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Genome-Wide Identification of SSR and InDel Markers and Experimental Validation of SSR Markers for Distinguishing Cold-Tolerant and Cold-Sensitive Lily Cultivars.

In this study, whole-genome resequencing was performed on the cold-tolerant variety ND-6 and the cold-sensitive variety 'Sorbonne'. After evaluation, the Lilium davidii var. unicolor reference genome was selected to analyze SSR distribution characteristics. Whole-genome InDel identification and comparative analysis were conducted for the two varieties, yielding 34,812,909 and 24,497,857 InDels, respectively. Short InDels were predominant, with deletions slightly outnumbering insertions, mostly located in intergenic regions. Twenty pairs of SSR primers were screened and synthesized. Among them, 10 pairs amplified clearly, with a polymorphism rate of 82.6%, effectively distinguishing the two cultivars examined in this study. This study provides systematic data and a reliable marker resource for the analysis of lily genomic variation, laying a foundation for the identification of cold-tolerant germplasm; validation across additional cultivars and individuals will be required to extend their utility to broader germplasm.

cold resistant lilies

[Positive cross-adaptation between endurance physical training and general cold tolerance to acute cold exposure in rats].

This investigation suggested that a series of endurance physical training in rats could cause an improved cold tolerance in rats. The warm-acclimated control group was kept in a thermally neutral environment (25 +/- 1 degrees C). The trained group was subjected to forced physical training on a treadmill, exercising once 1 hour daily, and five times per week for the periods of 3 to 13 weeks. The cold-acclimated group was exposed to an ambient temperature of 5.0 +/- 0.5 degrees C for 12 weeks. Improved endurance physical fitness was suggested by a cardiac hypertrophy. The positive correlation was shown between the amount or intensity of training and the enhanced thermogenesis to acute cold exposure. However, no effect of endurance training was observed on the metabolic responses to noradrenaline and adrenaline. It was inferred that the acute cold exposure caused greater utilization of blood glucose in the trained rats than in the warm controls, but did not FFA in the latter. The increases in oxygen consumption and colonic temperature were caused by hexamethonium injection during cold exposure in the trained rats, but not in the warm and cold-acclimated rats. The increased metabolic rate to acute cold exposure was improved by the prolonged physical training. The development of greater metabolic rate during acute cold exposure in the trained rats was indicated to be associated with increased noradrenaline and adrenaline-independent non-shivering thermogenesises as well as enhanced resting metabolic rate at 25 degrees C ambient temperature.

Adaptation, Physiological

Effects of diets on cold tolerance and metabolic responses to cold in fasted rats.

Effects of high-fat and high-protein diets on cold tolerance in fasted rats were investigated. High-fat diets caused significant increases in body weight, blood-free fatty acids (FFA), ketone bodies and glucose, while high-protein diet did not modify any of these parameters. Rats on high-fat diets that were exposed to cold after clipping exhibited an intermediate cold tolerance as assessed by the rate of fall in colonic temperature between control rats on a standard diet and cold-acclimated rats. The extent of increase of blood FFA and decrease of blood glucose due to cold exposure was less in the high-fat diet group than in control group, but greater than in cold-acclimated group. The lower fall in colonic temperature due to cold exposure was signifcantly associated with less increase in blood FFA and less decrease in blood glucose. In this relation the high-fat diet group was also intermediate between the control and cold-acclimated groups. The high-protein diet did not make any difference in cold tolerance and cold-induced changes in blood metabolites as compared with those in control standard diet, although it resulted in a marked increase in urinary nitrogen excretion. These results indicate that a high-fat diet could exert a significant favorable effect on cold tolerance in fasted rats, but the effect would not be as much as as in cold acclimated rats.

Adaptation, Physiological

Improved cold tolerance and its mechanism in cold-acclimated rats by high fat diet feeding.

Cold tolerance and metabolic responses to cold were studied in cold-acclimated rats on high fat diet (CAHF). Cold tolerance at-5 degrees C was assessed by fall of colonic temperature of clipped rats after 18 h of fasting. Rate of fall in colonic temperature was greatest in warm-acclimated control rats (WAST), slowest in cold-acclimated rats on standard diet (CAST), and remained unchanged in CAHF during cold exposure for 240 min. Increment in blood free fatty acid (FFA) concentration 80 min after cold exposure was greatest in WAST, less in CAST, and least in CAHF. Blood glucose decreased similarly in WAST and CAST after cold exposure, while it remained unchanged in CAHF. Blood beta-hydroxybutyrate also increased similarly in WAST and CAST, while it did not change in CAHF. Nonshivering thermogenesis tested by noradrenaline was greatest in CAHF, followed by CAST and WAST. Shivering induced by cold exposure was less pronounced in CAST than in WAST and did not develop in CAHF; changes in colonic temperature were inversely related to the extent of shivering during cold exposure for 90 min. These results suggest that an integrating effect of cold and high fat diet could improve cold tolerance much more than cold acclimation itself, possibly through enhanced nonshivering thermogenesis caused by metabolic modifications such as increased lipid use and gluconeogenesis.

