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PubMed2006

Rapid cold-hardening increases membrane fluidity and cold tolerance of insect cells.

The rapid cold-hardening (RCH) response not only confers dramatic protection against cold-shock (non-freezing) injury, but also "instantaneously" enhances organismal performance. Since cold-shock injury is associated with damage to the cell membrane, we investigated the relationship between RCH and changes in cold tolerance and membrane fluidity at the cellular level. None of the adult flies (Sarcophaga bullata) in the cold-shocked treatment group survived direct transfer to -8 degrees C for 2 h; in contrast, 64.5% of flies in the RCH group survived exposure to -8 degrees C. Differences between the treatment groups also were reflected at the cellular level; only 21.3% of fat body cells in the cold-shocked group survived compared to 68.5% in the RCH group. Using 31P solid-state NMR spectroscopy, we determined that membrane fluidity increased concurrently with rapid cold-hardening of fat body cells. This result suggests that membrane characteristics may be modified very rapidly to protect cells against cold-shock injury.

Adaptation, Physiological↗
PubMed1987

Cold tolerance in mammalian cells.

As whole organisms, most mammals have a poor tolerance for hypothermia. But their cells may have a capacity for a far wider cold tolerance, which may be expressed in peripheral tissues, sporadically in core tissue and in cultured cells. Against this background the cold resistance of cells of deep hibernators may be seen as the extreme of a continuum and is complicated by the consideration that the voluntary hypothermia of hibernation is probably in most cases a metabolic adaptation to forestall starvation. Similarly, cold resistance of peripheral tissues may in diving animals be confounded by the need to be adapted to hypoxia as well. Hence, attempts to analyse cold resistance by comparisons of absolute rates of arbitrarily chosen reactions may be misleading. A more useful approach is analysis of maintenance of balance: balance between ATP synthesis and utilization, balance between macromolecule synthesis and degradation and balance between pumps and leaks. Cation pumps and leaks constitute a major component of energy utilization and are central to other cell functions, even during minimal metabolism. Hence, the maintenance of ion gradients is a central issue in understanding adaptation not only to hypothermia but also to starvation and hypothermia. Of the three hypometabolic states, hypothermia has been best studied in this regard. In most cases, passive permeability is more reduced at low temperature in cold-tolerant cells than in cold-sensitive ones. In some cases there is also a difference in Na-K pump activity and perhaps in ATP dependent Ca-pump activity. Pump activities and probably the maintenance of minimal leak require ongoing metabolism. The question of whether, in cold-sensitive cells, energy supplies are adequate at low temperature was once the focus of this field, but has been ignored for a decade without having been fully resolved. There are many instances of less temperature sensitivity of specific metabolic activities (mitochondrial respiration, etc.) in hibernators than in non-hibernators, without any verification of whether this is essential for survival at low body temperature. Certainly, robust pumping has been found in some failing cold-sensitive cells at low temperature, suggesting no shortage of ATP in these cases, but in other cases the issue may be a more complex one than just that of ATP availability.(ABSTRACT TRUNCATED AT 400 WORDS)

Adaptation, Physiological↗
PubMed1973

Short term increases in the cold tolerance of red osier dogwood stems induced by application of cysteine.

Bark tissues of Cornus stolonifera stems, treated with cysteine at 24 hours after treatment, survived exposure to -11 C (the tissue temperature) with little or no injury. An initiation of increase in the cold tolerance was usually observed when plants were treated with cysteine at 12 hours after treatment. Neither plants at 36 or 48 hours after treatment nor plants 12 hours before treatment had shown increases in the cold tolerance. They were killed below -5 C, which was the survival temperature of untreated control plants. Two weeks or more of short day induction before cysteine application were required for a significant effect of short term 5 C increase in the cold tolerance.

Journal Article↗
PubMed1992

Effects of radiational heating at low air temperature on water balance, cold tolerance, and visible injury of red spruce foliage.

