Flower buds of two sweet cherry (Prunus avium L.), 12 sour cherry (Prunus cerasus L.) and one ground cherry (P. fruticosa Pall.) were collected monthly from Aug. 1990 to Mar. 1991, and subjected to freeze tests to determine the level of cold hardiness. LT50 values (temperatures at which 50% of the flower buds were killed) summed over all months were significantly correlated (r = 0.6844, P ≤ 0.01) to the flower bud low temperature exotherms (LTEs). Correlation of LTEs to LT50 values was highest, r = 0.85, P ≤ 0.01 for the acclimation and midwinter period, November to February collections. During this period the average LT50 occurred before and within 2.5 °C of the LTE, indicating tissue injury before the LTE occurrence. During deacclimation, represented by the March collection, the LT50 began within 2.0 °C, on average, of the LTE, and in 11 of 12 cultivars and seedlings preceded the LTE. In March, the correlation of LTEs to LT50 values was less, r = 0.69, P ≤ 0.05, indicating possible changes flower bud deep supercooling. LTE values were selected as a measure of flower bud hardiness in sour cherry. Exotherms were not detected in the flower buds of all germplasm tested on all evaluation dates, but were the best means of separating selections. While LTE analyses expressed significant differences in November, December, and March at P ≤ 0.01, the LT50 analyses expressed differences only in December and February at P ≤ 0.05. The relationship between ambient temperatures and floral tissue hardiness indicated that November and March are two critical times for flower bud injury. November injury would occur in years when sudden low temperatures occur without sufficient pre-exposure to freezing temperatures. March injury would occur in years when sudden freezing temperatures follow warm days. This type of injury would be most pronounced in southern genotypes. Spring freeze injury could be significantly reduced by the selection of cultivars and seedlings that have delayed deacclimation. Exotherm occurrence and bud volume were correlated (r = 0.95, P ≤ 0.05). In January, when exotherms were least prevalent, they were generally present only in the five cultivars and seedlings with large bud volumes. The LTEs in midwinter, occurred within 3 °C of the reported average annual minimum temperature for the northern range of Prunus commercial production (Zone 6). The results of the principal component analysis of flower bud LTEs indicated that other selection criteria as flowering time might have played a more significant role in the hardiness range of sour cherry than simply geographic origin. The first principal component (PC1), which accounted for 77% of the total variance was used to separate among cultivars and seedlings. Selections at the positive end of PC1 had flower buds that were more cold susceptible than selections at the negative end of PC. This concurs with other research showing that flower bud hardiness is related more to commercial range (i.e., the range of commercial production) than to geographic distribution.
Stephanie Byard, Michael Wisniewski, Jianhua Li, and Dale Karlson
subzero temperatures: freeze tolerance (nonsupercooling) and freezing avoidance (deep supercooling) ( Burke et al., 1976 ; George et al., 1982 ). In nonsupercooling cells, ice formation is initiated in extracellular spaces and intracellular water is
Cindy L. Flinn and Edward N. Ashworth
Thermal analysis of Forsythia × intermedia `Spectabilis' flower buds had previously detected the occurrence of low temperature exotherms (LTE) during freezing. The LTE apparently resulted from the freezing of supercooled water and corresponded to the death of the florets. The genus Forsythia encompasses a wide array of species and interspecific crosses ranging in flower bud hardiness and floret size. The ability of buds to supercool, the relationship between the LTE and flower bud hardiness, and the extent to which floret size affects both were studied in flower buds of the following Forsythia species: F. × intermedia `Spectabilis', F. × intermedia `Lynwood', F. `Meadowlark', F. suspensa var. fortunei, F. `Arnold Dwarf, F. europaea, F. giraldiana, F. × intermedia `Arnold Giant', F. japonica var. saxatilis, F. mandshurica, F. ovata, and F. viridissima. Flower buds used for thermal analysis were also used in subsequent size determinations. Hardiness evaluations were conducted using controlled freezing tests, and the sampling interval defined using the temperature range of the LTEs. Initial evaluation indicated a high degree of correlation (α>.50) between mean LTEs and mean killing temperatures. The Forsythia genus, with its broad range of bud hardiness and size provides an excellent system in which to study the mechanisms of supercooling. Thermal analysis of cultivars which exhibit LTEs can accurately assess bud hardiness with minimal plant material.
