Effect of long-term and rapid cold hardening on the cold torpor temperature of an aphid

Abstract.  The effect of long-term (seasonal) acclimation and rapid cold hardening is investigated on the cold torpor temperature ( CT _min) of adult grain aphids, Sitobion avenae, reared at 20 or 10 °C for more than 6 months before experimentation. Rapid cold hardening is induced by exposing aphids reared at 20 to 0 °C for 3 h and aphids reared at 10 to 0 °C for 30 min (acclimation regimes previously found to induce maximum rapid cold hardening). The effect of cooling aphids from the same rearing regimes from 10 to −10 °C at 1, 0.5 and 0.1 °C min⁻¹ is also investigated. In the 20 °C acclimated population, rapid cold hardening and cooling at 0.1 °C min⁻¹ both produce a significant decrease in CT _min from 1.5 ± 0.3 to –0.9 ± 0.3 and –1.3 ± 0.3 °C, respectively. Rapid cold hardening also results in a significant reduction in CT _min of the population reared at 10 °C from 0.8 ± 0.1 to –0.9 ± 0.2 °C. However, none of the cooling regimes tested reduces the CT _min of the winter-acclimated (10 °C) population. The present study demonstrates that rapid cold-hardening induced during the cooling phase of natural diurnal temperature cycles could lower the movement threshold of S. avenae , allowing insects to move and continue feeding at lower temperatures than would otherwise be possible.     

Rapid cold-hardening in a Karoo beetle, Afrinus sp.

Abstract.  In the insect rapid cold-hardening response, survival at subzero temperatures is greatly improved by a brief pre-exposure at a milder temperature. It is predicted that insects with minimal cold tolerance capabilities living in variable environments should use rapid cold-hardening to survive sudden cold snaps. This is tested in Afrinus sp., a beetle that lives in an exposed habitat on rock outcrops in the Karoo Desert, South Africa, where microclimate temperatures drop infrequently to below freezing. Afrinus sp. shows a significant rapid cold-hardening response: survival of a 2-h exposure to −6.5 °C is much improved after pre-exposure to −2 °C, to 0 °C with a 2-h return to the rearing temperature, and to 40 °C, but not after pre-exposure to 0 °C. Little is known about the mechanism of the rapid cold-hardening response, although the data suggest that rapid cold-hardening may be mediated via several different mechanisms.     

Cold tolerance in tomato. I. Seed germination and early seedling growth of Lycopersicon esculentum

Tomato seed germination times were evaluated foi three "cold germinating" Lycopersicon esculentum Mill, accessions, PI 120256, PI 174263 and PI 341988 and a control breeding line, T3, at temperatures of 6 to 20°C. Accelerated failure analysis indicated that although PI 120256, 174263 and 341988 germinated more rapidly than T3 from 20 to 9°C, the minimum temperatures for germination were similar, and germination times of PI 120256 and 341988 were relatively more inhibited by progressively lower temperatures than was T3. Rapid germination of these three Pls at 10°C may not be due to cold tolerance, but to seed characteristics that promote rapid germination. Hypocotyl and root elongation over time were described by a three-parameter logistic equation; the growth rate parameter for hypocotyl elongation of all four genotypes was greatly inhibited from 20 to 15 and 10°C. Multivariate and univariate analyses of hypocotyl growth parameters indicated significant differences among accessions, but no significant genotype by temperature interaction. Rapid emergence reported for these Pis at 10°C is attributable to early germination, rather than rapid hypocotyl growth.     

