scholarly journals Ice Nucleation, Freezing Injury, and Colonization of `Totem' Strawberry Flowers with Ice-nucleation-active (INA) Bacteria

1998 ◽  
Vol 123 (2) ◽  
pp. 234-238 ◽  
Author(s):  
Michele R. Warmund ◽  
James T. English
Keyword(s):  
Pseudomonas Syringae ◽  
Ice Nucleation ◽  
Deionized Water ◽  
Freezing Injury ◽  
Nucleation Temperature ◽  
Ice Formation ◽  
Fragaria Ananassa ◽  
Naturally Occurring ◽  
Ina Bacteria ◽  
Wetness Period

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.

scholarly journals 557 PB 374 EFFICACIES OF CRYOPROTECTANTS APPLIED TO STRAWBERRY PLANTS INOCULATED WITH ICE-NUCLEATION-ACTIVE BACTERIA

HortScience ◽  
1994 ◽  
Vol 29 (5) ◽  
pp. 511d-511
Author(s):  
Michele R. Warmund ◽  
James T. English
Keyword(s):  
Low Temperature ◽  
Pseudomonas Syringae ◽  
Ice Nucleation ◽  
Freezing Injury ◽  
Fragaria Ananassa ◽  
Full Bloom ◽  
Ina Bacteria ◽  
Active Bacteria ◽  
Temperature Exposure ◽  
Low Temperature Exposure

Cryoprotectants were applied at labeled rates to primary flowers of `Honeoye' strawberry (Fragaria × ananassa Duch.) plants at full bloom to determine their effects on the floral organs. Frostgard at 50 ml/liter or KDL at 22 ml/liter injured pistils and resulted in misshapened fruit. Floral buds that were closed when cryoprotectants were applied were uninjured. In other experiments, efficacies of cryoprotectants were determined after floral tissues of `Honeoye' strawberry plants were inoculated or not inoculated with the ice-nucleation-active (INA) bacteria, Pseudomonas syringae van Hall and subjected to sub-freezing temperatures. None of the products protected primary or secondary flowers against freezing injury regardless of the occurrence of INA bacteria. INA bacteria were not recovered from primary flowers of treated plants that were killed by low temperature exposure, indicating that non-bacterial nuclei may incite freezing in these tissues.

scholarly journals Measurement of Ice Nucleation-Active Bacteria on Plants and in Precipitation by Quantitative PCR

2013 ◽  
Vol 80 (4) ◽  
pp. 1256-1267 ◽  
Author(s):  
Thomas C. J. Hill ◽  
Bruce F. Moffett ◽  
Paul J. DeMott ◽  
Dimitrios G. Georgakopoulos ◽  
William L. Stump ◽  
...  
Keyword(s):  
Quantitative Pcr ◽  
Pseudomonas Syringae ◽  
Xanthomonas Campestris ◽  
Ice Nucleation ◽  
Content Type ◽  
Biological Source ◽  
Ina Bacteria ◽  
A Minor ◽  
Ice Nucleation Activity ◽  
Effective Contribution

ABSTRACTIce nucleation-active (INA) bacteria may function as high-temperature ice-nucleating particles (INP) in clouds, but their effective contribution to atmospheric processes, i.e., their potential to trigger glaciation and precipitation, remains uncertain. We know little about their abundance on natural vegetation, factors that trigger their release, or persistence of their ice nucleation activity once airborne. To facilitate these investigations, we developed two quantitative PCR (qPCR) tests of theinagene to directly count INA bacteria in environmental samples. Each of two primer pairs amplified most alleles of theinagene and, taken together, they should amplify all known alleles. To aid primer design, we collected many new INA isolates. Alignment of their partialinasequences revealed new and deeply branching clades, including sequences fromPseudomonas syringaepv.atropurpurea,Ps. viridiflava,Pantoea agglomerans,Xanthomonas campestris, and possiblyPs. putida,Ps. auricularis, andPs. poae. qPCR of leaf washings recorded ∼108inagenes g−1fresh weight of foliage on cereals and 105to 107g−1on broadleaf crops. Much lower populations were found on most naturally occurring vegetation. In fresh snow,inagenes from various INA bacteria were detected in about half the samples but at abundances that could have accounted for only a minor proportion of INP at −10°C (assuming oneinagene per INA bacterium). Despite this, an apparent biological source contributed an average of ∼85% of INP active at −10°C in snow samples. In contrast, a thunderstorm hail sample contained 0.3 INA bacteria per INP active at −10°C, suggesting a significant contribution to this sample.

