsundersökelser i eng og beitevekster. [An evaluation of various methods for investigating wintering of meadow and pasture plants]. Forskn fors Landbr 24:221–234, In Norwegian with English Summary

    Google Scholar 

  28. Voisey PW, Moulton F (1968) Precise temperature control for a domestic freezer. Can J Plant Sci 49:107–110

    Article  Google Scholar 

  29. Baadshaug OH (1973) Effects of soil type and soil compaction on the wintering of three grass species under different wintering conditions. Acta Agric Scand 23:77–86

    Article  Google Scholar 

  30. Gudleifsson BE (1971) Overvintringsskadar i grasmark på Island, omfang og årsaker. [Extent and causes of winter-damages in Icelandic grasslands]. Lisensiatoppgave, Norges landbrukshögskole, p 130, [In Norwegian with English Summary]

    Google Scholar 

  31. Andrews CJ, Seaman WL, Pomeroy MK (1984) Changes in cold hardiness, ice tolerance and total carbohydrates of winter wheat under various cutting regimes. Can J Plant Sci 64:547–558

    Article  CAS  Google Scholar 

  32. McKersie BD, Hunt LA (1987) Genotypic differences in tolerance of ice encasement, low temperature flooding, and freezing in winter wheat. Crop Sci 27:860–863

    Article  Google Scholar 

  33. Larsen A (1986) Test methods for wintering characters. NJF seminar Nr.84 Lantbruksväxternas övervintring, pp 149–166

    Google Scholar 

  34. Sakai A, Larcher W (1987) Frost survival of plants. Responses and adaptation to freezing stress. Springer, Berlin, Heidelberg

    Google Scholar 

  35. Larcher W (1985) Schädigung der Pflanzen durch Bodenfrost und Schneebedeckung. In: Larcher W, Häckel H, Sakai A (eds) Handbuch der Pflanzenkrankheiten. I, 5. Die Nichtparasitären Krankheiten. Verlag Paul Parey, Berlin, pp 274–287, (In German)

    Google Scholar 

  36. Fridriksson S (1954) Rannsóknir á kali túna árin 1951 og 1952. [Winter injury of plants in Icelandic hayfields]. Rit Landb.Deild. B-7, 72s, In Icelandic with English Summary

    Google Scholar 

  37. Andersen IL (1963) Overvintringsundersøkelser i eng i Nord-Norge. II. Noen undersøkelser over is og vannskader i eng. [Investigations on the wintering of meadow plants in northern Norway. II. Some investigations on damages caused by ice and water choking on meadows]. Forsk Fors Landbr 14:639–669, In Norwegian with English Summary

    Google Scholar 

  38. Årsvoll K (1973) Winter damage in Norwegian grasslands, 1968–1971. Meld Norges Landbrukshögskole 52(3):21

    Google Scholar 

  39. Hakamata T, Noshiro M, Hirashima T, Nose I (1978) Investigation of actual condition on the winter-killing of pasture species in the Nemuro-Kushiro district. Exploration of factors by the quantification No 1. J Jpn Grassl Sci 23:280–288

    Google Scholar 

  40. Levitt J (1980) Responses of plants to environmental stresses. I. Chilling, freezing, and high temperature stresses. Academic Press, New York

    Google Scholar 

  41. Gusta LV, Fowler DB, Tyler MJ (1983) An evaluation of several chemical tests as possible selection measures for winter hardiness in wheat. Can J Plant Sci 63:115–119

    Article  CAS  Google Scholar 

  42. Fowler DB, Gusta LV, Tyler NJ (1981) Selection for winter hardiness in wheat. III. Screening methods. Crop Sci 21:896–901

    Article  CAS  Google Scholar 

  43. Andersen IL (1960) Overvintringsundersökelser i eng i Nord-Norge. I. [Investigation on the wintering of meadow plants in Northern-Norway]. Forskn fors Landbr 11:635–660, In Norwegian with English Summary

    Google Scholar 

  44. Brink RA, Keller W, Eisenhart C (1939) Differential survival of alfalfa under an ice sheet. J Agr Res 59:59–71

    Google Scholar 

  45. Gudleifsson BE, Sigvaldason J (1972) Um kal og kalskemmdir. II. Kalskemmdir í tilraunareitum Tilraunastöðvarinnar á Akureyri. [On winter damage. II. Winter damage on experimental plots in Akureyri]. Ársrit Ræktunarf Norðurlands 69:84–101, In Icelandic with English Summary