The influence of thermal parameters on the acclimation responses of pinfish Lagodon rhomboides exposed to static and decreasing low temperatures

Pinfish Lagodon rhomboides acclimation rates were determined by modelling changes in critical thermal minimum ( T _{crit min}, ° C) estimates at set intervals following a temperature decrease of 3–4° C. The results showed that pinfish gained a total of 3·7° C of cold tolerance over a range of acclimation temperatures ( T _acc, ° C) from (23–12° C), that cold tolerance increased with exposure time to the reduced temperature at all T _acc, but that the rate of cold tolerance accruement (mean 0·14° C day⁻¹) was independent of T _acc. A highly significant ( P < 0·001) multivariate predictive model was generated that described the acclimation rates and thermal tolerance of pinfish exposed to reduction in water temperature: log₁₀ T _{crit min}= 0·41597 − 0·01704 T _acc+ 0·04320 T _plunge− 0·08376[log₁₀ ( t + 1)], where T _plunge is plunge temperature (° C) and t is the time (days). A comparison of the present data, with acclimation rate data for other species, suggests that factors such as latitude or geographic range may play a more important role than ambient temperature in determining cold acclimation rates in fishes.     

Impact of Corticotropin-Releasing Hormone on Extracellular Norepinephrine in Prefrontal Cortex After Chronic Cold Stress

Abstract: We have previously demonstrated that exposing rats to cold (5°C) for 3–4 weeks potentiates the increase in extracellular norepinephrine (NE) in the medial prefrontal cortex produced by acute tail shock. In the present study, we used microdialysis to determine the duration of cold exposure required to produce this sensitization and explored the mechanism of the phenomenon. Tail shock elicited a twofold greater increase in extracellular NE in the medial prefrontal cortex of rats exposed to cold for 2 weeks than in naive control rats or in rats exposed to cold for 1 week and tested either immediately or after a 2-week delay. Local infusion of 10 µ M d -amphetamine or 30 m M K⁺ increased extracellular NE in the medial prefrontal cortex (∼350 and 190%, respectively) comparably in control rats and rats exposed to cold for 3 weeks. In contrast, intraventricular administration of 3.0 µg of corticotropin-releasing hormone increased extracellular NE in the medial prefrontal cortex by 65% in rats exposed to cold for 2 weeks, but only 35% in control rats. These results indicate that an enhanced responsiveness of noradrenergic neurons to acute tail shock (1) requires ∼2 weeks of cold exposure to develop and (2) may be mediated by a change at the level of the noradrenergic cell bodies rather than the nerve terminals.     

The impact of fluctuating thermal regimes on the survival of a cold-exposed parasitic wasp, Aphidius colemani

Abstract.  The impact of fluctuating thermal regimes on the cold tolerance of the parasitoid Aphidius colemani at the mummy stage is examined. The hypothesis is tested that, if a cold period is interrupted by a return to a higher temperature for a short time, a physiological recovery is possible and may lead to higher survival. Mummies are exposed to a constant temperature of 4 °C and, when periodic sudden transfers to 20 °C for 2 h are applied, survival of immature parasitoids is markedly improved, proportionally to the warming frequency. The time lag before emergence diminishes with the duration of cold exposure and with warming frequency. The sex ratio of emergent adults after cold exposure indicates that males may be more susceptible than females to cold-injury. It is suggested that the transfer to 20 °C allows a re-activation of the normal metabolism, leading to repair and recovery of any injuries caused by prolonged chilling. The study underlines the importance of cyclic temperature changes on insect survival.     

Potato cold hardiness development and abscisic acid. II. De novo synthesis of proteins is required for the increase in free abscisic acid during potato (Solanum commersonii) cold acclimation

During cold acclimation of potato plantlets ( Solanum commersonii Dun, PI 458317), there are two transitory increases in free ABA content corresponding to a three-fold increase on the 2nd day and a five-fold increase on the 6th day (Ryu and Li 1993). During this period, plantlets increased in cold hardiness from −5°C (killing temperature, control grown at 22/18°C, day/night) to −10°C by the 7th day of exposure to 4/2°C (day/night). This increase in free ABA was not found when cycloheximide (CHI), an inhibitor of cytoplasmic protein synthesis, was added to the culture medium 6 h before exposure to low temperatures. Plantlets treated with CHI did not acclimate to cold, maintaining a hardiness level (−5°C) similar to that of the 22/18°C-grown plantlets. When the CHI-treated plantlets were exposed to low temperatures for 3 days, transferred to CHI-free culture medium and grown at low temperatures, the plantlets showed a transitory increase in free ABA 2 days later. This increase was followed by the development of cold hardiness (−8°C). Application of CHI to the culture medium after 3 days of cold acclimation, when the first ABA peak and a partial development of cold hardiness (−8°C) had occurred, blocked the second transitory increase in free ABA and resulted in no further development of cold hardiness. These results suggest that de novo synthesis of proteins is required for these transitory increases in free ABA during cold acclimation of potato plantlets.     