scholarly journals 993 THEORY AND APPLICATION OF GENETIC ENGINEERING FOR STRESS RESISTANCE AND AVOIDANCE

HortScience ◽  
1994 ◽  
Vol 29 (5) ◽  
pp. 571d-571
Author(s):  
Steven E. Lindow
Keyword(s):  
Population Size ◽  
Pseudomonas Syringae ◽  
Stress Resistance ◽  
Field Studies ◽  
Ice Nucleation ◽  
Site Directed Mutagenesis ◽  
Ice Formation ◽  
Bacterial Strains ◽  
Solanum Commersonii ◽  
Frost Injury

Genes determining the ability of the bacterium Pseudomonas syringae to catalyze ice formation have been cloned and characterized. Ice nucleation active (Ice+) strains of this species are common on plants and the supercooling ability of frost sensitive plants is inversely proportional to the logarithm of the population size of Ice+ bacteria at temperatures above -5C. Recombinant Ice- derivatives off. syringae were produced by site-directed mutagenesis using deletion containing ice genes cloned form this species. The Ice- strains colonized potatoes well in field studies, reduced the population size of Ice+ bacterial strains by about 50-fold, and reduced the incidence of frost injury an average of 82% in several radiative frosts of temperatures in the range of -3 to -5 C. The ice gene has also been introduced into Solanum commersonii to determine its effect on increasing the tolerance of ice formation in this frost tolerant species. Transgenic plants exhibit a much higher threshold ice nucleation temperature than the parental plants.

scholarly journals Characterization of Ice Nucleation and Propagation in Bean (Phaseolus vulgaris cv. Bush Blue Lake) using Infrared Video Thermography

HortScience ◽  
1997 ◽  
Vol 32 (3) ◽  
pp. 538C-538
Author(s):  
Michael Wisniewski
Keyword(s):  
Pseudomonas Syringae ◽  
Deionized Water ◽  
Ice Formation ◽  
Freezing Process ◽  
Blue Lake ◽  
Bean Plants ◽  
Infrared Video ◽  
Silicon Grease

Frost-sensitive plant species have a limited ability to tolerate ice formation in their tissues. Most plants can supercool below 0°C and avoid ice formation. Discrepancies exist about the role of intrinsic and extrinsic ice-nucleating agents in initiating ice formation in plants. Previous research has demonstrated the ability of infrared video thermography to directly observe and record the freezing process in plants (Wisniewski et al., 1997. Plant Physiol. 113:4378–4397). In the present study, the ability of droplets of a suspension of the ice-nucleating-active (Ice+) bacterium, Pseudomonas syringae, and droplets of deionized water, to induce ice formation in bean plants was compared. The activity of these agents were also compared to intrinsic ice formation in dry plants. Results indicated that the presence of the Ice+ bacteria in droplets ranging from 0.5–4.0 μL always induced freezing at a warmer temperature than droplets of deionized water alone (no bacteria) or intrinsic nucleators in dry plants. When droplets of Ice+ bacteria were allowed to dry, they were no longer effective but were active again upon rewetting. Droplets of water would often supercool below temperatures at which ice formation was initiated by intrinsic agents. When a silicon grease barrier was placed between the droplets of Ice+ bacteria and the leaf surface, the bacteria were no longer capable of inducing ice formation in the plant, despite the droplets being frozen on the plant surface. This indicates that ice crystals must penetrate the cuticle in order to induce freezing of the plant.

scholarly journals Ice-nucleation-active (INA) Bacteria: A Detriment to Strawberry Flower Survival during Low-temperature Exposure

HortScience ◽  
1997 ◽  
Vol 32 (3) ◽  
pp. 428B-428
Author(s):  
Michele R. Warmund ◽  
James T. English
Keyword(s):  
Low Temperature ◽  
Pseudomonas Syringae ◽  
Ice Nucleation ◽  
Ice Crystals ◽  
Colony Forming Units ◽  
Ina Bacteria ◽  
Temperature Exposure ◽  
Low Temperature Exposure ◽  
Ice Crystallization

