New Forests

, 32:307 | Cite as

Overwinter Storability of Conifer Planting Stock: Operational Testing of Fall Frost Hardiness

  • Sylvia J. L’Hirondelle
  • David G. Simpson
  • Wolfgang D. Binder
Original Article


Operational stock-testing facilities that estimate overwinter storability of seedlings (ability to survive and grow after storage) need a reliable method that provides fast results to forest nurseries. We compared three methods using container-grown seedlings of Douglas-fir, interior spruce, lodgepole pine, and western larch from forest nurseries in British Columbia. On three to nine dates in autumn, frost hardiness at −18°C was estimated using visible injury of foliage or stems (VI), electrolyte leakage from needles or stems (EL), and chlorophyll fluorescence of shoots (CF). Seedlings were placed into overwinter cold storage (−2°C). In the spring, stored seedlings were planted in nursery beds; survival and growth were assessed after one growing season. There were close correlations (r ≥ 0.93) between the assessment methods. Seedlings lifted after they reached thresholds of 69% or higher for CF and 25% or lower for EL and VI had over 90% survival at harvest and doubled shoot dry weight compared with seedlings lifted earlier. Measuring CF was the fastest and most easily replicated method to estimate successful storability, and reduced testing time by 6 days relative to VI tests.

Key words

Chlorophyll fluorescence Electrolyte leakage Visible injury Freeze-testing 



We wish to thank current and former Ministry of Forests staff for their advice, support and encouragement at various stages of this project: Jim Sweeten, Clare Kooistra, Kendel Thomas, Ralph Huber, Shon Ostafew, Hazel Ritchie, B.J. Finnson, Wendy Clarke, Chris Hawkins, Glen Ursel, and Drew Brazier. We also thank two anonymous reviewers for their helpful suggestions.