Recent studies have shown that winter needle mortality in red spruce (Picea rubens Sarg.) is increased by exposure to direct solar radiation, possibly as a result of photo-oxidative damage, accelerated winter desiccation, or reduced cold tolerance due to heating of sun-exposed needles. In an experiment at controlled subfreezing air temperatures of -10 to -20 degrees C, visible radiation was less effective than infrared radiation in producing needle desiccation and visible injury during freeze-thaw cycles. However, visible radiation produced a red-brown color in injured needles, similar to natural winter injury, whereas injured needles exposed to infrared radiation were yellow and injured needles kept in darkness were dark brown. Thus, visible radiation was necessary to produce the red-brown color of damaged needles, but not the injury itself. Needle desiccation was not strongly correlated with visible injury, but the pattern of variation in visible injury among trees and the positive correlation between electrolyte leakage and visible injury suggested that freezing damage following freeze-thaw cycles might cause the visible injury. This was confirmed by a second experiment that showed loss of cold hardiness in needles thawed by radiational heating for six consecutive days. Even with a constant nighttime temperature of -10 degrees C, six days of radiational heating of needles to above freezing caused a small (2.8 degrees C) mean decrease in needle cold tolerance, as measured by electrolyte leakage. Continuous darkness at -10 degrees C for six days resulted in an estimated 5.6 degrees C mean increase in needle cold tolerance. Freezing injury stimulated desiccation: cooling at 4 degrees C h(-1) to -43 or -48 degrees C increased the dehydration rate of isolated shoots by a factor of two to three during the first day after thawing. Within three days at 15 to 22 degrees C and 50% relative humidity, the mean water content of these shoots fell to 60% or lower, compared to 90% or greater for unfrozen controls or shoots subject to less severe freezing stress. In some but not all severely freeze-stressed shoots, accelerated needle desiccation and abscission were accompanied by a red-brown color typical of red spruce winter needle injury. We conclude that severe winter desiccation in red spruce may often be due to prior freezing injury, increased as a result of exposure to direct solar radiation. Furthermore, freezing injury in red spruce may sometimes cause desiccation and abscission of green needles.

Journal Article↗
PubMed2006

[Cold tolerance and wintering cultivation effect of different Welsh onion varieties].

With Welsh onion Zhangqiu as the contrast, this paper measured the physiological indices including electrolyte leakage, malondialdehyde content, protective enzyme activity, chlorophyll content, photosynthetic rate and root vitality of two introduced Welsh onion varieties Chunwei and Changhao, and identified their cold tolerance and wintering cultivation effect. The results showed that during wintering cultivation, all test Welsh onion varieties suffered low temperature stress, which led the electrolyte leakage and malondialdehyde content arrived at the maximum, while the chlorophyll content and photosynthetic rate dropped to the bottom at 15 January. At this time, the function data of physiological indices were 0. 452, 0. 364, and 0. 226 for Chuowei, Changbao and Zhangqiu, respectively, suggesting that Chunwei had stronger cold tolerance, followed by Changbao, and Zhangqiu. The bolting rate of Chunwei, Changbao and Zhangqiu when harvested was 0, 35.2% and 81.0% , respectively. Although the biological yield of Changbao was 25.67% and 52. 94% higher than that of Chunwei and Zhangqiu, respectively, the economic yield of Chunwei was the highest (5.49 kg m2) , with an increment of 18. 57% than Changbao, and that of Zhangqiu was only 0. 86 kg x m(-2) It could be concluded that Chonwei was fit to cultivate in low tunnel in winter, while Zhangqiu was weaker in cold tolerance and not fit to wintering cultivation.

Allium↗
PubMed1975

Differential cold tolerance, starch, sugar, protein, and lipid of yellow and purple nutsedge tubers.

From measurements of viability after exposure of tubers to natural overwintering in the soil and 6 weeks exposure at 2 C, species cold tolerance of the tubers was ranked in decreasing order: yellow nutsedge ;I' (Cyperus esculentus L.), an ecotype originating in Illinois; yellow nutsedge ;G', an ecotype originating in Georgia; and purple nutsedge (Cyperus rotundus L.). The ratios of unsaturated-saturated fatty acids in tuber triglycerides, tuber polar lipids, and leaf polar lipids followed the same order as the cold tolerance rankings, with the most cold-hardy species having the highest ratios. Lipid content was less than 1% of dry weight in purple nutsedge tubers, but was from 5 to 7% in both yellow nutsedge tubers. Starch, sugar, and lipid contents increased significantly in the hardy yellow nutsedge ;I' tubers during a 6-week exposure to 2 C, but did not change in the susceptible purple nutsedge tubers; only sugar increased in yellow nutsedge ;G' tubers after this treatment. Protein content was not altered by the 2 C treatment in any of the tubers. Apparently, several factors involving starch, sugar, lipids, and fatty acids are related to the differences in tolerance to cold in these species.

