Changes in protein synthesis during stratification and dormancy release in embryos of sugar maple (Acer saccharum)

Protein synthesis in dormant embryos of sugar maple ( Acer saccharum ) was investigated in seeds stratified at 4°C or incubated at 15°C. Seeds stratified at 4°C germinated after 27 days; seeds incubated at 15°C failed to germinate. Stratification increased the embryo's capacity for protein synthesis by day 11 as measured by in vivo incorporation of [³⁵S]-methionine into purified protein. At 4°C protein synthesis in the embryonic axis rose in a linear fashion prior to germination, whereas in cotyledons it increased until day 20 and then declined. Analysis of radiolabelled proteins by two-dimensional gel electrophoresis revealed that the levels of specific proteins were altered by temperature, primarily in the cotyledons. Several proteins were expressed in the cotyledons at 15°C but were absent in unstratified embryos and in embryos stratified at 4°C. That is, the expression of these proteins was repressed during stratification and release from dormancy. Levels of other proteins in the cotyledons declined at 4°C during stratification. We suggest that one or more of these proteins may be associated with the inhibition of growth of the embryonic axis imposed by the cotyledons.     

Effect of temperature in the regulation of DNA synthesis in synchronous cultures of Chlorella

The influence of temperature on DNA replication of Chlorella pyrenoidosa grown at 30 °C was studied in synchronous cultures. Final DNA content and cell counts/ml were 30 to 40% less than controls after a 4 h exposure to 15 °C, but reached normal levels after being kept temporarily at 10 °C. The rates of DNA synthesis at these two temperatures were, respectively, one-sixth and one-tenth of the rate at 30 °C, but during the 15 °C treatment the cells lost the ability to enter a further replication cycle more rapidly than at 10 °C. This loss of capacity to initiate further replication cycles was prevented by inhibiting protein synthesis with cycloheximide.     

Rooting of etiolated stem segments of Populus robusta – interaction of temperature, catechol and sucrose in the presence of IAA

Etiolated stem segments of Populus robusta Schneid. were cultured in test solutions each containing 2.0 mg/1 of IAA in combination with varying concentrations of catechol or sucrose or both at 10 ± 2°C, 25 ± 1°C and 35 ± 1°C in the dark. Cuttings did not root in catechol in the presence of IAA alone at 10 ± 2°C. There was no mortality in this culture solution at this temperature. However, cuttings rooted in this culture solution at 25 ± 1°C and 35 ± 1°C and also showed mortality, which was more severe at 35 ± 1°C than at 25 ± 1°C and more severe at higher than at lower concentrations of catechol. Rooting occurred on cuttings in catechol in the presence of IAA and sucrose even at 10 ± 2°C and was strongly promoted at 25 ± 1°C and 35 ± 1°C. The mortality of segments caused by catechol was markedly lowered in the presence of sucrose at these temperatures. Sucrose thus antagonised the toxic effect of catechol.     

Effect of Iysozyme concentration, heating at 90°C, and then incubation at chilled temperatures on growth from spores of non-proteolytic Clostridium botulinum

The heat treatment necessary to inactivate spores of non-proteolytic Clostridium botulinum in refrigerated, processed foods may be influenced by the occurrence of lysozyme in these foods. Spores of six strains of non-proteolytic Cl. botulinum were inoculated into tubes of an anaerobic meat medium, to give 10⁶ spores per tube. Hen egg white lysozyme (0–50 μg ml^-1) was added, and the tubes were given a heat treatment equivalent to 19·8 min at 90°C, cooled, and incubated at 8°, 12°, 16° and 25°C for up to 93 d. In the absence of added lysozyme, neither growth nor toxin formation were observed. A 6–D inactivation was therefore achieved. In tubes to which lysozyme (5–50 μg ml^-1) had been added prior to heating, growth and toxin formation were observed. With lysozyme added at 50 μg ml^-1, growth was first observed after 68 d at 8°C, 31 d at 12°C, 24 d at 16°C, and 9 d at 25°C. Thus, in these circumstances, a heat treatment equivalent to 19·8 min at 90°C was not sufficient, on its own, to give a 6–D inactivation. A combination of the heat treatment, maintenance at less than 12°C, and a shelf-life not more than 4 weeks reduced the risk of growth of non-proteolytic Cl. botulinum by a factor of 10⁶.     