Sorkel A. Kadir and Ed L. Proebsting
Differential thermal analysis (DTA) was used to measure deep supercooling in flower buds of Prunus dulcis Mill., P. armeniaca L., P. davidiana (Carr.) Franch, P. persica (L.) Batsch, three sweet cherry (P. avium L.) selections, and `Bing' cherries (P. avium L.) during Winter 1990-91 and 1991-92. Low temperatures in Dec. 1990 killed many flower buds. After the freeze, dead flower primordia continued to produce low-temperature exotherms (LTEs) at temperatures near those of living primordia for >2 weeks. In Feb. 1992, cherry buds that had been killed by cooling to -33C again produced LTEs when refrozen the next day. As buds swelled, the median LTE (LTE50) of dead buds increased relative to that of living buds, and the number of dead buds that produced LTEs decreased. LTE artifacts from dead flower priimordia must be recognized when DTA is used to estimate LTE50 of field-collected samples.
Edward F. Durner and Thomas J. Gianfagna
The heat requirement for flower bud growth of container-grown peach trees [Prunus persica (L.) Batsch. cvs. Redhaven and Springold] in the greenhouse varied inversely and linearly with the length of the cold-storage period (SC) provided to break bud dormancy. Ethephon reduced the rest-breaking effectiveness of the 5C treatment. Buds from ethephon-treated trees grew more slowly than buds from untreated trees upon exposure to 20 to 25C, resulting in later bloom dates. The effect of ethephon on flower bud hardiness in field-grown trees of `Jerseydawn' and `Jerseyglo' was studied using exotherm analysis after deacclimation treatments. Bud deacclimation varied with reacclimating temperature (7 or 21 C), cultivar, ethephon treatment, and sampling date. All buds were more susceptible to injury in March than in January or February. Buds reacclimated more rapidly at 21C than at 7C. `Jerseyglo' reacclimated more rapidly than `Jerseydawn'. Untreated buds were less hardy and also reacclimated more rapidly than treated buds. Ethephon enhanced flower bud hardiness in three distinct ways: 1) it decreased the mean low-temperature exotherm of pistils, 2) it increased the number of buds that supercooled after exposure to reacclimating temperatures, and 3) it decreased the rate of deacclimation, especially at 21C. Ethephon prolongs flower bud dormancy by increasing the chilling requirement. The rate at which flower buds become increasingly sensitive to moderate temperatures in late winter and spring is thus reduced by ethephon. Thus, ethephon delays deacclimation during winter and delays bloom in the spring. Chemical name used: (2-chloroethyl) phosphoric acid (ethephon).
Knowledge of the level of cold hardiness and how hardiness is inherited in sour cherry is essential to germplasm collection and cultivar development. Twig samples of two sweet cherries (Prunus avium L.), 12 sour cherries (P. cerasus L.), and one ground cherry (P. fruticosa Pall.) of diverse geographic origins were collected in Jan. 1990 and monthly from Aug. 1990 to Mar. 1991, preconditioned to induce maximum cold resistance, and subjected to freeze tests and differential thermal analysis. Low temperature exotherms (LTEs) were detected in all stems of P. cerasus investigated and correlated to xylem incipient injury temperatures (ITs) from December to February (r = 0.84, P ≤ 0.01). March had the best correlation of LTEs to xylem ITs with r = 0.84, P ≤ 0.01. LTEs were strongly correlated to phloem-cambium ITs in November, representing the acclimation period. The correlation coefficient (r) for the phloem-cambium ITs and the twig LTEs during November was 0.68, significant at P ≤ 0.01. Cortical tissue and vegetative bud injuries were not correlated to the stem LTEs. Xylem ITs were selected for evaluating the cold resistance of sour cherry in December to March and phloem-cambium ITs were selected for November. The degree of supercooling and hardiness of the phloem-cambium in late fall and early spring appears significant in determining the stem hardiness and commercial range of P. cerasus. Phloem-cambium tissue, expressed the most rapid deacclimation response. The average decrease in hardiness for the phloem-cambium, xylem, and cortical tissues between February and March was 4 °C, 0.32 °C, and 2.14 °C, respectively. Principal component (PC) analyses of the phloem-cambium and cortical tissues depicted gradations between minimum survival temperatures of the two presumed progenitor species of sour cherry, i.e., sweet cherry and ground cherry. The first principal component (PC1), which accounted for 61% of the total variance, was used to separate among cultivars and seedlings. Cultivars and seedlings at the negative end of PC1 exhibited hardier phloem-cambium tissue at critical injury times, October, December, January, and March than cultivars and seedlings at the positive end of the PC1 axis. Cultivars and progeny of crosses of northern origin parents showed hardiness values more comparable to ground cherry than did selections of less-cold-hardy parents suggesting that cold is a major selective force, contributing to sour cherry population variation.
