A comparison of analytical approaches for assessing freezing damage in black spruce using electrolyte leakage methods

1996 ◽  
Vol 74 (6) ◽  
pp. 952-958 ◽  
Author(s):  
K. D. Odlum ◽  
T. J. Blake
Keyword(s):  
Critical Temperature ◽  
Electrolyte Leakage ◽  
Black Spruce ◽  
Frost Hardiness ◽  
Initial Increase ◽  
Cool Temperature ◽  
Freezing Damage ◽  
Intact Seedling ◽  
Temperature Hardening ◽  
Analytical Approaches

To compare different methods of quantifying shoot frost damage during controlled plant freezing tests, frost hardening of black spruce (Picea mariana (Mill.) BSP) seedlings exposed to three temperature hardening regimes over 16 weeks was assessed using electrolyte leakage and intact seedling methods. Electrolyte leakage was expressed as index of injury and was quantified either as the temperature needed to induce an index of injury of 5% (DT5) or as the critical temperature (CT), the mildest temperature at which damage was first detected statistically. Damage to intact shoots was expressed as percent shoot browning and was quantified as the temperature at which 50% of needle tissue on the shoots was damaged (sLT50) or as the temperature at which 50% of terminal buds were killed (bLT50). Seedling response to hardening temperature varied, depending on the method used to quantify frost hardiness. When expressed as critical temperature, hardening continued over the 16 weeks at a constant rate with no differences detected between treatments. Intact seedling shoot damage, sLT50 and bLT50, described a hardening process in which there was a large initial increase in hardening in the first 8 weeks, with less hardening occurring during the subsequent 8 weeks. Also, significant temperature effects were detected, with the greatest hardening occurring in a cool temperature (4 °C), the least in a warm temperature (20 °C), and an intermediate amount in a moderate temperature (10 °C). When quantified as DT5, the pattern of hardening was somewhat intermediate to the other two. Methods of determining frost hardiness were highly correlated, with the strongest correlation being between sLT50 and bLT50 (r2 = 0.903). Both electrolyte leakage methods, DT5 and CT, were linearly related to one another (r2 = 0.666) and were more sensitive than the intact seedling methods, since they both detected damage at warmer temperatures. DT5 was better correlated to intact measures of hardiness than was CT. Keywords: black spruce, index of injury, frost hardiness, critical temperature, damaging temperature, LT50.

Viability tests for estimating root cold tolerance of black spruce seedlings

1994 ◽  
Vol 24 (5) ◽  
pp. 1039-1048 ◽  
Author(s):  
Francine J. Blgras ◽  
Sophie Calmé
Keyword(s):  
Water Loss ◽  
Tissue Damage ◽  
Electrolyte Leakage ◽  
Fine Roots ◽  
Black Spruce ◽  
Root Systems ◽  
Test Results ◽  
Freezing Damage ◽  
Reactive Compound ◽  
Freezing Test

After freezing tests, various methods for evaluating root damage were compared and correlated to the regrowth of 16-week-old containerized black spruce seedlings (Piceamariana (Mill.) B.S.P.) hardened in growth chambers. During the hardening regime, three experiments were performed. The first experiment evaluated the influence on the test results of washing the roots before or after a freezing test and compared the frost sensitivity of fine roots (<0.3 mm), larger roots (>0.3 mm), and whole root systems. The second experiment tested the effect of pressures of 0.05, 0.10, and 0.15 MPa on water loss. Experiment 3 compared four viability tests: electrolyte leakage, ninhydrin reactive compounds leakage, phenolic leakage, and water loss. Results were as follows: (i) washing the roots before or after the freezing test did not affect the results of the ninhydrin reactive compounds test; (ii) fine roots (<0.3 mm) were more sensitive to freezing damage than larger roots (>0.3 mm); (iii) sampling of fine roots (<0.3 mm), larger roots (>0.3 mm), or whole root systems gave different results for the ninhydrin reactive compounds test; (iv) tests of electrolyte leakage, phenolic leakage, and ninhydrin reactive compounds leakage had the ability to indicate tissue damage; (v) high values of these variables were associated with poor seedling regrowth; (vi) root water potential also indicates tissue damage but is less correlated to regrowth than electrolyte leakage, phenolic leakage, and ninhydrin reactive compound leakage; and (vii) the water-loss test seems less reliable than the others.