Experiments were conducted to determine the temperatures at which different densities of INA bacteria incite ice crystallization on `Totem' strawberry flowers and to determine if there is a relationship between densities of INA bacteria on strawberry flowers and floral injury. Primary flowers were inoculated with Pseudomonas syringae at 106 cells/ml buffer, incubated at 25°C day/10°C night and 100% RH for 48 h, and exposed to –2.0°C. No ice nucleation occurred on these inoculated flowers and all of the flowers survived. However, when inoculated flowers were subjected to lower temperatures, ice nucleation occurred at –2.2°C and few of the flowers survived. In contrast, ice crystals formed on the surface of most non-inoculated flowers at –2.8°C and 21% of the flowers survived exposure to –3.5°C. When INA bacterial densities were ≈105 colony forming units/g dry wt, floral injury occurred at a warmer temperature than to flowers that had lower bacterial densities.

scholarly journals Effect of the expression of aquaporins 1 and 3 in mouse oocytes and compacted eight-cell embryos on the nucleation temperature for intracellular ice formation

Reproduction ◽  
2011 ◽  
Vol 142 (4) ◽  
pp. 505-515 ◽  
Author(s):  
Shinsuke Seki ◽  
Keisuke Edashige ◽  
Sakiko Wada ◽  
Peter Mazur
Keyword(s):  
Plasma Membrane ◽  
Plasma Membranes ◽  
Cell Types ◽  
Ice Nucleation ◽  
Nucleation Temperature ◽  
Ice Formation ◽  
Mouse Oocytes ◽  
Intracellular Ice Formation ◽  
Intracellular Ice ◽  
Mature Mouse

The occurrence of intracellular ice formation (IIF) is the most important factor determining whether cells survive a cryopreservation procedure. What is not clear is the mechanism or route by which an external ice crystal can traverse the plasma membrane and cause the heterogeneous nucleation of the supercooled solution within the cell. We have hypothesized that one route is through preexisting pores in aquaporin (AQP) proteins that span the plasma membranes of many cell types. Since the plasma membrane of mature mouse oocytes expresses little AQP, we compared the ice nucleation temperature of native oocytes with that of oocytes induced to express AQP1 and AQP3. The oocytes were suspended in 1.0 M ethylene glycol in PBS for 15 min, cooled in a Linkam cryostage to −7.0 °C, induced to freeze externally, and finally cooled at 20 °C/min to −70 °C. IIF that occurred during the 20 °C/min cooling is manifested by abrupt black flashing. The mean IIF temperatures for native oocytes, for oocytes sham injected with water, for oocytes expressing AQP1, and for those expressing AQP3 were −34, −40, −35, and −25 °C respectively. The fact that the ice nucleation temperature of oocytes expressing AQP3 was 10–15 °C higher than the others is consistent with our hypothesis. AQP3 pores can supposedly be closed by low pH or by treatment with double-strandedAqp3RNA. However, when morulae were subjected to such treatments, the IIF temperature still remained high. A possible explanation is suggested.

scholarly journals Freezing Injury and Colonization of `Redwing' Red Raspberries with-Ice-nucleation-active Bacteria

1995 ◽  
Vol 120 (6) ◽  
pp. 1041-1044
Author(s):  
Michele R. Warmund ◽  
James T. English
Keyword(s):  
Ice Nucleation ◽  
Rubus Idaeus ◽  
Freezing Injury ◽  
Fruit Maturation ◽  
Bacterial Populations ◽  
Color Development ◽  
Ice Nuclei ◽  
Red Color ◽  
Fruit Samples ◽  
Ina Bacteria

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.

scholarly journals JENIS DAN POPULASI BAKTERI ICE NUCLEATION ACTIVE PENYEBAB LUKA BEKU PADA DAUN JERUK KEPROK SOE DI DATARAN TINGGI MUTIS

2020 ◽  
Vol 3 (2) ◽  
pp. 127-135
Author(s):  
Hildegardis Missa ◽  
Anselmus Boy Baunsele
Keyword(s):  
Bacterial Population ◽  
Ice Nucleation ◽  
Test Method ◽  
Ice Formation ◽  
Frost Injury ◽  
Class B ◽  
Ina Bacteria ◽  
Three Stages ◽  
Active Bacteria ◽  
Ice Nucleation Activity