    Google Scholar 

  46. Fowler DB (1979) Selection for winter hardiness in wheat. II. Variation in field trials. Crop Sci 19:773–775

    Article  Google Scholar 

  47. Beard JB (1965) Effects of ice covers in the field on two perennial grasses. Crop Sci 5:139–140

    Article  Google Scholar 

  48. Sjøseth H (1959) Studies on ice encasement in strains of red clover (Trifolium pratense) and timothy (Phleum pratense). Acta Agric Scand 9:292–298

    Article  Google Scholar 

  49. Sjøseth H (1971) Vinterhardførhet hos ulike eng- og beitevekster. [Winter hardiness in different meadow and pasture plants]. Meld Norges Landbrukshögskole 50:39, In Norwegian with English Summary

    Google Scholar 

  50. Andrews CJ, Pomeroy MK (1977) Changes in survival of winter cereals due to ice cover and other simulated winter conditions. Can J Plant Sci 57:1141–1149

    Article  Google Scholar 

  51. Andrews CJ, Pomeroy MK, Seaman WL (1986) The response of fall-sown cereals to winter stress in eastern Ontario. Can J Plant Sci 66:25–37

    Article  Google Scholar 

  52. Gudleifsson BE (1997) Survival and metabolite accumulation by seedlings and mature plants of timothy grass during ice encasement. Ann Bot 79(Suppl A):93–96

    Article  CAS  Google Scholar 

  53. Sprague MA, Graber LF (1943) Ice sheet injury to alfalfa. J Am Soc Agron 35:881–894

    Article  CAS  Google Scholar 

  54. Smith D (1952) The survival of winter-hardened legumes encased in ice. Agron J 44:469–473

    Article  Google Scholar 

  55. Beard JB (1964) Effects of ice, snow and water covers on Kentucky bluegrass, annual bluegrass and creeping bentgrass. Crop Sci 4:638–640

    Article  Google Scholar 

  56. Freyman S, Brink VC (1967) Nature of ice-sheet injury to alfalfa. Agron J 59:557–560

    Article  Google Scholar 

  57. Andrews CJ, Pomeroy MK, de la Roche IA (1974) Changes in cold hardiness of overwintering winter wheat. Can J Plant Sci 54:9–15

    Article  Google Scholar 

  58. Andrews CJ, Gudleifsson BE (1983) A comparison of cold hardiness and ice encasement tolerance of timothy grass and winter wheat. Can J Plant Sci 63:429–435

    Article  Google Scholar 

  59. Gudleifsson BE (1986) Måling av isdekketoleranse hos gras i laboratoriet. [Evaluating ice tolerance of plants in the laboratory]. NJF seminar Nr. 84 Lantbruksväxternas övervintring. pp 171–175, [In Norwegian]

    Google Scholar 

  60. Bolduc R (1976) Tecnique pour echantillonner les racines de plantes dans le sol gelé et enneige. [Method to collect samples of plants under snow and ice]. Can J Plant Sci 56:633–638, In French with English Summary

    Article  Google Scholar 

  61. Pomeroy MK, Fowler DB (1973) Use of lethal dose temperature estimates as indices of frost tolerance for wheat cold acclimated under natural and controlled environments. Can J Plant Sci 53:489–494

    Article  Google Scholar 

  62. Brule-Babel AL, Fowler DB (1989) Use of controlled environments for winter cereal cold hardiness evaluation: controlled freeze tests and tissue water content as prediction tests. Can J Plant Sci 69:355–366

    Article  Google Scholar 

  63. Smillie RM, Hetherington SE (1983) Stress tolerance and stress-induced injury in crop plants measured by chlorophyll fluorescence in vivo. Chilling, freezing, ice cover, heat and high light. Plant Physiol 72:1043–1050

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  64. Samygin GA, Rakitina ZG, Livshin AZ (1971) Resistance of winter wheat tissues to freezing and desiccation under adverse gas regime. Dokl Bot Sci 196–198:77–80

    Google Scholar 

  65. Samygin GA, Rakitina ZG, Livshin AZ (1973) Influence of an adverse gaseous regime and ice-cold bark on the water-retaining capacity of the leaves and tillering nodes of hardened winter wheat. Dokl Bot Sci 209:60–62

    Google Scholar 

  66. Hope HJ, Comeau A, Hasty P (1984) Ice encasement tolerance of prairieland wild ryegrass, orbit tall wheatgrass and puma rye grown under controlled environments. Cereal Res Com 12:101–103