Photoperiod influences dehardening of Chamaecyparisnootkatensis seedlings

1993 ◽  
Vol 23 (11) ◽  
pp. 2452-2454 ◽  
Author(s):  
B.J. Hawkins ◽  
S.E. McDonald
Keyword(s):  
Significant Influence ◽  
Electrolyte Leakage ◽  
Frost Hardiness ◽  
Controlled Environment

In early January, seedlings of yellow cypress (Chamaecyparisnootkatensis (D. Don) Spach) were placed in controlled-environment chambers under constant, increasing, and decreasing photoperiods. Seedlings from all treatments were assessed for frost hardiness at 2-week intervals using the freeze-induced electrolyte leakage method. Seedlings subject to increasing day lengths began to deharden immediately and at a greater rate than seedlings under a constant photoperiod. Seedlings in the decreasing photoperiod treatment maintained maximum hardiness for 42 days and then began to deharden spontaneously, although photoperiod continued to decrease. Once dehardening began in this treatment, it proceeded at the fastest rate of all treatments. These results indicate that photoperiod has a significant influence on the initiation and rate of dehardening in yellow cypress, and maximum hardiness cannot be maintained indefinitely.

Atmospheric CO2 enrichment and the development of frost hardiness in containerized black spruce seedlings

1990 ◽  
Vol 20 (9) ◽  
pp. 1392-1398 ◽  
Author(s):  
Hank A. Margolis ◽  
Louis-P. Vézina
Keyword(s):  
Black Spruce ◽  
Co2 Enrichment ◽  
Growing Season ◽  
Frost Damage ◽  
Development Stage ◽  
Frost Hardiness ◽  
Mineral Nutrient ◽  
Experimental Conditions ◽  
Frost Injury ◽  
Controlled Freezing

The hypothesis that a relatively brief exposure to elevated atmospheric CO2 could increase the frost resistance of shoots was tested on containerized black spruce seedlings (Piceamariana (Mill.) B.S.P.). Seedlings were exposed to 1000 ppm CO2 toward the end of their second growing season in an unheated production tunnel and in a heated greenhouse. In 1987, continuous 10-week CO2 exposures were applied in conjunction with mineral nutrient fertilization, and freezing tests were conducted each week. In 1988, a series of shorter 2-week CO2 exposures was applied to different groups of seedlings and no mineral nutrients were added. Controlled freezing tests were conducted at −10 °C and were followed by electrolytic conductivity measurements to assess frost injury. Under all experimental conditions, freezing tests on seedlings from both the production tunnel and the greenhouse indicated significantly greater frost damage for the CO2-enriched seedlings than for the controls. Late-growing season CO2 enrichment negatively affected the bud initiation – bud development stage of frost-hardiness development.

scholarly journals Effects of winter warming on cold hardiness and spring budbreak of four boreal conifers

Botany ◽  
2016 ◽  
Vol 94 (2) ◽  
pp. 117-126 ◽  
Author(s):  
Rongzhou Man ◽  
Steve Colombo ◽  
Pengxin Lu ◽  
Qing-Lai Dang
Keyword(s):  
Picea Glauca ◽  
Pinus Banksiana ◽  
Cold Hardiness ◽  
Black Spruce ◽  
Lodgepole Pine ◽  
Temperature Fluctuations ◽  
White Spruce ◽  
Jack Pine ◽  
Experimental Warming ◽  
Freezing Damage

Compared with the effects of spring frosts on opening buds or newly flushed tissues, winter freezing damage to conifers, owing to temperature fluctuations prior to budbreak, is rare and less known. In this study, changes in cold hardiness (measured based on electrolyte leakage and needle damage) and spring budbreak were assessed to examine the responses of four boreal conifer species — black spruce (Picea mariana (Mill.) B.S.P.), white spruce (Picea glauca) (Moench) Voss), jack pine (Pinus banksiana Lamb.), and lodgepole pine (Pinus contorta Dougl. ex. Loud.) — to different durations of experimental warming (16 °C day to –2 °C night with a 10 h photoperiod, except for night temperatures during November warming (+2 °C)). Seedlings showed increased responses to warming from November to March, while the capacity to regain the cold hardiness lost to warming decreased during the same period. This suggests an increasing vulnerability of conifers to temperature fluctuations and freezing damage with the progress of chilling and dormancy release from fall to spring. Both lodgepole pine and jack pine initiated spring growth earlier and had greater responses to experimental warming in bud phenology than black spruce and white spruce, suggesting a greater potential risk of frost/freezing damage to pine trees in the spring.

Photoperiod and night frost influence the frost hardiness of Chamaecyparisnootkatensis clones

1993 ◽  
Vol 23 (7) ◽  
pp. 1408-1414 ◽  
Author(s):  
B.J. Hawkins
Keyword(s):  
Electrolyte Leakage ◽  
Frost Hardiness ◽  
Relative Influence ◽  
Controlled Environment ◽  
Rooted Cuttings ◽  
Subzero Temperatures ◽  
Controlled Environments

