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Courtney A. Weber, Gloria A. Moore, Zhanao Deng, and Fred G. Gmitter Jr.

Mapping quantitative trait loci (QTL) associated with freeze tolerance was accomplished using a Citrus grandis (L.) Osb. × Poncirus trifoliata (L.) Raf. F1 pseudo-testcross population. A progeny population of 442 plants was acclimated and exposed to temperatures of -9 °C and -15 °C in two separate freeze tests. A subpopulation of 99 progeny was genotyped for random amplified polymorphic DNA (RAPD), cleaved amplified polymorphic sequence (CAPS), sequence characterized amplified region (SCAR), and sequence tagged site (STS) markers to produce a linkage map for each parent. Potential QTL were identified by interval mapping, and their validity was corroborated with results from means comparison (t test), one-way analysis of variance (F test), and bulked segregant analysis (BSA). Multiple analytical methods provided evidence supporting putative QTL and decreased the probability of missing significant QTL associated with freeze tolerance. QTL with a large effect on freeze tolerance were located on both the Citrus and Poncirus linkage maps. In addition, clusters of markers with significantly different means between marker present and absent classes indicating minor QTL that contribute smaller effects on the level of tolerance were found on the linkage maps of both species.

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Jason D. Hinton, David P. Livingston III, Grady L. Miller, Charles H. Peacock, and Tan Tuong

excellent shoot growth similar to unfrozen controls. The plug green-up ratings were used to determine freeze tolerance or LT 50 , the temperature at which no regrowth occurs in 50% of the plants, through nonlinear regression using an equation adopted from

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Rita L. Hummel and Peter R. Bristow

In Spring 1996, `Meeker' red raspberry root cuttings were planted into a sandy loam soil in 30 cm tall x 27 cm diameter black plastic containers. During Mar. 1997, a second bottomless container was placed over the overwintering canes of half of the plants. The second container was filled with the same sandy loam soil to simulate ridging of the plants. All plants were grown using standard cultural practices on an outdoor, gravel nursery bed. Freeze tolerance of potted whole plants and excised root sections was measured at 5 °C intervals between -5 and -20 °C in a series of laboratory freeze tests conducted during Jan. 1998. Electrolyte leakage data were used to calculate the index of injury for excised roots while whole-plant response to freezing was determined by measuring the subsequent growth of floricane lateral shoots and of primocanes. After 1 month in the greenhouse, results indicated the dry weight of primocanes harvested from plants that were exposed to -20 °C was 56% of the nonfrozen control primocane dry weight. Primocane dry weight from plants exposed to -5, -10 and -15 °C was not different from the controls. Similar results were obtained for the percent of floricanes that were alive and for the dry weight of laterals produced by these floricanes after 3 months in the greenhouse. The whole-plant freeze test results indicated plants at the lowest temperature, -20 °C, were injured but not killed. Root index of injury of single potted plants averaged 5%, 15%, 29%, and 58% at -5, -10, -15, and -20 °C, respectively.

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D.O. Ketchie and R. Kammereck

Differential thermal analysis (DTA) and tetrazolium triphenyl chloride (TTC) were done on shoots of 4-year-old `Braeburn' apple trees for 3 years. The trees acclimated slowly in autumn. If cold temperatures last long enough in winter, shoots will acclimate as low as –40C. Shoots are sensitive to warm temperatures and deacclimated rapidly. An attempt to run a controlled test on freeze resistance of `Braeburn' did not respond to DTA. Moisture samples indicated trees were freeze dried. Different sets of trees were rehydrated and showed an exotherm pattern. Exotherms could be seen after 3 days at 26C, 14 days at 10C, and 21 days at 4C. Another controlled freeze test was performed on 1-year-old `Braeburn' trees. Trees were acclimated outdoors. An exotherm pattern could be seen upon DTA analysis. After artificial freezing, DTA and TTC tests showed pith killed at –24C, primarily xylem at –28C, and all tissue at –35C. After freezing, trees were placed in a greenhouse and warmed over 2 months. Upon dissection, we found xylem produced before freezing was dead, but a large amount of new xylem was generated. Trees appeared to have normal leaf and shoot growth for about a month, but eventually wilted and died. Dissection of these showed the same results as the first set dissected. New xylem evidently was not enough to carry the growth of the trees.