Jeffrey A. Anderson and Julia Whitworth
Frostgard did not effectively promote the supercooling of flowering `Arking' strawberry (Fragaria ×ananassa Duch.) plants in the presence or absence of ice-nucleation-active bacteria when applied as a spray in laboratory experiments. Frostgard effectively promoted supercooling and reduced the ice propagation rate of aqueous solutions. Detached leaves infiltrated with Frostgard exhibited a negative linear relationship between freezing temperature and Frostgard concentrations from 0% to 20% (by volume). Leaves infiltrated with 20% Frostgard supercooled 1.7C lower than those infiltrated with distilled water. Ice propagation barriers in strawberry plants were observed. Individual leaves froze independently, and a thermal ice propagation barrier sometimes was observed at the crown.
Michael Wisniewski and Glen Davis
The pit membrane of xylem parenchyma of peach plays an important role in deep supercooling. Enzyme hydrolysis of xylem tissue indicated that the pit membrane is rich in pectin. The objective of the present study was to determine if removal of calcium from the cell wall would effect deep supercooling by loosening the cell wall. Current year shoots of `Loring' peach were infiltrated with oxalic acid, EGTA, or sodium phosphate buffer for 24-48 hours and then prepared for either ultrastructural analysis or differential thermal analysis. The use of 5-50 mM oxalic acid resulted in a distinct reduction in the size of the low-temperature exotherm (LTE) with increasing concentration. Oxalic acid also produced a loosening and swelling of the pit membrane. The use of EGTA (100 mM) or NaP04 (150 mM) produced only a slight shift in the LTE to warmer temperatures when compared to fresh tissues. Heat treatments (30-100°C) also resulted in a gradual shift of the LTE to warmer temperatures. The data indicate that cross-linking of pectins may play a role in defining the pore structure of the pit membrane and that this area of the cell wall plays an integral role in deep supercooling of peach wood.
H.A. Quamme, Wei Ai Su, and L.J. Veto
Excision of the flower from the peach [Prunus persica (L.) Batsch.] flower bud raised the 50% injury temperature of flowers (cooled at 1C/hour) from -18 and -20C to -10 and -13C on two test dates, 26 Feb. 1988 and 5 Dec. 1990, respectively. Ice inoculation of the excised flowers at -3C further raised the 50% injury temperature to -7 and -8C for the two dates, respectively. These observations suggest that supercooling is a true mechanism for avoiding freezing injury. Low temperature scanning electron microscopy of freeze fractured cells verified that the flower froze intracellularly, whereas the subtending tissue froze extracellularly. Ice inoculation of the flower and the flower bud axis from which the scales were removed demonstrated that a barrier to ice propagation (effective to -11C) from the flower bud axis to the flower was present. This barrier may involve the provascular strands and a cell zone at the flower base (BZ) that were devoid of intercellular spaces. These tissues also had smaller cells, smaller vacuoles, greater ratio of cell wall thickness to cell size than tissue just below the BZ, which may result in greater cell rigidity and restriction of extracellular freezing. The cells outside the provascular strands at the base of the flower were also lacking in intercellular space, were smaller in size, and had a higher ratio of cell wall thickness to cell size compared to cells near the base of the scales. In the intact flower buds in which the flowers supercool below -11C, the presence of the first and second scales was important to full expression of supercooling because their removal raised the supercooling point, whereas the removal of lower scales did not. Sequestration of ice by the first two subtending scales during the early stages of freezing may be important to the creation of a dry region at the flower base that prevents ice propagation into the flower at temperatures below -11C.
Fanyi Shen, Rongfu Gao, Wenji Liu, Wenjie Zhang, and Qi Zhao
It is known that the redistribution of water and the formation of dispersed water units appears to be a prerequisite for deep supercooling. A concentration of the cell solute results from the migration of water during extracelullar freezing and lowers the temperature of homogeneous nucleation, but we are convinced that nucleation of ice within cells may be initiated by a heterogeneous mechanism, except we consider a small spherical cave, the water can freeze on the wall of this cave. We are also convinced that the solid walls of the capillary exert an external potential on the water molecules, causing the shift of the triple point of the confined fluids. Based on Fletcher's work for spherical particle, we have gotten the formula of critical free energy in the process of heterogeneous nucleation of water in a small spherical cave. This presentation introduces the theoretical background and counts the drop of temperature in heterogeneous nucleation. Then, putting two actions (depression of triple point and process of heterogeneous nucleation) together, we have calculated the freezing point. Sometimes it is lower than –38 °C. Some phenomena can be explained by using this theory: 1) Water is at the tension status, which means that it wets plant tissue, so the triple point (melting point) of tissue water can be lowered. 2) The redistribution of water, formation of dispersed water units, and dry region preventing ice from propagating, all allow heterogeneous nucleation, then the two actions can be synthesized and the water would lead to deep supercooling. If the barriers were destroyed, heterogeneous nucleation and deep supercooling would certainly be lost. 3) This theory is only suited to rigid wall of small cave, so we understand why cell wall rigidity has been shown to affect freezing characteristics. Project 39870234 supported by National Nature Science Foundation.