Frost hardiness in yellow cypress (Chamaecyparisnootkatensis (D. Don) Spach) was studied over the winter to determine the relative influence of photoperiod and subzero temperatures on the hardening and dehardening processes. Stecklings (rooted cuttings) of five clones from each of three provenances were grown outdoors and in two controlled-environment chambers at 10:5 °C day:night temperatures and 12- or 6-h photoperiods. Half of the plants in each photoperiod treatment were subject to a 4-h night frost, three times per week from December through to March. Frost hardiness was assessed at intervals using the freeze-induced electrolyte leakage method. Variability in frost hardiness was greater among clones within provenances than among provenances. Significant differences in hardiness among clones existed throughout the experiment; however, the ranking of clones by hardiness was not consistent. Stecklings in the 6-h photoperiod were consistently more hardy than their counterparts subjected to 12-h photoperiods. Stecklings placed outdoors had equivalent hardiness to those in the 6-h photoperiod until the advent of natural frosts in January. At this time, the outdoor trees were the most hardy. Night frost also significantly increased frost hardiness in stecklings in controlled environments. Stecklings in all treatments began to deharden between January and March. The outdoor plants dehardened most rapidly. Stecklings in the controlled environments dehardened slowly until the night-frost treatment ended, whereupon previously frozen plants dehardened more rapidly than their unfrozen counterparts.

Effect of population, environment, and maturation on the frost hardiness of yellow-cedar (Chamaecyparisnootkatensis)

1994 ◽  
Vol 24 (5) ◽  
pp. 945-953 ◽  
Author(s):  
B.J. Hawkins ◽  
J. Russell ◽  
R. Shortt
Keyword(s):  
Air Temperature ◽  
Electrolyte Leakage ◽  
High Elevation ◽  
Genetic Component ◽  
Frost Hardiness ◽  
Diverse Populations ◽  
Rooted Cuttings ◽  
Stock Types

Three 2-year-old experimental plantations of yellow-cedar (Chamaecyparisnootkatensis (D. Don) Spach.) seedlings and stecklings (rooted cuttings) from 4- and 12-year-old hedges were analyzed periodically for frost hardiness between September 1991 and May 1992. The plantations were at different elevations and latitudes, and seedlings and stecklings of three geographically diverse populations were represented. Frost hardiness was assessed using the freeze-induced electrolyte leakage method on upper, primary branches. Air temperature at the low elevation sites was monitored. Frost hardiness of all plants increased from September to the end of January, and then decreased. Plants grown at high elevation were consistently more hardy than those grown at low elevation; however, no obvious differences in hardiness between the two latitudes were evident. Seedlings and stecklings of parents from certain families and populations developed greater hardiness than stock of parents from other areas, showing that frost hardiness of yellow-cedar plants has a genetic component. The three stock types did not show any consistent, significant differences in hardiness; however, stecklings from 12- and 4-year-old hedges were, on average, slightly more hardy than seedlings.

scholarly journals Standardization of electrolyte leakage data and a novel liquid nitrogen control improve measurements of cold hardiness in woody tissue

2021 ◽  
Author(s):  
Alisson Pacheco Kovaleski ◽  
Jake J Grossman
Keyword(s):  
Thermal Analysis ◽  
Differential Thermal Analysis ◽  
Liquid Nitrogen ◽  
Plant Tissue ◽  
Electrolyte Leakage ◽  
Cold Hardiness ◽  
Nitrogen Control ◽  
Freezing Damage ◽  
Raw Data ◽  
Reliable Determination

Abstract Background. A variety of basic and applied research programs in plant biology require the accurate and reliable determination of plant tissue cold hardiness. Over the past 50 years, the electrolyte leakage method has emerged as a popular and practical method for quantifying the amount of damage inflicted on plant tissue by exposure to freezing temperatures. Numerous approaches for carrying out this method and analyzing the resultant data have emerged. These include multiple systems for standardizing and modeling raw electrolyte leakage data and multiple protocols for boiling samples in order to maximize leakage as a positive control. We compare four different routines for standardization of leakage data and assess a novel control method - immersion in liquid nitrogen in lieu of traditional boiling – and apply them to woody twigs collected from 12 maple (Acer) species in early spring. We compare leakage data from these samples using each of four previously published forms of data analysis and boiling vs. liquid nitrogen controls and validate each of these approaches against visual estimates of freezing damage and differential thermal analysis. Results. Through presentation of our own data and re-analysis of previously published findings, we show that standardization of raw data against estimates of both minimum and maximum attainable freezing damage allows for reliable estimation of cold hardiness at the species level and across studies in diverse systems. Furthermore, use of our novel liquid nitrogen control produces data commensurate across studies and enhances the consistency and realism of the electrolyte leakage method, especially for very cold hardy samples. Conclusion. Future leakage studies that relativize data against minimum and maximum leakage and that employ our updated liquid nitrogen control will contribute generalizable, repeatable, and realistic data to the existing body of cold hardiness research in woody plants. Data from studies conducted using a liquid nitrogen (and not a boiling) control can still be compared to previously published data, especially when raw data are standardized using the best-performing approach among those we assessed. Electrolyte leakage of woody twigs emerges as a useful technique for quickly assessing the probability of tissue death in response to freezing in dormant plants. Differential thermal analysis may provide different and complementary information on cold hardiness.

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