Acclimatization

Lowered cold tolerance in cold-acclimated and non-acclimated guinea pigs treated with diazepam.

The effects of the clinically most commonly used minor tranquilizer, diazepam, on the survival time and on the mechanism of death in non-acclimated and cold-acclimated guinea pigs in severe cold exposure (-20 degrees C) were studied. Cold acclimation for 2 months increased the average survival time from 4 h to 10 h. The lowest rectal temperature at death (14.6 degrees C) was seen in the cold-acclimated animals. Diazepam at a dose of 5 or 15 mg/kg i.p. 30 min before the beginning of the exposure reduced dose-dependently the cold endurance of both cold-acclimated and non-acclimated guinea pigs. The serum glucose and free fatty acid concentrations were low in the animals with the long survival time. Histological studies of liver, kidney, and adrenal glands showed no specific changes. Exposure seemed to increase the frequency of contraction bands and to decrease focally the intensity of beta-hydroxybutyrate dehydrogenase reaction in the myocardium, which indicates a mild hypoxic lesion of the muscle cells.

Acclimatization

Integrated 16 S rRNA and transcriptome analysis reveal molecular and microbial mechanisms of cold-tolerant germination in hulless barley.

BACKGROUND: Elucidating the mechanisms underlying cold-tolerant germination is crucial for enhancing crop resilience to low temperatures. Hulless barley (Hordeum vulgare var. coeleste L.), with remarkable natural cold adaptation, serves as an ideal model to study cold stress tolerance mechanisms in gramineous crops. In this study, cold-tolerant variety 37 and cold-sensitive variety 44 were screened and used to investigate the molecular mechanisms of cold-tolerant germination, via seed germination assays, combined with phytohormone determination, transcriptome sequencing and 16 S rRNA amplicon sequencing. RESULTS: Low temperature significantly inhibited hulless barley seed germination: the germination rate of cold-sensitive variety 44 decreased by 69%, while that of cold-tolerant variety 37 only decreased by 2%. Transcriptome analysis identified 2,647 and 2,392 differentially expressed genes (DEGs) in variety 37 and 44, respectively. Weighted gene co-expression network analysis (WGCNA) revealed a green module significantly positively correlated with gibberellic acid (GA) content, containing 10 core genes such as late embryogenesis abundant protein (LEA) and Homeobox genes. 16 S rRNA sequencing showed that the cold-tolerant variety 37 had enriched abundances of dominant endophytes including Sphingomonas and Pelomonas, with correlation coefficients of 0.70 and 0.87 with GA content, respectively. Additionally, exogenous GA treatment significantly increased germination rates under cold stress by 176.67% in cold-sensitive variety 44. CONCLUSIONS: This study confirms that the enhanced cold tolerance of hulless barley during seed germination originates from the synergistic interaction between beneficial endophytes (Sphingomonas, Pelomonas), GA, and core genes (e.g., LEA, Homeobox). Exogenous GA application can significantly restore the germination ability of cold-sensitive varieties. These findings provide a critical theoretical basis for improving cold tolerance in hulless barley germplasm.

Hordeum

Improving cold tolerance in elderly rats by aminophylline.

During severe cold exposure, old rats (23-26 months) were less capable in maintaining normal body temperature as compared to young rats (6-9 months) due to lower rate of heat production (HP). Single injection of optimal doses of aminophylline (AMPY; 10 and 18.7 mg/kg, i.p.), a phosphodiesterase inhibitor which enhances the intracellular cyclic AMP concentration, significantly increased the rate of HP in old rats to levels beyond the control values observed in young rats. Consequently, cold tolerance of the old rats was significantly improved. This AMPY-improved cold tolerance is apparently not due to increased non-shivering thermogenesis (NST) since AMPY failed to enhance norepinephrine-stimulated NST in the old rats. It is likely that AMPY increased substrate mobilization and/or conversion, thereby circumventing the limiting role of substrate availability for shivering thermogenesis. Thus, the age-dependent decrease in cold tolerance may be due to a reduced capacity for substrate mobilization when challenged by cold.

Aging

Effect of aging on cold tolerance and thyroid activity in CBA/Ca inbred mice.