Journal Article↗
PubMed2004

Biodegradation of phenol and phenol-related compounds by psychrophilic and cold-tolerant alpine yeasts.

We characterized 32 cold-adapted, psychrophilic and cold-tolerant, yeast strains isolated from alpine habitats with regard to their taxonomy, growth temperature profile, and ability to degrade phenol and 18 phenol-related mono-aromatic compounds at 10 degrees C. Twenty of the strains were identified by sequencing of the ribosomal ITS region as seven species of the basidiomycota: Cryptococcus terreus (three strains), Cryptococcus terricola (one strain), Rhodosporidium lusitaniae (two strains), Rhodotorula creatinivora (10 strains), Rhodotorula ingeniosa (one strain), Mastigobasidium intermedium (one strain), and Sporobolomyces roseus (two strains). Twelve strains sharing closely related ITS sequences could not be identified to the species level; according to their ITS sequence they are included in the Microbotryomycetidae. These 12 strains were psychrophilic (no growth at temperatures above 20 degrees C); one-third of these strains did not grow above 15 degrees C. None of the 32 strains utilized any of the highly volatile mono-aromatic compounds (benzene, toluene, ethylbenzene, nitrobenzene, o-xylene, m-xylene, and p-xylene) as the sole carbon source. Non/low volatile aromatic compounds were degraded in the following order: phenol>hydroquinone>resorcinol>benzoate>catechol>salicylate>>p-cresol>m-cresol. o-Cresol, guaiacol, p-nitrophenol, or p-nitrotoluene were not utilized for growth. R. creatinivora strains degraded up to seven compounds, whereas C. terricola and S. roseus strains degraded only two compounds. The toxicity of the compounds was determined via growth inhibition in the presence of toxicants and nutrients at 10 degrees C. R. creatinivora strains were characterized by higher IC50 values than other species, S. roseus was the most sensitive species. The most toxic compounds were the xylene isomers, ethylbenzene, p-nitrophenol, and m-cresol. There was a relation between the chemical structure of the compounds and their toxicity, whereas a relation between the toxicity of the compounds and the ability of the yeasts strains to utilize these compounds for growth was only detected in some cases.

Adaptation, Physiological↗
PubMed1989

Mechanism of enhanced cold tolerance by an ephedrine-caffeine mixture in humans.

The influence of a thermogenic mixture of ephedrine- (1 mg/kg) caffeine (2.5 mg/kg) on cold tolerance was investigated in nine healthy young male subjects during two seminude exposures to cold air (3 h at 10 degrees C). The drug ingestion reduced the total drop in core, mean skin, and mean body temperatures (P less than 0.01), thus producing significantly warmer final core, mean skin, and mean body temperatures compared with the placebo ingestion. The drug ingestion increased the total 3-h energy expenditure by 18.6% compared with that of the placebo ingestion in the cold (P less than 0.01). By means of the nonprotein respiratory exchange ratio to calculate the rates of substrate oxidation, it was found that the drug ingestion increased carbohydrate oxidation by as much as 41.7% above that of the placebo (P less than 0.05). In contrast, the drug mixture had no significant influence on lipid or protein metabolism. The results demonstrate that the ingestion of an ephedrine-caffeine mixture improves cold tolerance in humans by significantly increasing body temperatures in the cold. These improvements were not caused by an increased conservation of heat but by a greater energy expenditure, which appears to be dependent on an enhanced carbohydrate utilization.

Adult↗
PubMed1990

Improvement of cold tolerance by selective A1 adenosine receptor antagonists in rats.

