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  • Author or Editor: Michele R. Warmund x
  • Journal of the American Society for Horticultural Science x
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`Earliglow' strawberry (Fragaria xananassa Duchesne) plants were frozen to -5 or -50C to examine the distribution of ice in the crowns. Anatomical studies were also performed to characterize tissue growth in a greenhouse at 4, 8, and 15 weeks after freezing to -5C. Ice masses observed in fresh crown tissue corresponded to the presence of extracellular tissue voids in specimens fixed for scanning electron microscopy (SEM). Voids were present near the peduncle and adjacent to the vascular system in crown tissue. After plants were grown in the greenhouse, cell division and enlargement were observed near the voids in crowns subjected to -5C. By 15 weeks after freezing, a few small extracellular voids remained in the crowns. Tissue voids were also present in crowns of plants frozen rapidly to -50C and subsequently thawed. Cells in the crown of these plants were intact and did not appear collapsed after exposure to -50C, a lethal temperature.

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INA bacteria were isolated from primary flowers of `Totem' strawberry (Fragaria ×ananassa Duch.) plants that had been previously inoculated with strain Cit 7 of Pseudomonas syringae van Hall or noninoculated to determine their relationship to ice-nucleation temperature and floral injury. Mean ice-nucleation temperature of inoculated and noninoculated flowers was -2.2 and -2.8 °C, respectively. Primary flowers of noninoculated plants survived lower temperatures than those of inoculated plants. In another experiment, noninoculated plants were misted with sterile deionized water and incubated for 0, 12, 24, 36, or 48 hours at 25 °C day/10 °C night, and naturally occurring INA bacteria were isolated from primary flowers. INA bacterial densities increased exponentially with increasing incubation period. The critical wetness period for INA bacteria to establish a sufficient density to increase the likelihood of floral injury at -2.5 °C was 24 hours. Longer wetness periods resulted in higher INA bacterial densities but did not increase the floral mortality rate. Thermal analysis demonstrated that the ice nucleation temperature was associated with strawberry floral injury. Thus, low temperature survival of flowers was adversely affected by moisture for ≥24 h due to the presence of a sufficient density of INA bacteria to incite ice formation and floral injury.

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In 1993, ice-nucleation-active (INA) bacteria were isolated from `Redwing' red raspberries (Rubus idaeus L. var. idaeus) at five pigmentation stages. Fruit were also subjected to thermal analysis to determine the ice nucleation temperatures. INA bacteria were recovered from nearly all fruit samples, and the bacterial populations tended to decrease with greater red color development (i.e., fruit maturation). However, the ice nucleation temperature was not affected by the stage of fruit pigmentation. In 1994, INA bacterial densities were similar among fruit at the three pigmentation stages sampled. INA bacteria were recovered more often from the calyx rather than the drupe surface of these fruit. INA bacteria also were detected on pistils of some fruit. Red and pink fruit, which were nucleated with ice, had greater receptacle injury than mottled, yellow, or green fruit, but INA bacterial densities apparently were not related to injury. Thus, the injury response of fruit at different pigmentation (or development) stages indicated that nonbacterial ice nuclei may be involved in freezing injury of developing raspberries.

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Rhizomes of `Meyer' zoysiagrass (Zoysia japonica Steud.) were subjected to temperatures below 0 °C and were subsequently placed in a growth chamber with air at 34 °C day/28 °C night to determine the rate of shoot growth from nodes. Rhizomes exposed to subzero temperatures produced shoots steadily up to 16 days after freezing (DAF), but subsequent shoot growth from rhizomes was minimal. At 32 DAF, shoots were present on 68% and 44% of the nodes of unfrozen control (2 °C) rhizomes and those frozen to -7 °C, respectively. In another study, samples were frozen to a sublethal temperature (-7 °C) to examine the distribution of extracellular ice voids near the apical meristems of rhizomes and to characterize tissue recovery. Extracellular voids were present within the leaf tissue and between the leaves in samples prepared for scanning electron microscopy (SEM) immediately after freezing to -7 °C. By 12 DAF, most of the remaining voids were observed in older leaves. Nearly all extracellular voids in the leaves were absent by 20 DAF. However, by 28 DAF, some rhizomes still had small voids between leaves. Although the structure of zoysiagrass rhizomes subjected to -7 °C was temporarily disrupted, tissues recovered from extracellular freezing and new shoot growth was produced following exposure to warm temperatures.