Research on ice nucleation-active bacteria causes frost injury from tropic areas has not been widely publicized. The purpose of this study was to determine the population of Ice Nucleation-Active Bacteria on Soe tangerines leaves and the class of Ice Nucleation-Active bacteria based on Ice formation temperatures. The collecting of Soe tangerine leaves used the purpose sampling method. Leaves with frost blotches were collected from three stages at altitudes of 1500, 1800, and 2000 meters above sea level (m asl). Bacterial isolation was carried out by the spread plate method on Nutrien Agar 2,5 % glycerol (NAG)  media. Ice Nucleation activity was determined by the tube nucleation test method. Estimation of INA bacterial population was conducted by the multiple-tube nucleation test with Thomas series .3.3.3. The result showed that the highest INA bacterial population was 6.9x104 which was found in leaves samples collected from stations 1800 and 2000 m asl, and the lowest population i.e. 5,4x103 on leaf samples from station 1500 m asl. Based on the temperature of ice formation, it was known that INA bacteria that attack the Soe tangerines leaves Mutis plateau are the INA bacteria class B and C.

IMPROVING ICE FORMATION BY ADDITIVE FOR COLD ENERGY STORAGE

2014 ◽  
Vol 22 (03) ◽  
pp. 1450012 ◽  
Author(s):  
JIN HU ◽  
OSMANN SARI ◽  
CYRIL MAHMED
Keyword(s):  
Heat Release ◽  
Pseudomonas Syringae ◽  
Freezing Temperature ◽  
Ice Nucleation ◽  
Ice Formation ◽  
Nucleation Agent ◽  
Cold Energy ◽  
Nucleation Activity ◽  
Effective Use ◽  
Ice Nucleation Activity

Ice storage is one technique for effective use of thermal energy. Application of bionucleant (a protein from the bacterium Pseudomonas syringae) as a snow inducer in ski field has shown great potential to enhance the quantity of snow and increase freezing temperature. In this study, differential scanning calorimeter (DSC) and lab-built ice formation reactor were employed to study experimentally the heterogeneous ice nucleation under super-cooled conditions at different dissolved bionucleant concentrations. It was found the degree of supercooling is reduced by addition of bionucleant. However, ice nucleation-activity of bionucleant will drop down when bionucleant solution is saturated/supersaturated. In our DSC measured heat release study, when bionucleant acts as ice nucleation agent in aqueous solution, prior to reaching its saturation/supersaturation, there is an increase in latent heat release during freezing/melting as the amount of dissolved bionucleant increases. In another test, the supercooling does not occur in 0.5% bionucleant solution, it began to freeze around 0°C. Our results suggest that, the addition of bionucleant may help induce ice nucleation and increase freezing temperature thereby reduces the energy consumption of ice formation for cold storage.

scholarly journals The Ice Nucleating Protein InaZ is Activated by Low Temperature

2020 ◽  
Author(s):  
Steven J. Roeters ◽  
Thaddeus W. Golbek ◽  
Mikkel Bregnhøj ◽  
Taner Drace ◽  
Sarah Alamdari ◽  
...  
Keyword(s):  
Pseudomonas Syringae ◽  
Molecular Mechanisms ◽  
Helical Structure ◽  
Frost Damage ◽  
Ice Nucleation ◽  
Structural Basis ◽  
Sum Frequency ◽  
Ina Bacteria ◽  
Dynamics Simulations ◽  
Global Precipitation

AbstractIce-nucleation active (INA) bacteria can promote the growth of ice more effectively than any other known material. Utilizing specialized ice-nucleating proteins (INPros), they obtain nutrients from plants by inducing frost damage and, when airborne in the atmosphere, they drive ice nucleation within clouds and may affect global precipitation patterns. Despite their evident environmental importance, the molecular mechanisms behind INPro-induced freezing have remained largely elusive. In the present study, we investigated the folding and the structural basis for interactions between water and the ice-nucleating protein InaZ from the INA bacterium Pseudomonas syringae strain R10.79. Using vibrational sum-frequency generation and two-dimensional infrared spectroscopy, we demonstrate that the ice-active repeats of InaZ adopt a β-helical structure in solution and at water surfaces. In this configuration, hydrogen bonding between INPros and water molecules imposes structural ordering on the adjacent water network. The observed order of water increases as the interface is cooled to temperatures close to the melting point of water. Experimental SFG data combined with spectral calculations and molecular-dynamics simulations shows that the INPro reorients at lower temperatures. We suggest that the reorientation can enhance order-inducing water interactions and, thereby, the effectiveness of ice nucleation by InaZ.

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