Previously we have shown that the improvement of cold tolerance by theophylline is due to antagonism at adenosine receptors rather than inhibition of phosphodiesterase. Since theophylline is a nonselective adenosine receptor antagonist for both A1 and A2 receptors, the present study investigated the adenosine receptor subtype involved in theophylline's action. Acute systemic injection of selective A1 receptor antagonists (1,3-dialkyl-8-aryl or 1,3-dialkyl-8-cyclopentyl xanthine derivatives) significantly increased both the total and maximal heat production as well as cold tolerance. In contrast, injection of a relatively selective A2 receptor antagonist, 3,7-dimethyl-1-propargylxanthine (compound No. 19), failed to significantly alter the thermogenic response of the rat under cold exposure. Further, the relative effectiveness of these compounds in increasing total thermogenesis was positively correlated with their potency in blocking the A1 adenosine receptor (r = .52, p less than 0.01), but not in A2 adenosine receptor (r = .20, p less than 0.2). It is likely that the thermally beneficial effects of adenosine A1 antagonists are due to their attenuation of the inhibitory effects of endogenously released adenosine on lipolysis and glucose utilization, resulting in increased substrate mobilization and utilization for enhanced thermogenesis.

Animals↗
PubMed2003

Mapping of QTLs associated with cold tolerance during the vegetative stage in rice.

Low-temperature stress is an important factor affecting the growth and development of rice (Oryza sativa L.) in temperate and high-elevation areas. Cold stress may cause various seedling injuries, delayed heading and yield reduction due to spikelet sterility. In this study, 181 microsatellite marker loci were used to identify quantitative trait loci (QTLs) associated with cold tolerance at the vegetative stage in 191 recombinant inbred lines (RILs) derived from a cross of a cold-tolerant temperate japonica cultivar (M-202) with a cold-sensitive indica cultivar (IR50). Different temperature regimes were applied in growth chambers on 191 RILs. The temperature regimes imposed in the growth chamber simulated cold-stress injuries at the seedling and late vegetative stages. In this study a major QTL was identified on chromosome 12, designated as qCTS12a, that was closely associated with cold-induced necrosis and wilting tolerance, and accounted for 41% of the phenotypic variation. A number of QTLs with smaller effects were also detected on eight rice chromosomes.

Acclimatization↗
PubMed1976

Extractant Influence on the Relationship between Extractable Proteins and Cold Tolerance of Alfalfa.

The influence of ionic composition and pH of extractant on the relationship between the extracted proteins and the cold tolerance of Vernal and Arizona Common alfalfa (Medicago sativa L.) was examined. Five environments were used to induce different tolerance levels. The quantity of protein extracted from plants was influenced by the hardening environment, cultivar, and ionic composition and pH of 29 extractants. Extractants with a pH below 6 generally extracted less protein.The measured cold tolerance of the plants was correlated with the quantity of protein detected in many of the 14 regions of the electrophoresis gel columns regardless of extractant but was most closely associated with the protein in either region 7 or 8 with nine of ten extractants.The magnitudes of cultivar and hardening effects on quantities of protein detected on various column regions were influenced by choice of extractant. This suggests that proper extractant selection may be vital for study of isozymes by electrophoresis.

Journal Article↗
PubMed1990

Do adenosine antagonists improve cold tolerance by reducing hypothalamic adenosine activity in rats?

Previously we have shown that systemic injection of adenosine antagonists can significantly improve cold tolerance in both rats and humans. However, it is not clear whether systemic administration of adenosine antagonist acts peripherally or centrally at the thermoregulatory site. To resolve this, theophylline (nonselective adenosine receptor blocker), cyclopentyltheophylline (selective A1 receptor blocker) or adenosine deaminase (an enzyme which inactivates adenosine by converting it into inosine) was injected directly into preoptic anterior hypothalamus (POAH) of rats and their thermogenic responses assessed. In contrast to that observed after systemic administration, intrahypothalamic injection of either adenosine antagonists or deaminase at various doses failed to elicit any enhancement in heat production beyond that of the controls. These results suggest that the beneficial effect of systemically injected adenosine antagonists in improving cold tolerance is not the result of altering the thermoregulatory functions mediated via the POAH.

Adenosine Deaminase↗
PubMed1991

Cold tolerance and metabolic heat production in male C57BL/6J mice at different times of day.