Relationships between cold tolerance, serum levels of thyroxine (T4), thyronine (T3), and thyrotropine (TSH), and thyroid morphometry have been investigated in male CBA/Ca inbred mice at various ages through their life span. From the data obtained it appeared that there was an age-related decrease in cold tolerance up to 18 months of age which was followed by an increase, the age effect being most apparent in relation to cold resistance and cold tolerance during the recovery period following cold exposure. The age-related changes in cold tolerance appeared to be associated with changes in the serum concentrations of T3, T4, and TSH. In contrast to the T3 serum levels which showed a decrease at 36 months, the thyroxine contents showed a perceptible decrease from the age of 12-18 months onwards. A similar pattern was observed for the TSH levels, with a peak at 21 months, followed by a decline at 30 months. A relationship with age between serum T4 level and thyroid weight was indicated together with structural changes in the thyroid gland, particularly during senescence, for example the size and number of thyroid epithelial cells had become enlarged by 30 months of age.

Acclimatization

The effect of cold acclimation and exercise training on cold tolerance in aged C57BL/6J mice.

We investigated whether intermittent cold acclimation and low intensity exercise training, two interventions known to improve thermoregulatory function in young rodents, enhanced cold tolerance in aged C57BL/6J mice. Mice, aged 18 to 20 months, were randomly assigned to four treatment conditions: (a) intermittent cold acclimation (CA) (50 min per day, 5 times per week for 3 weeks at 5 degrees C), (b) submaximal treadmill exercise (EX) (15 m min-1 for 30 min, 5 times per week for 8 weeks, (c) sequential treatment of CA + EX, and (d) control group. Mice were exposed to a 3-hr cold stress test (15 degrees C) prior to and following treatment. CA aged mice maintained normothermia and demonstrated significant increases in oxygen consumption and brown adipose tissue protein concentration compared with controls; enhanced cold tolerance was probably due to increased utilization of nonshivering thermogenesis. EX aged mice had elevated O2 consumption and increased skeletal muscle enzyme activity compared with controls; however, cold tolerance was not enhanced compared with CA mice. The data suggest that intermittent CA effectively reduces hypothermia during cold challenge in aged mice.

Acclimatization

Age comparisons of body temperature and cold tolerance among different strains of Mus musculus.

The age-related declines in colonic temperature (Tco) and cold tolerance (ability to maintain Tco when exposed to 10 degrees C for 3 h) described for C57BL/6J mice are compared to other mouse strains. Assessment of young and aged male mice of the C57BL/6J and A/J inbred strains and their F1 hybrid, B6AF1/J, as well as a pen-bred strain of Mus musculus captured from the wild revealed an aged-related decline in thermoregulation among all these strains. The degree of decline in thermoregulation was roughly correlated to differences in strain-specific lifespan. Aged mice of the relatively short-lived genotype, A/J (mean lifespan of 22 months), had the lowest Tco and poorest cold tolerance. The long-lived hybrids, B6AF1/J (mean lifespan of 29 months), demonstrated the highest Tco and the best cold tolerance among aged mice. C57BL/6J (mean lifespan of 26 months) showed an intermediate level of thermoregulation. Aged pen-bred mice demonstrated a significant decline in cold tolerance, but not a significant decline in Tco. The thermoregulatory responses of the pen-bred mice were superior to those observed among the domesticated strains. These data suggest that the age-related impairment in thermoregulation is a general phenomenon of aging in Mus musculus that is correlated with strain-specific lifespan but is possibly affected by the level of hybridization and domestication.

Aging

ATP dependence of Na(+)-K+ pump of cold-sensitive and cold-tolerant mammalian red blood cells.

1. The ATP concentration of intact, cold-tolerant (ground squirrel) red cells and cold-sensitive (guinea-pig and human) red cells was monitored by use of the firefly tail, luciferin-luciferase assay. ATP kinetics of the pump in intact red blood cells was investigated by altering cell [ATP] by progressive depletion of ATP in the presence of 2-deoxy-D-glucose and then by measurement of ouabain-sensitive K+ influx at each level of [ATP] at various temperatures between 37 and 5 degrees C. Na(+)-K(+)-ATPase activity of broken membranes was also determined in parallel experiments using ouabain-sensitive release of 32P from [gamma-32P]ATP as a measure of activity. 2. Without depletion, there is no immediate decrease in [ATP] of intact cold-sensitive cells at low temperature (5 degrees C) at times when there are marked differences in the activities of the Na(+)-K+ pump of cold-tolerant and cold-sensitive cells. 3. At 37 degrees C Na(+)-K(+)-ATPase of all three species exhibited two components of ATP dependence at 37 degrees C, one with high velocity, low affinity, the other with low velocity, high affinity. Affinities of both components rose with cooling. 4. A similar, two component pattern was observed in intact guinea-pig and human red cells at 37 degrees C, except that the segment corresponding to the high affinity component had an apparent Km (Michaelis-Menten constant) 3- to 4-fold higher than that of the broken membrane preparation. 5. Cooling intact guinea-pig and human red cells decreased the apparent affinity of the high velocity, low affinity component for ATP, so that at 20 degrees C the value of Km approached or exceeded the levels of physiological ATP concentration. Below 20 degrees C only one component with values corresponding to that of the low velocity, high affinity component could be observed. 6. In intact ground squirrel cells only the low affinity, high velocity component was apparent between 37 and 5 degrees C. Its affinity for ATP rose with cooling between 37 and 5 degrees C.