  1. Adams GT, Perkins TD (1993) Assessing cold tolerance in Picea using chlorophyll fluorescence. Environ Exp Bot 33:377–382CrossRefGoogle Scholar
  2. Aronsson A, Eliasson L (1970) Frost hardiness in Scots pine (Pinus silvestris L.). I. Conditions for test on hardy plant tissues and for evaluation of injuries by conductivity measurements. Stud For Suec 77:1–30Google Scholar
  3. Bartlett KJ (2000) Just the facts: a review of silviculture and other forestry statistics. Report No. BC Ministry of Forests, 125 ppGoogle Scholar
  4. Binder WD, Fielder P (1996a) Seasonal changes in chlorophyll fluorescence of white spruce seedlings from different latitudes in relation to gas exchange and winter storability. New For 11:207–232Google Scholar
  5. Binder WD, Fielder P (1996b) Chlorophyll fluorescence as an indicator of frost hardiness in white spruce seedlings from different latitudes. New For 11:233–253Google Scholar
  6. Binder WD, Fielder P, Mohammed GH, L’Hirondelle SJ (1997) Applications of chlorophyll fluorescence for stock quality assessment with different types of fluorometers. New For 13:63–89Google Scholar
  7. Burdett AN, Simpson DG (1984) Lifting, grading, packaging and storing. In: Duryea ML, Landis TD (eds), Forest nursery manual: Production of bareroot seedlings. Martinus Nijhoff/Dr. W. Junk, The Hague, pp 227–234Google Scholar
  8. Burr KE, Tinus RW, Wallner SJ, King RM (1990) Comparison of three cold hardiness tests for conifer seedlings. Tree Physiol 6:351–369PubMedGoogle Scholar
  9. Burr KE, Hawkins CDB, L’Hirondelle SJ, Binder WD, George MF, Repo T (2001) Methods for measuring cold hardiness of conifers. In: Bigras F, Colombo S (eds), Conifer cold hardiness. Kluwer Academic Press, Dordrecht, Netherlands, pp 369–401Google Scholar
  10. Calkins JB, Swanson BT (1990) The distinction between living and dead plant tissue – viability tests in cold hardiness research. Cryobiology 27:194–211CrossRefGoogle Scholar
  11. Camm EL, Harper GJ, Rosenthal SI, Camm DM (1993) Effect of photon flux density on carbon assimilation and chlorophyll a fluorescence of cold-stored white spruce and lodgepole pine seedlings. Tree Physiol 12:185–194PubMedGoogle Scholar
  12. Camm EL, Goetze DC, Silim SN, Lavender DP (1994) Cold storage of conifer seedlings: an update from the British Columbia perspective. For Chron 70:311–316Google Scholar
  13. Chomba BM, Guy RD, Weger HG (1993) Carbohydrate reserve accumulation and depletion in Engelmann spruce (Picea engelmannii Parry): effects of cold storage and pre-storage CO2 enrichment. Tree Physiol 13:351–364PubMedGoogle Scholar
  14. Colombo SJ, Webb DP, Glerum C (1984) Frost hardiness testing: an operational manual for use with extended greenhouse culture. Forest Research Report No. 110. Ontario Ministry of Natural Resources, Ontario Tree Improvement and Forest Biomass Institute, 14 ppGoogle Scholar
  15. Colombo SJ, Raitanen EM (1993) Frost hardening in first-year eastern larch (Larix laricina) container seedlings. New For 7:55–61Google Scholar
  16. Colombo SJ (1997) Frost hardening spruce container stock for overwintering in Ontario. New For 13:449–467Google Scholar
  17. Damesin C (2003) Respiration and photosynthesis characteristics of current-year stems of Fagus sylvatica: from the seasonal pattern to an annual balance. New Phytol 158:465–475CrossRefGoogle Scholar
  18. Dexter ST, Tottingham WE, Graber LF (1932) Investigations of the hardiness of plants by measurement of electrical conductivity. Plant Physiol 7:63–78PubMedGoogle Scholar
  19. Fisker SE, Rose R, Haase DL (1995) Chlorophyll fluorescence as a measure of cold hardiness and freezing stress in 1 + 1 Douglas-fir seedlings. Forensic Sci 41:564–575Google Scholar
  20. Gillies SL, Vidaver WE (1991) The effect of pre-lifting temperature on photosynthesis and post-storage recovery in conifers. In: Donnelly FP, Lussenburg HW (eds) Forest Nursery Association of British Columbia Meeting. Forest Nursery Association of British Columbia, Prince George, BC, pp 15–21Google Scholar
  21. Hawkins CDB, Binder WD (1990) State of the art seedling stock quality tests based on seedling physiology. In: Rose R, Campbell SJ, Landis TD (eds) Target Seedling Symposium: Combined Meeting of the Western Forest Nursery Associations, Roseburg, Oregon, Aug. 13–17, USDA Forest Service, Rocky Mountain Forest and Range Experiment Station, Fort Collins CO, General Technical Report RM-200, pp 91–121Google Scholar
  22. Jenkinson JL, Nelson JA (1984) Cold storage increases resistance to dehydration stress in Pacific Douglas-fir. In: (eds) Western Forest Nursery Council and Intermountain Nurseryman’s Association Meeting, Coeur d’Alene, ID, USDA Forest Service, pp 38–44Google Scholar
  23. Jiang Y, Zwiazek JJ, MacDonald SE (1994) Effects of prolonged cold storage on carbohydrate and protein content and field performance of white spruce bareroot seedlings. Can J For Res 24:1369–1375CrossRefGoogle Scholar
  24. Jiang Y, MacDonald SE, Zwiazek JJ (1995) Effects of cold storage and water stress on water relations and gas exchange of white spruce (Picea glauca) seedlings. Tree Physiol 15:267–273PubMedGoogle Scholar
  25. Kooistra CM (2004) Seedling storage and handling in western Canada. In: Riley LE, Dumroese RK, Landis TD (eds) National Proceedings: Forest and Conservation Nursery Associations – 2003. Coeur d’Alene, ID, USDA Forest Service, Rocky Mountain Research Station, RMRS-P-33, pp 15–21Google Scholar