A temperature-controlled amplicon system derived from Plum pox potyvirus

The control of replication can facilitate a viral amplicon to reach high expression levels by enabling the virus to escape host defence mechanisms and reducing the deleterious effects of viral infection. We have developed a novel system to regulate amplicon expression by controlling the temperature of plant growth. Nicotiana benthamiana plants were transformed at two different temperatures with a cDNA copy of the Plum pox potyvirus genome harbouring the open reading frame 2 of Porcine circovirus 2 between the nuclear inclusion protein b and coat protein coding sequences. Although transformation at 27 °C mainly yielded nonexpressing amplicons, lines with a tight control of amplicon expression were obtained. Viral replication was not detected in these plants when germinated at 28 °C, but was observed when the plants were shifted to 20 °C. In lines transformed at 24 °C, although the amplicon was expressed at 28 °C, viral accumulation was low and caused only minor growing defects. Viral replication was enhanced in these plants by shifting the temperature to 20 °C; under such conditions, the amplicon reached higher and more persistent expression levels than in plants transformed at 27 °C. These results demonstrate the utility of temperature regulation to control viral amplicon expression.     

Effect of temperature on the hatching time of eggs of Ephemerella ignita (Poda) (Ephemeroptera:Ephemerellidae)

SUMMARY. Eggs of Ephemerella ignita (Poda) were kept at eight constant temperatures (range 5.9–19.8°C) in the laboratory. Over 85% of the eggs hatched in the temperature range 10.0–14.2°C but the percentage decreased markedly to 39% at 5.9°C and 42% at 19.8°C. Hatching time (days after oviposition) decreased with increasing water temperature over the range 5.9–14.2°C and the relationship between the two variables was well described by a hyperbola. Therefore, the time taken for development was expressed in units of degree-days above a threshold temperature. Mean values (with 95%CL) were 552 (534–573) degree-days above 4.25°C for 10% of the eggs hatched, 862 (725–1064) degree-days above 3.57°C for 50% hatched and 1383 (1294–1486) degree-days above 3.14°C for 90% hatched. These values can be used to predict hatching times at temperatures below 14.68°C for 10% hatched, 14.54°C for 50% hatched and 14.45°C for 90% hatched. At higher temperatures, the hatching time and the number of degree-days required for development both increased with increasing temperature. Equations were developed to estimate the number of degree-days required for development at these higher temperatures.
Eggs were also placed in the Wilfin Beck, a small stony stream in the English Lake District. Maximum and minimum water temperatures were recorded in each week and the summation of degree-days was used to predict the dates on which 10%, 50% and 90% of the eggs should have hatched. There was good agreement between these estimates and the actual hatching times. Only 10–15% of the eggs hatched between October and late February with most of the eggs hatching in March, April and May. Nymphs hatching in October and November probably did not survive the winter.     

Effect of Temperature on the Formation of Microsclerotia of Verticillium dahliae

Microsclerotium formation by six isolates of Verticillium dahliae was studied at different temperatures both in vitro and in Arabidopsis thaliana . In vitro mycelial growth was optimal at 25°C, but microsclerotium formation was greatest at 20°C (two isolates) or 15–20°C (one isolate). Seedlings of A. thaliana were root-dipped in a conidial suspension, planted, and either placed at 5, 10, 15, or 25°C, or left at 20°C until the onset of senescence, after which some of the plants were placed at 5, 10, 15, or 25°C. The amount of microsclerotia per unit of shoot weight was assessed in relation to isolate and temperature. The optimal temperature for production of microsclerotia was 15–25°C. Two isolates each produced about 10 times more microsclerotia than each of the other four isolates. For these isolates, high R ²_adj.-values of 0.77 and 0.66 were obtained, with temperature and its square as highly significant (P   < 0.001) independent variables. R ²_adj.-values for the other isolates varied between 0.28 and 0.39. Moving plants to different temperatures at the onset of senescence led to microsclerotial densities that were intermediate between densities on plants that had grown at constantly 20°C and plants grown at other temperatures. This suggests that vascular colonization rate and rate of microsclerotium formation are similarly affected by temperature. The senescence rate of plants appeared unimportant except for plants grown at 25°C, which showed the highest amounts of microsclerotia per unit of plant weight in the most rapidly senescing plants.     