Adenosine Triphosphate

Effect of reproductive function on cold tolerance in deer mice.

Thermoregulatory responses were evaluated in male deer mice (Peromyscus maniculatus nebrascensis) after exposure to short photoperiod and either warm or cold ambient temperature (T(a)). Deer mice were chosen for this study because males exhibit differential reproductive responses to short day length (SD); this difference has a genetic basis, and both phenotypes are found within natural breeding populations. Deer mice undergoing SD-induced gonadal regression significantly improved their cold limit to -32.9 degrees C after exposure to SD/warm T(a) and to -47.4 degrees C after SD/cold T(a) exposure, relative to long day length/warm T(a) controls (-17.4 degrees C). In contrast, deer mice maintaining reproductive function despite SD exposure significantly improved cold limit to -27.2 degrees C only after exposure to SD/cold T(a), relative to controls (-16.3 degrees C). Maximum norepinephrine-induced nonshivering thermogenesis (NST) did not vary with reproductive state, indicating differences in cold tolerance were not due to capacity to produce heat by NST. Comparison between phenotypes of heat production during cold tolerance tests indicated that greater cold tolerance among mice exhibiting SD-induced gonadal regression can be accounted for by 1) lower rates of heat loss and 2) greater improvement of heat production. These findings suggest a functional relationship between reproductive function and seasonal thermoregulatory adjustments and indicate a significant cost to breeding during the winter months.

Adaptation, Physiological

Lasting effect of infantile cold experience on cold tolerance in adult rats.

The effect of short and repetitive exposure to cold (5 degrees C, 4 hr/day for 2 weeks) from the birth up to the 14th day of newborn rats onthe thermal regulation in adulthood and on the tolerance to cold was investigated. After being exposed to cold, they were transferred to a room at 25 degrees C (N-CA). The control rats were raised at 25 degrees C (N-WA). An acute cold exposure test was performed by placing the animals in a room at 5 degrees C under urethane anesthesia. Electrical activity of neck muscles as an index of shivering was recorded. The colonic temperature fell at a significantly slower rate in N-CA rats with less shivering than in N-WA ones. Nonshivering thermogenesis tested by norepinephrine was significantly greater in N-CA rats than in N-WA ones. These results suggest that N-CA rats developed improved cold tolerance accompanied by greater nonshivering thermogenesis. Such a phenomenon in N-CA lasted for 18 weeks after the termination of cold exposure. Adult rats subjected to the same scheme of cold exposure (A-CA) (5 degrees C, 4 hr/day, 2 weeks) showed essentially the same results as seen in N-CA, but its improved cold tolerance and elevated nonshivering thermogenesis disappeared 4 weeks after the termination of cold exposure. Extirpation of interscapular brown adipose tissue immediately before the cold test did not appreciably affect the cold tolerance in N-CA and A-CA rats. The colonic temperature at the onset of shivering was significantly lower in N-CA as well as A-CA rats than in each of the corresponding control rats, indicating a shift of the shivering threshold to lower temperature values in the animals exposed intermittently to cold. These results indicate that an infantile experience with cold results in a greater and longer sustained ability to tolerate cold in adulthood, characterized by enhanced nonshivering thermogenesis.

Acclimatization

Cold-acclimation improves cold-tolerance of diabetic rats.

1. The aim of these experiments was to study the extent to which previous cold-acclimation improves the cold-tolerance of diabetic rats. 2. Alloxan diabetic rats (fasting blood glucose higher than 200 mg/dl) were used in the experiments. 3. In Expt. 1, non-cold-acclimated control and diabetic rats were exposed to cold environment (7-9 degrees C), and the percentage of survival calculated during a 12-day experimental period. In Expt. 2, the rats were previously cold-acclimated before alloxan or saline injection (diabetic and control cold-acclimated rats) and the survival rate was also assessed during a 12-day period in the cold. 4. The percentage of survival of the non-cold-acclimated diabetic rats (Expt.1) was 19% compared with 79% of the diabetic cold-acclimated animals (Expt. 2). There were no deaths in the control groups. 5. Cold-acclimated diabetic rats maintained a near-normal thermogenic response after noradrenaline injection. This response was impaired in non-cold-acclimated diabetic rats. 6. The results of these experiments suggest that the enhanced cold-tolerance of diabetic cold-acclimated rats could be related to the increased sympathetic activity and enhanced insulin sensitivity in thermogenic tissues, such as brown fat.

Acclimatization

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