Cold shock and cold tolerance in larvae and pupae of the blow fly, Lucilia sericata

Abstract.  Understanding the effects of low winter temperatures on mortality is essential in the development of a full understanding of the long-term population dynamics of any insect. The present study aims to examine the survival of pupae and larvae of the blow fly, Lucilia sericata , at overwintering temperatures. Groups of pupae and diapausing and nondiapausing third-stage larvae of L. sericata are maintained in cooled incubators at either 3 °C and 6 °C. Groups are removed from the incubators at 3–4-day intervals and transferred either to−8 °C or to 25 °C. After 1 h in the freezer, the larvae and pupae exposed to this cold-shock are also transferred to 25 °C. Larvae and pupae are then allowed to continue development and the number of adults emerging from each group is counted. The results demonstrate that survival decreases linearly with the period of exposure at both 3 °C and 6 °C. Mortality is higher at 3 °C than at 6 °C and, in groups that receive the cold shock, cold-shock reduces emergence by over 50%. However, there is no consistent tendency for diapausing larvae to survive prolonged cold or cold shock better than other life-cycle stages. The results suggest that the facultative development of an overwintering diapause stage in L. sericata does not appear to be an adaptation to enhance cold tolerance or resistance to cold shock. It is concluded that the survival of overwintering L. sericata is likely to be relatively less affected by low temperatures than it is by, for example, biotic factors, particularly given the buffered soil environment and short time-scales over which periods of cold act.     

Intergenerational acclimation in aphid overwintering

Abstract.  1. When first instar nymphs and adults of the grain aphid Sitobion avenae (Fabricius) (Hemiptera: Aphidiae) were maintained in long-term cultures (>6 months) at 20 °C and 10 °C, the LT₅₀ decreased from −8 and −8.8 °C to −16.0 and −13.5 °C, respectively.
2. When aphids from the 20 °C culture were transferred to 10 °C, there was a progressive increase in cold tolerance through three successive generations. Transfer of newly moulted pre-reproductive adults reared at 10 °C for three generations back to 20 °C resulted in a rapid loss of cold hardiness in their nymphal offspring.
3. In all generations reared at 10 °C, first born nymphs were more cold hardy than those born later in the birth sequence. The LT₅₀ of nymphs produced on the first day of reproduction in the first, second and third generations maintained at 10 °C were −14.8, −17.0 and −16.6 °C, respectively. Thereafter, nymphal cold hardiness decreased over the subsequent 14 days of reproduction in each generation at 10 °C with mean LT₅₀ values of −10.3, −12.6 and −14.8 °C, respectively. By contrast, the cold tolerance of first born nymphs of aphids reared continuously at 20 °C did not differ in comparison with later born siblings. The LT₅₀ of adult aphids was also unaffected by ageing.
4. The ecological relevance of these findings is discussed in relation to the overwintering survival of aphids such as S. avenae .     