The effect of ambient temperature on body temperature and on ultrasonic behaviour in litters of albino laboratory mice deprived of their mothers

Litters of mouse pups of various ages when deprived of their mother and kept for one hour at ambient temperatures of 3°C, 12°C and 22°C lose heat less rapidly than pups isolated individually at similar temperatures. The litters are able to maintain constant body temperatures of 35–37°C by day 8 at 22°C, by day 19 at 12°C and by day 21 at 3°C. Very little ultrasound is detected from litters of any age up to 21 days at 22°C while at 12°C ultrasound is mainly detected from pups of 4 and 6 days; very few calls are produced by older pups at this temperature. Considerably more calls are emitted at 3°C than at either of the two higher temperatures. At 3°C the total number of calls emitted during the hour-long exposure period varied markedly with age. There were two peaks of calling, one on day 6 and the other on day 16. The temporal pattern of calling throughout the hour-long exposure period at 3°C was similar in pups of 4 and 6 days and in pups of 8, 10, 12, 14 and 16 days, but differed between these two groups. In the younger group of pups the rate of calling rose to a peak and then decreased to zero as the pups became comatose; in the older pups the rate of calling was low initially then either rose throughout the rest of the hour or rose to a high level that was maintained. It is suggested that the various patterns of ultrasonic calling can be associated with the physiological development of mouse litters.     

Properties of Pseudomonads Causing Spoilage of Vegetables Stored at Low Temperature

Twenty-four strains of pectolytic, fluorescent pseudomonads were isolated from soft rots of celery stored at 0.4-1°C and five strains were isolated from soft rots in cabbage stored at 1°C. When inoculated into the vegetable from which they were isolated these bacteria caused soft rot of wounded, but not of unwounded tissue. According to their biochemical reactions, the organisms were divided into three groups; Group 1 (15 strains) were identified with Pseudomonas fluorescens Biotype II (Doudoroff & Palleroni 1974) ( Ps. marginalis ); Group 2 (12 strains) and Group 3 (two strains) would be included in the 'Miscellaneous strains'of Ps. fluorescens described by the above authors. One strain biochemically representative of Group 1 showed a maximum growth rate at 27°C (doubling time, 0.88 h) and a doubling time at 0.2°C of 14.9 h. A strain representative of Group 2 showed a maximum growth rate at 29°C (doubling time 0.96 h) and a doubling time at 0.2°C of 16.6 h. Neither strain grew at 36°C. The temperature characteristics (calculated for the range 0.2-20.8°C) were 83 011 and 79 534 J/mol, respectively. The mean doubling time for the remaining Group 1 strains at 0.2°C was 17.6 h and for remaining Group 2 strains was 17.1 h.     

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.     

Temperature control of germination and its possible role in the survival of a non-dormant population of Avena fatua

Germination of freshly harvested seeds of a non-dormant (ND) line (Stonehouse 319) of wild oats ( Avena fatua L.) was inhibited by incubation of the seeds at relatively high temperatures of 25 and 30°C. The germination inhibition in these seeds appeared to be a case of thermo-inhibition which was the direct effect of hightemperature treatment (HIT), since it did not persist after transferring the seeds to an optimum germination temperature of 20°C. Even a prolonged HTT of 30°C for over 5 weeks did not prevent germination of about 80% of the seeds transferred to 20°C. However, in a significant proportion of the seeds, thermo-dormancy was induced by 10 days of HTT at 30°C if the seeds were then incubated at sub-optimal temperatures of 5 to 15°C. This thermo-dormancy would appear to be 'restrictive' in form, since its expression was restricted to very specific conditions. Relatively low inclubation temperaturs of 5 and 10°C markedly slowed germination whether HTT was applied or not. The results suggest that thermo-inhibition and thermo-dormancy, induced during seasonal temperature fluctuations, may provide a survival mechanism for seeds of such ND lines as Stonehouse 319.     

The effect of temperature on growth and reproduction of Scytosiphon complanatus (Fucophyceae) from Greenland

Scytosiphon complanatus from Greenland was grown under long-day conditions on a temperature-gradient device with a temperature range from 5.4°C to 31.8°C. Growth was optimal between 16.0°C and 20.9°C. In a four week experimental period at 5.4°C and 7.5°C growth was slow and not measurable. The inoculated germlings died at temperatures between 24.0°C and 27.5°C. Under all temperatures the prostrate systems, knot filaments or ralfsioid thalli, as well as the parenchymatous macrothalli remained sterile during the experimental period. Prolongation of the growth period showed that formation of swarmers was prevented at temperatures above 18.6°C. The geographic distribution is discussed in relation to these results.     