Nine-month-old male C57BL/6J mice were subjected to three-hour cold stress tests (partial restraint at 6 degrees C) at 9:00 a.m. or at 1:00 p.m. Tests were repeated three times at two-week intervals at the same time of day. Body temperature was measured by colonic thermoprobe, and metabolic heat production was measured by indirect calorimetry during each test. All mice showed habituation to repeated cold exposures (an improvement of cold tolerance across tests) due to an increase in metabolic heat production. The levels of metabolic heat production were similar during morning and afternoon testing; however, mice tested in the afternoon had consistently poorer cold tolerance, which indicated increased heat loss. Increased heat loss in mice of similar body weight and presumably similar body composition, suggests that there is less effective cold-induced skin vasoconstriction during the afternoon. We hypothesize that the compromised skin vasomotor response during the afternoon cold exposure results from competing effects of vasodilation due to local autoregulation stimulated by a circadian reduction of cardiac output during the sleep phase, and vasoconstriction due to the cold stress.

Animals↗
PubMed1998

3,5-Diiodo-L-thyronine and 3,5,3'-triiodo-L-thyronine both improve the cold tolerance of hypothyroid rats, but possibly via different mechanisms.

The effects of 3,5-diiodo-L-thyronine (3,5-T2, 2.5-10 microg/100 g BW) on cold tolerance, energy expenditure and oxidative capacity of four metabolically very active tissues (brown adipose tissue, skeletal muscle, liver and heart) were determined in hypothyroid, cold-exposed rats. Hypothyroid rats survived cold for only 3-4 days. 3,5-T2 improved survival dose dependently; with 10 microg/100 g BW the rats survived 3 weeks (limit of observation). This effect was paralleled by an increased energy expenditure of the whole animal for the entire 3 weeks. Similar effects were observed in hypothyroid rats treated with 3,3',5-triiodo-L-thyronine (T3). 3,5-T2 stimulated the specific oxidative capacity (expressed as cytochrome oxidase activity per milligram protein) of all four tissues dose dependently. When the oxidative capacity was expressed as total activity (cytochrome oxidase activity times organ weight), the percentage increases were of the same order. T3 exerted similar effects, but the changes in total activity were much greater than in specific activity, indicating an effect on the tissue trophism. The effect of 3,5-T2 on cold tolerance thus mimics the effect of T3, but via different cellular mechanisms. T3 seems to act primarily on the trophism of the tissues, while 3,5-T2 may act directly on mitochondria without an effect on tissue trophism.

Adaptation, Physiological↗
PubMed2006

Responses of Picea mariana to elevated CO2 concentration during growth, cold hardening and dehardening: phenology, cold tolerance, photosynthesis and growth.

Seedlings from a northern and a southern provenance of black spruce (Picea mariana Mill. BSP) from eastern Canada were exposed to 37 or 71 Pa of carbon dioxide (CO2) during growth, cold hardening and dehardening in a greenhouse. Bud phenology, cold tolerance and photosynthetic efficiency were assessed during the growing and over-wintering periods. Bud set occurred earlier in elevated [CO2] than in ambient [CO2], but it was later in the southern provenance than in the northern provenance. An increase in seedling cold tolerance in early fall was related to early bud set in elevated [CO2]. Maximal photosystem II (PSII) photochemical efficiency (F(v)/F(m)), effective quantum yield (phi(PSII)), photochemical quenching (q(P)), light-saturated photosynthesis (Amax), apparent quantum efficiency (alpha'), light-saturated rate of carboxylation (Vcmax) and electron transport (Jmax) decreased during hardening and recovered during dehardening. Although Amax and alpha' were higher in elevated [CO2] when measured at the growth [CO2], down-regulation of photosynthesis occurred in elevated [CO2] as shown by lower F(v)/F(m), phi(PSII), Vcmax and Jmax. Elevated [CO2] reduced gene expression of the small subunit of Rubisco and also decreased chlorophyll a/chlorophyll b ratio and nitrogen concentration in needles, confirming our observation of down-regulation of photosynthesis. Elevated [CO2] increased the CO2 diffusion gradient and decreased photorespiration, which may have contributed to enhance Amax despite down-regulation of photosynthesis. Total seedling dry mass was higher in elevated [CO2] than in ambient [CO2] at the end of the growing season. However, because of earlier bud formation and cold hardening, and down-regulation of photosynthesis during fall and winter in elevated [CO2], the treatment difference in dry mass increment was less by the end of the winter than during the growing season. Differences in photosynthetic rate observed during fall, winter and spring account for the inter-annual variations in carbon assimilation of black spruce seedlings: our results demonstrate that these variations need to be considered in carbon budget studies.

Carbohydrate Metabolism↗
PubMed2004

Expression of cold-tolerant pyruvate, orthophosphate dikinase cDNA, and heterotetramer formation in transgenic maize plants.