Effect of the exposure to chronic-intermittent cold on the thyrotropin and thyroid hormones in the rat

Rats exposed to acute cold (4 degrees C for 2 h), chronic cold (4 degrees C), and chronic-intermittent cold (4 degrees C for 2 h daily) were killed after 1, 2, 3, 4, and 10 days of cold exposure. The control group was maintained at 25 degrees C. In each animal, the plasma concentration of thyrotropine (THS), triiodothyronine (T3), and thyroxine (T4) was determined by radioimmunoassay. At the initial time of exposure, elevations in TSH, T3, and T4 were observed in the rats in each experimental group. However, on the 10th day, in rats exposed to chronic-intermittent cold, TSH, T3, and T4 decreased to values lower than the control values. In animals exposed to acute cold as well as to chronic cold no differences were found, with respect to the controls, in TSH and T4. In rats exposed to acute cold for 10 days, the T3 value was lower than the control value; however, in animals exposed to chronic cold, T3 was same as that in the controls. The results indicate that, in the rat, exposure to chronic-intermittent cold produces an inhibition in the secretion of TSH and thyroid hormones.     

Oxygen consumption, ammonia excretion and protein use in response to thermal changes in juvenile Atlantic salmon Salmo salar

Experiments were designed to examine the effects of various temperature challenges on oxygen consumption and ammonia excretion rates and protein utilization in juvenile Atlantic salmon Salmo salar . Fish acclimated to 15° C were acutely and abruptly exposed to either 20 or 25° C for a period of 3 h. To simulate a more environmentally relevant temperature challenge, a third group of fish was exposed to a gradual increase in temperature from 15 to 20° C over a period of 3 h ( c. 1·7° C h⁻¹). Oxygen consumption and ammonia excretion rates were monitored before, during and after the temperature shift. From the ammonia excretion and oxygen consumption rates, protein utilization rates were calculated. Acute temperature changes (15–20° C or 15–25° C) caused large and immediate increases in the oxygen consumption rates. When the temperature was gradually changed ( i.e. 1·7° C h⁻¹), however, the rates of oxygen consumption and ammonia excretion were only marginally altered. When fish were exposed to warmer temperatures ( i.e. 15–20° C or 15–25° C) protein use generally remained at pre-exposure (15° C) levels. A rapid transfer back to 15° C (20–15° C or 25–15° C) generally increased protein use in S. salar . These results indicate that both the magnitude and the rate of temperature change are important in describing the physiological response in juvenile salmonids.     

Temperature- and developmentally-induced variation in the histochemical profile of myofibrillar ATPase activity in carp

The histochemical profile of calcium activated acid stable myofibrillar ATPase (mATPase) activity in developing larval and juvenile carp was investigated. In the larval fish, differentiation of pink muscle fibres occurred after metamorphosis which was delayed by a week at 17° C compared to larvae grown at 27° C. After metamorphosis the 27° C group exhibited some small myofibres with acid stable mATPase activity in the deep white muscle. This was similar for the juvenile carp which were acclimated for more than a month at 25° C. In contrast, the cold (12° C) acclimated juvenile fish, contained very few small white muscle fibres with acid stable mATPase activity. It was also noted that the cold acclimated fish had lower background acid stable mATPase activity than the warm acclimated fish. Results indicate that after metamorphosis and more evidently in juveniles, temperature can influence the rate of myofibre hyperplasia.     

Geographical variation in egg cold hardiness: a study on the adaptation strategies of the migratory locust Locusta migratoria L.

Abstract. 1. For many species of insect, cold hardiness is an important trait that enables a population to develop in the next season and to extend its range. To elucidate the role of cold hardiness of the migratory locust Locusta migratoria L. in its outbreak and distribution areas, egg cold hardiness was examined in locusts derived from four locations from latitude 18°23'N to latitude 41°10'N in eastern China.
2. The supercooling points of eggs from different geographic populations did not differ significantly for the first development stage, with an average ± SE of −24.5 ± 0.51 °C, or for the second stage, −22.06 ± 0.68 °C, however there was a significant difference for the embryonic development phase among the four geographical populations. The egg supercooling point increased gradually from neonatal egg to old egg; eggs prior to hatching always had a much higher supercooling point.
3. Comparisons of the cold hardiness of four populations were carried out by validating the close correlation between latitude and the effects of cold on hatching, low lethal temperature (Ltemp₅₀), and low lethal time (Ltime₅₀). There were significant differences among the four populations; the northern population was more cold hardy than the southern population, and the two mid-latitude populations were intermediately cold hardy.
4. The cold hardiness of all populations was enhanced to various degrees by short-term cold acclimation at 0 °C and 5 °C. For most populations, a 2-day acclimation period seemed to be optimal.     