Effect of pre- and post-heat shock temperature on the persistence of thermotolerance and heat shock-induced proteins in Listeria monocytogenes

The effect of incubation temperature, before and after a heat shock, on thermotolerance of Listeria monocytogenes at 58°C was investigated. Exposing cells grown at 10°C and 30°C to a heat shock resulted in similar rises in thermotolerance while the increase was significantly higher when cells were grown at 4°C prior to the heat shock. Cells held at 4°C and 10°C after heat shock maintained heat shock-induced thermotolerance for longer than cells held at 30°C. The growth temperature prior to inactivation had negligible effect on the persistence of heat shock-induced thermotolerance. Concurrent with measurements of thermotolerance were measurements of the levels of heat shock-induced proteins. Major proteins showing increased synthesis upon the heat shock had approximate molecular weights of 84, 74, 63, 25 and 19 kDa. There was little correlation between the loss of thermotolerance after the heat shock and the levels of these proteins. Thermotolerance of heat shocked and non-heat shocked cells was described by traditional log-linear kinetics and a model describing a sigmoidal death curve (logistic model). Employing log-linear kinetics resulted in a poor fit to a major part of the data whereas a good fit was achieved by the use of a logistic model.     

The cell kinetics of teleost fish epidermis: Epidermal mitotic activity in relation to wound healing at varying temperatures in plaice (Pleuronectes platessa)

An autoradiographic study of experimental wounding of the skin of juvenile plaice was performed using tritiated thymidine (20 μCi per gram body weight) injected intraperitoneally, at water temperatures of 5, 10 and 15°C.
Samples of lesion were taken at regular intervals up to 108 hours and autoradiographic preparations made. In addition wounds of less than 12 hours duration, unsuitable for autoradiography were examined and in vitro explant autoradiography studies on similar material used to support the findings for these short term lesions. Results showed that there was a very rapid migration of Malpighian cells into the defect which was not accompanied by any evidence, over the time-scale of the study, of a mitotic burst. Adjacent normal skin was markedly reduced in thickness. Closure of these relatively small wounds was achieved within nine hours at 10°C and by 12 hours at 5°C. The thickness of the migrated epidermal cover was much thicker at 15°C than 5°C.
Numbers of labelled cells were similar in migrating and peripheral epidermis and mucous cells appeared to be randomly distributed except in the periphery of advancing migrating cells, where thev were absent.     

Effect of temperature on the demography of Galerucella birmanica (Coleoptera: Chrysomelidae)

Studies on the effect of temperature on the development of the water chestnut beetle, Galerucella birmanica Jacoby were carried out in the laboratory at seven different temperatures: 16 °C, 19 °C, 22 °C, 25 °C, 28 °C, 31 °C and 34 °C. The developmental time decreased with increase in temperature. The developmental time at 16 °C, 19 °C, 22 °C, 25 °C, 28 °C, 31 °C and 34 °C was 96.60, 80.68, 58.96, 43.48, 35.03, 30.08 and 28.02 days for the period from egg hatching to adult emergence, respectively. The developmental threshold estimated for a generation by linear regression was 10.36 °C. The fecundity per female at 22 °C, 25 °C, 28 °C, 31 °C and 34 °C was 102.3, 134.5, 141.2, 130.1 and 116.2 eggs, respectively. Oviposition period ranged from 15.6 days at 22 °C to 8.6 days at 34 °C. Hatchability of eggs was highest at 31 °C with 76.9% and lowest at 34 °C with 57.1%. The highest generation survival rate was 65.3% at 31 °C, and the intrinsic rate of natural increase ( r _m) for G. birmanica was the highest at 34 °C.     

Interactions of temperature and photoperiod in the control of flowering of latitudinal and altitudinal populations of wild strawberry (Fragaria vesca)

Floral induction and development requirements of a range of latitudinal and altitudinal Norwegian populations of the wild strawberry Fragaria vesca L. have been studied in controlled environments. Rooted runner plants were exposed to a range of photoperiods and temperatures for 5 weeks for floral induction and then transferred to long day (LD) at 20°C for flower development. A pronounced interaction of temperature and photoperiod was shown in the control of flowering. At 9°C, flowers were initiated in both short day (SD) and LD conditions, at 15 and 18°C in SD only, whereas no initiation took place at 21°C regardless of daylength conditions. The critical photoperiod for SD floral induction was about 16 h and 14 h at 15 and 18°C, respectively, the induction being incomplete at 18°C. The optimal condition for floral induction was SD at 15°C. A minimum of 4 weeks of exposure to such optimal conditions was required. Although the populations varied significantly in their flowering performance, no clinal relationship was present between latitude of origin and critical photoperiod. Flower development of SD-induced plants was only marginally advanced by LD conditions, while inflorescence elongation and runnering were strongly enhanced by LD at this stage. The main shift in these responses took place at photoperiods between 16 and 17 h. Unlike all other populations studied, a high-latitude population from 70°N ('Alta') had an obligatory vernalization requirement. Although flowering and fruiting in its native Subarctic environment and after overwintering in the field in south Norway, this population did not flower in the laboratory in the absence of vernalization, even with 10 or 15 weeks of exposure to SD at 9°C. Flowering performance in the field likewise indicated a vernalization requirement of this high-latitude population.     