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`Hull Thornless' and `Black Satin' blackberry (Rubus spp.) canes were collected from Sept. 1989 through Mar. 1990 to determine the hardiness and supercooling characteristics of buds at various stages of development. Anatomical studies were also conducted to examine the location of ice voids in buds frozen to -5 or -30C. Differentiation of the terminal flower occurred in `Black Satin' buds by 6 Nov., whereas `Hull Thornless' buds remained vegetative until early spring. As many as nine floral primordia were observed in both cultivars by 12 Mar. The hardiness of the two cultivars was similar until February. Thereafter, `Black Satin' buds were more susceptible to cold injury than those of `Hull Thornless'. Flora1 and undifferentiated buds of both cultivars exhibited one to four low temperature exotherms (LTEs) from 9 Oct. to 12 Mar. in differential thermal analysis (DTA) experiments. The stage of flora1 development did not influence the bud's capacity to supercool. The number of LTEs was not related to the stage of floral development or to the number of floral primordia. Extracellular voids resulting from ice formation in the bud axis and scales were observed in samples subjected to -5 or -30C.

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Abstract

Differential thermal analysis (DTA) experiments, viability tests, and anatomical studies were conducted to investigate the biophysics of freezing in ‘Darrow’ blackberry (Rubus spp.) buds at selected stages of development from Nov. 1985 through Sept. 1986. As many as four low-temperature exotherms (LTEs) associated with the crystallization of supercooled water were detected in DTA experiments on buds collected 16 Nov. 1985. Anatomical observations revealed seven to nine floral initials present at that date. On 16 Jan. 1986, buds were morphologically similar to those examined in November, with four to 10 LTEs per bud. By 2 Mar., the size of the floral initials increased and distinct floral parts were evident. One to 10 LTEs were observed per bud at this date. Five to seven floral initials were observed in buds collected on 23 Sept. 1986, but floral parts were not evident. DTA experiments conducted in September revealed one or no LTE per bud. The median LTEs for November, January, March, and September buds were −20.5°, −28.0°, −22.0°, and −16.5°C, respectively. T50 values calculated from viability tests were within 4.5° of the median LTEs at all test dates. These results indicate that injury to the entire floral region is associated with a single freezing event when ‘Darrow’ blackberry buds are at an early stage of development. However, as floral differentiation progresses, individual primordia supercool, freeze, and are injured independently.

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`Jonagold'/Mark apple (Malus domestica Borkh.) trees that were chip-budded in Washington and Illinois on 31 Aug. or 21 Sept. 1989 were sampled in Apr. 1990 to determine if magnetic resonance imaging (MRI) could be used to nondestructively examine vascular continuity or discontinuity between the rootstock and scion. Images could be placed into three categories based on signal intensity: 1) the rootstock, bud shield, and the bud or new scion growth had a high signal intensity; 2) the rootstock and the bud shield had a high signal intensity, but the scion had a low signal intensity; and 3) the rootstock had a high signal intensity, but the bud shield and scion had a low signal intensity. High signal intensity was associated with bound water in live tissue and the establishment of vascular continuity between the rootstock and scion. Azosulfamide staining and destructive sectioning confirmed that vascular continuity was established when the rootstock, bud shield, and scion had a high signal intensity in images, whereas budding failure occurred when the bud shield and/or the scion had a low signal intensity. Additional trees that had wilted or weak scion growth were collected from Illinois in June 1990. Parenchyma tissue was found in the scion adjacent to the bud shield that interrupted the vascular tissue. Poor scion growth on trees from the 21 Sept. budding in Washington may be attributed to insufficient growth of rootstock and/or scion tissues at the union in the fall.

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