Effects of cold (+4°C) on thyroid metabolism of iodine (I and I) in quail (Coturnix coturnix japonica)—A comparison with the rat

1. 1.|A kinetic study of the absolute specific activity (SA) of iodine in intrathyroidal iodotyrosines and iodothyronines, and in plasma PBI, was performed comparatively in adult male rats and quail kept at +25°C (controls) or exposed to +4°C (for 2 days or 30 days before the iodine measurements).

2. 2.|In both species, 2 days of cold exposure induced a rapid increase (20 to 50%) in the SA of intrathyroidal iodoaminoacids and plasma PBI, leading to a rapid equilibration between the SA measured. After 30 days an acceleration in iodine turnover was still measurable in rats, but the stimulation then appeared more important in quail. In addition, the acceleration of iodine turnover led to a characteristic bimodal pattern of this turnover which was apparent only in quail, and could reflect an important production of non-hormonal iodinated proteins. Although the bird thyroid has been claimed to be less reactive to cold than its mammalian homologues, the present study indicates that, at least in bird species whose thyroidal stores of iodine are not excessive, the thyroid is even more responsive to cold than the rat thyroid.

Induction of embryos in pollen cultures of Nicotiana sylvestris

Induction of embryos in pollen cultures was possible when young buds from plants induced to flower at 15°C were given a cold treatment at 10°C for 15 days. Following selective isolation of embryogenic pollen the cultures raised were given cold treatment at 10°C for 12 or more days. In such cultures the embryos appeared at a frequency of 1.2% of the cultured pollen on mineral-sucrose medium at pH 6.8.     

Effects of temperature and moisture stress on the accumulation of abscisic acid in bean

Levels of abscisic acid (ABA) and several indicators of leaf-water status were measured in excised and intact primary leaves of bean ( Phaseolus vulgaris cv. Kinghorn) exposed to different temperature and moisture regimes. After 2 h at 5°, 25° or 45°C under moist conditions, the water status of detached leaves showed only minor changes, and there was no increase in ABA. Under conditions favoring water loss, ABA rose over 10-fold at 25°C, and trends towards higher ABA levels were observed at 5° and 45°C. When intact leaves on whole plants were exposed to the same temperatures for 10 h, there was still no evidence of a temperature-dependent rise in ABA that was not associated with a disturbance in the water balance of the plant. These data suggest that the rapid accumulation of ABA during temperature stress is a function of induced moisture deficits and does not result from high or low temperatures per se.     

Temporal resolution of cold acclimation and de-acclimation in the Antarctic collembolan, Cryptopygus antarcticus

Abstract.  The Antarctic collembolan, Cryptopygus antarcticus (Willem), can switch its supercooling point (SCP) between 'winter' and 'summer' modes of cold hardiness over a matter of hours. High resolution temporal scaling of the acquisition and loss of cold hardiness is undertaken by assaying changes in the proportion of animals freezing below −15 °C in response to cooling rate, acclimation temperature, and access to food and moisture. Rapid de-acclimation to the 'summer' modal state is readily achieved after 1–6 h in response to warming and access to food; however, rapid acclimation to the 'winter' modal state is only evident in response to slow cooling and narrow ranges of temperature (0–5 °C). The rapid loss of cold tolerance at higher temperatures with access to food, in particular, emphasizes this species' opportunistic responses to resource availability in the short polar summers. Cold hardiness is apparently more readily traded off against nutrient acquisition than vice versa in this maritime Antarctic species.     