The Effects of Cooling Conditions on the Behavior of Oxytricha bifaria (Ciliophora Hypotrichida)

The new analytic approach to the behavior of ciliates represented by the ethogram was used to study the locomotion of Oxytricha bifaria at different temperatures, isotropically applied according to a new protocol. It was shown that under these experimental conditions as the temperature dropped in stages from 24°C to 19°C to 14°C to 9°C, the general mobility of experimental populations decreased, as indicated by 1) the decreasing percentage of mobile organisms, 2) their decreasing velocity, 3) their prolonged backward creeping, and 4) the increasing length of their immobilization periods. The ethogram more particularly revealed that decreasing temperatures induced 1) the appearance of rightward arcs (several of them being travelled by specimens sliding on the substrate). 2) the reduction of both the radius and the velocity along the normal leftward arcs (A-) and the segments (S), and 3) the symmetric increase of both the central angle and the duration of the A- and S. These changes were reversibly induced: they disappeared when the temperature returned to 24°C. Moreover. a new behavioral pattern, the prolonged Side Stepping Reaction, was found. Helicoidal swimming, occurring only at 14°C, was analyzed. Among all of the behavioral parameters, nine were shown to change dramatically between 14°C and 9°C, demonstrating that the linear cooling of the populations induced clear non-linear effects.     

Bimodal life cycle of the burying beetle Nicrophorus quadripunctatus in relation to its summer reproductive diapause

Abstract 1. Under natural conditions in Kyoto, Japan, the reproductive activities of Nicrophorus quadripunctatus Kraatz (Coleoptera: Silphidae) decreased in summer and the species showed a bimodal life cycle.
2. In the laboratory, most adult pairs raised at 20 °C under a LD 12:12 h regime reproduced when provided with a piece of chicken. In adults raised at 20 °C under a LD 16:8 h regime, however, both reproductive behaviour and ovarian development were reduced. It is concluded that these adults entered a reproductive summer diapause.
3. High temperature (25 °C) also suppressed the reproductive behaviour even under a favourable LD 12:12 h regime. In the field, therefore, adults reduce their reproductive activity in summer because of diapause induced by long-day photoperiods and direct inhibition of reproduction by high temperatures.
4. When the temperature was changed from 20 °C to 25 °C immediately after hatching of larvae, they reached the wandering stage in 95% of adult pairs. When the temperature was changed from 20 °C to 25 °C immediately after oviposition, however, no larvae hatched in 85% of pairs. Egg mortality was significantly higher at 25 °C than at 20 and 22.5 °C; no eggs hatched at 27.5 °C. The physiological mechanisms for reducing reproduction probably prevent the beetles from inefficient oviposition in summer.     

Generation of a membrane-bound, oligomerized pre-pore complex is necessary for pore formation by Clostridium septicum alpha toxin

Low-temperature inhibition of the cytolytic activity of alpha toxin has facilitated the identification of an important step in the cytolytic mechanism of this toxin. When alpha toxin-dependent haemolysis was measured on erythrocytes at various temperatures it was clear that at temperatures ≤15°C the haemolysis rate was significantly inhibited with little or no haemolysis occurring at 4°C. Alpha toxin appeared to bind to and oligomerize on erythrocyte membranes with similar kinetics at 4°C and 37°C. The slight differences in these two processes at 4°C and 37°C could not account for the loss of cytolytic activity at low temperature. At 4°C alpha toxin neither stimulated potassium release from erythrocytes nor formed pores in planar membranes. In contrast, at temperatures ≥25°C both processes proceeded rapidly. Pores that were opened in osmotically stabilized erythrocytes could not be closed by low temperature. Therefore, low temperature appeared to prevent the oligomerized complex from forming a pore in the membrane. These data support the hypothesis that alpha toxin oligomerizes into a membrane-bound, pre-pore complex prior to formation of a pore in a lipid bilayer.