  26. Lindgren K, Hällgren J-E (1993) Cold acclimation of Pinus contorta and Pinus sylvestris assessed by chlorophyll fluorescence. Tree Physiol 13:97–106PubMedGoogle Scholar
  27. Luoranen J, Repo T, Lappi J (2004) Assessment of the frost hardiness of shoots of silver birch (Betula pendula) seedlings with and without controlled exposure to freezing. Can J For Res 34:1108–1118CrossRefGoogle Scholar
  28. Manetas Y (2004) Probing corticular photosynthesis through in vivo chlorophyll fluorescence measurements: evidence that high internal CO2 levels suppress electron flow and increase the risk of photoinhibition. Physiol Plant 120:509–517PubMedCrossRefGoogle Scholar
  29. Mattsson A (1997) Predicting field performance using seedling quality assessment. New For 13:227–252Google Scholar
  30. Maxwell K, Johnson GN (2000) Chlorophyll fluorescence – a practical guide. J Exp Bot 51:659–668PubMedCrossRefGoogle Scholar
  31. McKay HM, Mason WL (1991) Physiological indicators of tolerance to cold storage in Sitka spruce and Douglas-fir seedlings. Can J For Res 21:890–901Google Scholar
  32. McKay HM (1994) Frost hardiness and cold-storage tolerance of the root system of Picea sitchensis, Pseudotsuga menziesii, Larix kaempferi, and Pinus sylvestris bare-root seedlings. Scand J For Res 9:203–213Google Scholar
  33. Mohammed GH, Binder WD, Gillies S (1995) Chlorophyll fluorescence: a review of its practical forestry applications and instrumentation. Scand J For Res 10:383–410CrossRefGoogle Scholar
  34. Neuner G, Buchner O (1999) Assessment of foliar frost damage: a comparison of in vivo chlorophyll fluorescence with other viability tests. Angew Bot 73:50–54Google Scholar
  35. Nilsson H-E (1995) Remote sensing and image analysis in plant pathology. Annu Rev Phytopath. 33:489–527CrossRefGoogle Scholar
  36. O’Reilly C, McCarthy N, Keane M, Harper CP (2000) Proposed dates for lifting Sitka spruce planting stock for fresh planting or cold storage, based on physiological indicators. New For 19:117–141Google Scholar
  37. O’Reilly C, Harper CP, McCarthy N, Keane M (2001) Seasonal changes in physiological status, cold storage tolerance and field performance of hybrid larch seedlings in Ireland. Forestry 74:407–421CrossRefGoogle Scholar
  38. Perks MP, Monaghan S, O’Reilly C, Osborne BA, Mitchell DT (2001) Chlorophyll fluorescence characteristics, performance and survival of freshly lifted and cold stored Douglas fir seedlings. Ann For Sci 58:225–236CrossRefGoogle Scholar
  39. Pulkkinen P (1993) Frost hardiness development and lignification of young Norway spruce seedlings of southern and northern Finnish origin. Silva Fenn 27:47–54Google Scholar
  40. Ritchie GA (1987) Some effects of cold storage on seedling physiology. Tree Planters’ Notes 38:11–15Google Scholar
  41. Ritchie GA (1991) Measuring cold hardiness. In: Lassoie JP, Hinckley TM (eds) Techniques and approaches in forest tree ecophysiology. CRC Press, Boca Raton, FL, pp 557–582Google Scholar
  42. Ritchie GA (2004) Container seedling storage and handling in the Pacific Northwest: answers to some frequently asked questions. In: Riley LE, Dumroese RK, Landis TD (eds) National Proceedings: Forest and Conservation Nursery Associations – 2003, Coeur d’Alene, ID, USDA Forest Service, Rocky Mountain Research Station, RMRS-P-33, pp 3–7Google Scholar
  43. Rose R, Haase D (2002) Chlorophyll fluorescence and variations in tissue cold hardiness in response to freezing stress in Douglas-fir seedlings. New For 23:81–96Google Scholar
  44. Rosenthal SI, Camm EL (1997) Photosynthetic decline and pigment loss during autumn foliar senescence in western larch (Larix occidentalis). Tree Physiol 17:767–775PubMedGoogle Scholar
  45. Simpson DG (1990) Frost hardiness, root growth capacity, and field performance relationships in interior spruce, lodgepole pine, Douglas-fir, and western hemlock seedlings. Can J For Res 20:566–572CrossRefGoogle Scholar
  46. Steponkus PL (1984) Role of the plasma membrane in freezing injury and cold acclimation. Annu Rev Plant Physiol 35:543–584CrossRefGoogle Scholar
  47. Stergios BG, Howell GS, Jr (1973) Evaluation of viability tests for cold stressed plants. J Am Soc Hort Sci 98:325–330Google Scholar
  48. Tinus RW (1996) Cold hardiness testing to time lifting and packing of container stock: a case history. Tree Planters’ Notes 47:62–67Google Scholar
  49. Tinus RW, Burr KE (1997) Cold hardiness measurement to time fall lifting. In: Landis TD, Thompson JR (eds), National Proceedings, Forest and Conservation Nursery Associations, USDA Forest Service, Pacific Northwest Research Station, Portland, OR, Gen. Tech. Rep. PNW-GTR-419, pp 17–22Google Scholar
  50. Wang Y, Zwiazek JJ (1999) Effects of storage temperature on physiological characteristics of fall-lifted white spruce (Picea glauca) bareroot seedlings. Can J For Res 29:679–686CrossRefGoogle Scholar
  51. Westin J, Sundblad L-G, Hällgren J-E (1995) Seasonal variation in photochemical activity and hardiness in clones of Norway spruce (Picea abies). Tree Physiol 15:685–689PubMedGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2006

Authors and Affiliations

  • Sylvia J. L’Hirondelle
    • 1
  • David G. Simpson
    • 2
  • Wolfgang D. Binder
    • 1
  1. 1.Research Branch LaboratoryBC Ministry of ForestsVictoriaCanada
  2. 2.Kalamalka Forestry CentreBC Ministry of ForestsVernonCanada

Personalised recommendations