Stress-Induced Sensitization of Dopamine and Norepinephrine Efflux in Medial Prefrontal Cortex of the Rat

Abstract: We examined whether prior exposure to chronic cold (17–28 days, 5°C) alters basal or stress-evoked (30-min tail shock) catecholamine release in medial prefrontal cortex, nucleus accumbens, and striatum, using in vivo microdialysis. Basal norepinephrine (NE) concentrations in medial prefrontal cortex did not differ between chronically cold-exposed rats and naive control rats (2.7 ± 0.3 vs. 2.5 ± 0.2 pg/20 µl, respectively). Basal dopamine (DA) efflux in any of the brain regions was not significantly different between chronically cold-exposed rats and naive rats. However, a trend for lower basal DA efflux in the cold-exposed relative to naive rats was observed in medial prefrontal cortex (1.5 ± 0.2 vs. 2.2 ± 0.3 pg/20 µl, respectively), nucleus accumbens (3.7 ± 0.8 vs. 5.4 ± 0.9 pg/20 µl, respectively), and striatum (4.4 ± 0.5 vs. 7.2 ± 1.5 pg/20 µl, respectively). In medial prefrontal cortex of rats previously exposed to cold, tail shock elicited a greater increase from baseline in both DA and NE efflux relative to that measured in naive rats (DA, 2.3 ± 0.3 vs. 1.2 ± 0.1 pg, respectively; NE, 3.8 ± 0.4 vs. 1.4 ± 0.2 pg, respectively). However, in nucleus accumbens or striatum of rats previously exposed to cold, the stress-induced increase in DA efflux was not significantly different from that of naive rats (nucleus accumbens, 1.8 ± 0.7 vs. 1.5 ± 0.3 pg, respectively; striatum, 1.9 ± 0.4 vs. 2.6 ± 0.7 pg, respectively). Thus, both cortical NE projections and cortically projecting DA neurons sensitize after chronic exposure to cold. In contrast, subcortical DA projections do not sensitize under these conditions.     

Contact with the host plant enhances aphid survival at low temperatures

1. The hypothesis, suggested by previous studies, that host plant contact reduces the cold tolerance of anholocyclic aphids was tested under laboratory conditions. Adult and first-instar Rhopalosiphum padi (L.) were exposed to temperatures of 0, –5 and –10 °C on intact plants, excised leaves and in the absence of contact with plant material.
2. Median lethal time (LT₅₀) values at all three temperatures indicate that aphids exposed in association with plant material survive longer than aphids that have no contact with their host plant. The difference in survival was most pronounced at –10 °C. Therefore, the above hypothesis is rejected for aphids on cereals because host plant contact apparently enhances cold tolerance.
3. Exposure on excised leaves also enhanced aphid survival at low temperature but was less effective than the intact plant. This suggests that plant quality as well as the presence or absence of plants is important in the cold tolerance of aphids on cereals.     

Homeostatic regulation of acetazolamide-sensitive esterase activity in the bluegill sunfish, Lepomis macrochirus, following acute cold shock

Acetazolamide-sensitive esterase activity was elevated in branchial homogenates of control juvenile bluegill sunfish, Lepomis macrochirus , acclimated at 20° C but decreased rapidly within 9 h following an acute hypothermal shock to 8° C. After 2 weeks at 8° C, shocked-fish enzyme activity was similar to control fish. At 20° C acclimation temperature, specific activity of bluegills was similar in swimbladder, liver, kidney, gill, spleen, and gonad homogenates and was significantly higher (α=0.05 level) in whole blood homogenates. The pH optima for enzymes extracted from fish acclimated at 20° and 8° C were 7.29 and 8.00, respectively. Polyacrylamide gel electrophoresis (PAGE) demonstrated two distinct forms of acetazolamide-sensitive esterase activity present in both 20° and 8° C acclimated fish. Specific activity for homogenates from both 20° and 8° C acclimated fish differed significantly when assayed at 20° C, suggesting both qualitative and quantitative changes in acetazolamide-sensitive esterase. It is postulated that relatively rapid alterations in esterase activity promote survival in bluegill following acute cold shock through the central role of enzymes in the regulation of plasma ion concentrations and acid/base equilibria.