Maize is a typical C4 plant of the NADP-malic enzyme type, and its high productivity is supported by the C4 photosynthetic cycle, which concentrates atmospheric CO2 in the leaves. The plant exhibits superior photosynthetic ability under high light and high temperature, but under cold conditions the photosynthetic rate is significantly reduced. Pyruvate orthophosphate dikinase (PPDK), a key enzyme of the C4 pathway in maize, loses its activity below about 12 degrees C by dissociation of the tetramer and it is considered as one possible cause of the reduction in the photosynthetic rate of maize at low temperatures. To improve the cold stability of the enzyme, we introduced a cold-tolerant PPDK cDNA isolated from Flaveria brownii into maize by Agrobacterium-mediated transformation. We obtained higher levels of expression by using a double intron cassette and a chimeric cDNA made from F. bidentis and F. brownii with a maximum content of I mg/g fresh weight. In leaves of transgenic maize, PPDK molecules produced from the transgene were detected in cold-tolerant homotetramers or in heterotetramers of intermediate cold susceptibility formed with the internal PPDK. Simultaneous introduction of an antisense gene for maize PPDK generated plants in which the ratio of heterolologous and endogenous PPDK was greatly improved. Arrhenius plot analysis of the enzyme extracted from one such plant revealed that the break point was shifted about 3 degrees C lower than that of the wild type.

Cold Temperature↗
PubMed1992

Topical application of ice-nucleating-active bacteria decreases insect cold tolerance.

The majority of overwintering insects avoid lethal freezing by lowering the temperature at which ice spontaneously nucleates within their body fluids. We examined the effect of ice-nucleating-active bacteria on the cold-hardiness of the lady beetle, Hippodamia convergens, a freeze-intolerant species that overwinters by supercooling to ca. -16 degrees C. Topical application of the ice-nucleating-active bacteria Pseudomonas syringae increased the supercooling point to temperatures as high as -3 degrees C. This decrease in cold tolerance was maintained for at least 3 days after treatment. Various treatment doses (10, 10, and 10 bacteria per ml) and modes of action (bacterial ingestion and topical application) were also compared. At the highest concentration of topically applied P. syringae, 50% of the beetles froze between -2 and -4 degrees C. After topical application at the lowest concentration, 50% of the individuals froze by -11 degrees C. In contrast, beetles fed bacteria at this concentration did not begin to freeze until -10 degrees C, and 50% were frozen only at temperatures of -13 degrees C or less. In addition to reducing the supercooling capacity in H. convergens, ice-nucleating-active bacteria also significantly reduced the cold-hardiness of four additional insects. These data demonstrate that ice-nucleating-active bacteria can be used to elevate the supercooling point and thereby decrease insect cold tolerance. The results of this study support the proposition that ice-nucleating-active bacteria may be used as a biological insecticide for the control of insect pests during the winter.

Journal Article↗
PubMed2026

DNA hypermethylation of abscisic-acid-related genes helps enhance the cold tolerance of tetraploid rice.

Polyploid plants exhibit enhanced stress resistance and superior adaptability to extreme environments, but the underlying molecular mechanisms remain incompletely understood. Here we confirm that tetraploid rice exhibits stronger cold tolerance than diploid rice. This improved tolerance is mediated by reduced malondialdehyde accumulation, elevated antioxidant enzyme activity, and epigenetic regulation of genes involved in abscisic acid (ABA) biosynthesis and signaling. Under cold stress, tetraploid rice induces stress-responsive genes (especially in the ABA pathway) more rapidly and to higher levels than diploid rice. This enhanced gene expression coincides with increased endogenous ABA accumulation. Furthermore, polyploidization and cold stress synergistically induce high methylation at CG, CHG, and CHH sites in genes and transposons (TEs). Notably, the methylation level of class II TEs in tetraploid rice is significantly higher than in diploid rice under low temperatures. To suppress TE activation in gene promoter regions under cold stress, tetraploid rice enhances the methylation level of ABA pathway-related gene promoters, thereby silencing TEs and maintaining genome stability. Collectively, these results enrich the theoretical understanding of the strong stress tolerance in polyploid plants and provide theoretical support for breeding cold-tolerant polyploid rice varieties.

ABA↗