    Google Scholar 

  67. Larsen A (1978) Freezing tolerance in grasses. Methods for testing in controlled environments. Meldinger fra Norges Landbrukshögskole 57(23):56

    Google Scholar 

  68. Gudleifsson BE, Björnsson H (1989) Methods for estimating ice-encasement tolerance of grasses in the laboratory. Icel Agr Sci 2:99–103

    Google Scholar 

  69. Tronsmo AM, Svendsen S (1989) Ice encasement in timothy and cocksfoot – a possible screening method for application in breeding programs. Icel Agr Sci 2:105–107

    Google Scholar 

  70. McKersie BD, McDermott BM, Hunt LA et al (1982) Changes in carbohydrate levels during ice encasement and flooding of winter cereals. Can J Bot 60:1822–1826

    Article  CAS  Google Scholar 

  71. Poysa VW (1984) The genetic control of low temperature, ice-encasement, and flooding tolerances by chromosomes 5A, 5B and 5D in wheat. Cereal Res Comm 12:135–141

    Google Scholar 

  72. McKersie BD (1983) Types of winter stresses – do winter wheat cultivars respond differently? In: Fowler DB, Gusta LV, Slinkard AE et al (eds) New frontiers in winter wheat production, Western Canada winter wheat conference., pp 26–38

    Google Scholar 

  73. Sjøseth H (1957) Undersøkelser over frostherdighet hos engvekster. [Studies on frost hardiness in meadow plants]. Forskn fors Landbr 8:77–98, In Norwegian with English Summary

    Google Scholar 

  74. Freyman S (1969) Role of stubble in the survival of certain ice-covered forages. Agron J 61:105–107

    Article  Google Scholar 

  75. Tompkins DK, Ross JB, Moroz DL (2004) Effects of ice cover on annual bluegrass and creeping bentgrass putting greens. Crop Sci 44:2157–2179

    Article  Google Scholar 

  76. Pukacki PM, McKersie BD (1990) Supercooling and ice nucleation events in the crown of winter wheat seedlings. Can J Plant Sci 70:1179–1182

    Article  Google Scholar 

  77. Pomeroy MK, Andrews CJ (1985) Effect of low temperature and calcium on survival and membrane properties of isolated winter wheat cells. Plant Physiol 78:484–488

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  78. Gudleifsson BE (1989) Laboratoriemetoder til testing av isdekketoleranse hjå gras. [Laboratory methods to study ice tolerance of grasses]. Nord Jordbruksforskn 71:81, In Norwegian

    Google Scholar 

  79. Björnsson H (1986) Statistisk analys och planläggning av laboratorieførsøk med frost- och isresistens. Statistical analysis and planning of laboratory testing of freezing and ice tolerance]. NJF seminar Nr. 84 Lantbruksväxternas övervintring, pp 176–179, [In Swedish]

    Google Scholar 

  80. Gudleifsson BE (1993) Methods for testing ice encasement tolerance in herbage plants. [Methods for testing ice tolerance of herbage plants]. Röbäcksdalen Meddelar 1993:83–94, In Norwegian

    Google Scholar 

  81. Rakitina ZG (1977) Effect of a surrounding ice crust on winter wheat plant as a function of their flooding before freezing. Sov Plant Physiol 24:317–324

    Google Scholar 

  82. Olien CR, Smith MN (1981) Extension of localized freeze injury in barley by acute post-thaw bacterial disease. Cryobiology 18:404–409

    Article  CAS  PubMed  Google Scholar 

  83. Gudleifsson BE (1983) Isdekkeresistens og frostherdsle hjå enggras. [Ice tolerance and frost tolerance of meadow grasses]. Nord Jordbruksforskn 65, 3, [In Norwegian]

    Google Scholar 

  84. Gudleifsson BE (1997) Microbes active under snow and ice cover in hayfields in Iceland. In: International workshop on plant-microbe interactions at low temperatures under snow. Hokkaido National Agricultural Experiment Station, Hitujioaoka, pp 109–118

    Google Scholar 

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Gudleifsson, B.E., Bjarnadottir, B. (2014). Estimating Ice Encasement Tolerance of Herbage Plants. In: Hincha, D., Zuther, E. (eds) Plant Cold Acclimation. Methods in Molecular Biology, vol 1166. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-0844-8_17

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  • DOI: https://doi.org/10.1007/978-1-4939-0844-8_17

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