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Jeff A. Anderson

contains hundreds of genes under the control of CBF (C-repeat) transcription factors ( Stockinger, 2009 ). COR proteins and products of additional low temperature-induced pathways act directly or indirectly to increase freeze tolerance. It appears that

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Jeffrey A. Anderson, Charles M. Taliaferro, and Dennis L. Martin

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Orville M. Lindstrom, Tomasz Anisko, and Michael A. Dirr

Although differential thermal analysis has been routinely used to evaluate cold hardiness, the relationship between deep supercooling ability and plant survival is not clear. We compared seasonal profiles of changes in low-temperature exotherm (LTE) occurrence and visually determined lowest survival temperature (LST) of Acer rubrum `Armstrong', Fraxinus americana `Autumn Purple' and Zelkova serrata `Green Village' growing in three locations representing plant cold hardiness zones 8, 7 and 5. Between December and February, LTE in Acer rubrum and Fraxinus americana occurred at temperatures 10 to 25C lower than the LST. The difference between LTE and LST was not significant for Zelkova serrata from January to April, and for Acer rubrum and Fraxinus americana in March. Data indicate that LTE could be used as an estimate of LST in Zelkova serrata but not in Acer rubrum and Fraxinus americana. This study demonstrated that LTE does not provide a reliable estimate of cold hardiness in all species that deep supercool.

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Rita L. Hummel and Patrick P. Moore

The roles of freeze avoidance and freeze tolerance in determining strawberry (Fragaria ×ananassa) flower freeze resistance were compared in laboratory freeze tests. Genotype, freezing point depression of expressed cell sap, and flower size were examined as potential sources of variation in freeze resistance. When ice was added as a nucleator to excised flowers, mean freeze damage was 97% at -3.0 °C, but in the absence of ice, flowers appeared to supercool and had only 15% damage at -4.0 °C. Without nucleation, cultivar differences in freeze damage were significant in three of four freezing temperatures, but the relative ranking of cultivar freeze damage was not consistent across temperatures. Cultivars that sustained the least amount of injury at -4 °C, were not necessarily the least injured at -7 °C. With an ice nucleator, damage occurred at warmer temperatures (-1.5 °C), but there was no relationship between percentage damage at -1.5 °C with nucleation and -4 °C without nucleation across cultivars. Freezing-point depression of expressed cell sap did not account for the variation in freeze resistance. In nucleated and nonnucleated treatments, larger flowers were more likely to be freeze damaged. Results of this research suggest that flowers of all cultivars are susceptible to freeze damage and survive spring frosts by freeze avoidance.

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Orville M. Lindstrom

Leyland cypress [×Cupressocyparis leylandii (A.B. Jacks. and Dallim.) Dallim. and A.B. Jacks.] plants were transplanted into the field monthly from Aug. 1989 through Mar. 1990, and laboratory cold-hardiness estimates of these transplants were obtained monthly for two winter seasons. Cold hardiness estimates obtained in Dec. 1989 and Jan. 1990 revealed that the Nov. and Dec. 1989 transplants were 6C less cold-hardy than those transplanted into the field earlier in the year. There was little difference in cold hardiness due to transplant date during Feb., Mar., and Apr. 1990. In the second year of the study, on the same transplants, cold hardiness varied among transplanting dates. In Dec. 1990 and Jan. 1991, those transplanted in Jan.-Mar. 1990 were up to 9C less cold-hardy than those transplanted earlier in the season. However, in Mar. and Apr. 1991, those transplanted in Jan.-Mar. 1990 were equally or more cold-hardy than those transplanted earlier in the season. Transplanting Leyland cypress into the field in August to November appears to be the best time to ensure development of cold hardiness in early winter, whereas January to March planting appears to promote greater cold hardiness in the spring months.

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Orville M. Lindstrom

Whole, half, and quarter leaves and leaf disks were used to make laboratory estimations of the cold hardiness of Magnolia grandiflora. The effects of ice nucleation temperatures, length of exposure to nucleating temperatures, rates of temperature drop, thawing regimes, and methods of injury analysis were investigated for each leaf type in the fall and midwinter. In general, whole and half leaves responded more consistently to freezing tests than did quarter leaves and leaf disks. The most critical factors in the freezing procedure are the temperature at which the samples are nucleated with ice crystals and the regime in which the samples are warmed. These data suggest that whole and half leaves can effectively be used to reliably predict the cold hardiness of southern magnolia leaves.