Temperature dependent redistribution of organic nitrogen during 'dry' storage of tulip bulbs cv. Apeldoorn

Tulip bulbs cv. Apeldoorn are dry stored at 5°C for 12 weeks to ensure subsequent optimal flowering when planted in the greenhouse at higher temperatures of 17–20°C. Both temperature and duration of the cold treatment determine the subsequent rate of the shoot elongation, the time until anthesis and the flower size, pigmentation and water content. In search for cold-specific physiological changes, possibly related to the development of the potential of proper flowering (flowering preparation), we studied the redistribution of organic nitrogen in both cooled (5°C) and non-cooled (17°C) bulbs.
During 12 weeks of dry storage, the total protein- and free amino acid-nitrogen content decreased in the scales, whereas the opposite was found in the basal plate (with root primordia) and the shoot. In the shoot, this occurred significantly more at 17°C than at 5°C. At the same time, there was a tissue-specific change in the free amino acid composition in both cooled and non-cooled bulbs. Changes specific for the 5°C treatment were only found for the alanine content, in both the basal plate (with root primordia) and the shoot, and for the proline, asparagine, threonine, glycine and isoleucine content, in the shoot only. These changes are, for the greater part, completed within the first 6–8 weeks of dry storage. Bulbs stored for such a short period of time at 5°C still show flowering disorders. Thus, flowering preparation is only partly accompanied by changes in free amino acid contents.     

Growth and Metabolic Activities of Heat Treated Staphylococcus aureus

Staphylococcus aureus cells which had been heated at 50 or 60° were transferred to various growth media and intracellular ribonucleic acid (RNA), deoxyribonucleic acid (DNA) and amino acids were measured during the lag phase of growth. The duration of the lag phase depended on the temperature to which the cultures had been subjected, and was longest following storage at 60°. RNA synthesis occurred almost immediately on placing treated cells in a growth medium, but at a slower rate than with unheated cells. Variation in the composition of the metabolic pool of heated cells occurred during the early lag phase and may be as a result of damage to the cytoplasmic membrane with resulting loss of permeability control.     

Priming relieves dormancy in lettuce seeds independently of changes in osmotic constituents

The relationship was studied between germination and dormancy of lettuce seeds ( Lactuca sativa L. cv. Musette) and both soluble amino nitrogen metabolism and osmotic potential. Germination at 15°C in darkness coincided with a rise in the levels of free amino acids and total soluble amino nitrogen compounds and in the activity of glutamine synthetase (GS, EC nr. 6.3.1.2). In further experiments GS activity was used as indicator of soluble amino nitrogen metabolism. GS activity increased after the start of growth indicated by an increasing intolerance to desiccation. At 30°C seeds did not germinate, unless dormancy was broken beforehand during incubation at 2° or 15°C (priming). The alleviation of dormancy occurred much earlier than the rise in the activity of GS. Priming at 15°C in polyethylene glycol instead of water retarded the breaking of dormancy and at –1.28 MPa even stimulated the induction of secondary dormancy, but did not prevent a continued rise in the activity of GS. GS activity was also not reduced during induction of secondary dormancy by dehydration of primed seeds, which antagonized the beneficial effect of priming. Psychrometric measurements showed that osmotic potential (Ψ_π) of the seeds remained constant during prolonged priming in polyethylene glycol at 15°C. During incubation in water, Ψ_π increased both prior to and after the moment of germination to less negative values. It is concluded that changes in the level of dormancy in lettuce seeds occur independently of soluble amino nitrogen metabolism and of changes in Ψ_π.     

Effect of temperature on the vegetative growth of type and field strains of Bacillus cereus

Growth of eight selected potentially pathogenic strains of Bacillus cereus was evaluated in a rich medium at different temperatures. No strain grew at 50°C; maximal growth-permissive temperatures were in the range 46–50°C for six strains and under 46°C for one strain. Faster growth occurred at 42°C. Growth may be delayed at 20°C, where the lag phase can reach 7 h. Furthermore at 20°C, cells generally show an aggregation immediately in the early exponential stage, except for two strains. Owing to this aggregation, growth is more difficult to estimate by turbidimetry at lower temperatures. These data describe the behaviour of type and field strains between 50° and 20°C and can help the prediction of shelf-life of potentially contaminated products.     

Frost tolerance in cell cultures of potato

Different methods of freezing and of estimating frost damage in cell cultures of Solanum tuberosum L. and a number of wild Solanum species were compared. Frost-killing temperatures (FKT, i.e. the temperature resulting in 50% of the maximum possible frost damage) in leaves of these species were -6°C ( S. acaule ), 5°C ( S. me-gistacrotobum ), -4.5°C ( S. commersonii ) and -3°C ( S. polytrichon and S. tubero-sum ) No appreciable species differences were found in FKT when cells were submerged in either buffer or medium and frozen. However, differences did exist when cells were frozen in a non-submerged condition: S. acaule and S. commersonii callus were more sensitive to frost than suspensions, whereas suspensions of the other species were the most sensitive. Measurement of freezing damage by either electrolyte leakage or by 2,3,5-triphenyltetrazolium chloride viability assays revealed similar FKT values. Cell cultures of S. acaule showed better frost tolerance than S. tuberosum (FKT values were -4.5 to -6°C and -2.5°C. respectively), however, frost tolerance of S. megistacrolobum and S. commersonii was only poorly expressed at the cell level (FKT values were between -2 and -3°C). Variant cell lines previously selected for resistance to the amino acid analogues hydroxyprotine, aminoethylcysteine and 5-methyhryptophan appeared to be more tolerant to frost than the wild type S. tuberosum clone.     

The effect of a self-regulating trace heating element on legionella within a shower

A self-regulating trace heating element was assessed for its ability to maintain a temperature of 50°C in the mixer valve and dead-legs of a shower, and for its effect on legionellas colonizing the shower. The trace heating element maintained a temperature of 50°C ± 1·5°C in dead-legs when the circulating hot water supply remained above 45°C. Legionellas appeared in a trace heated dead-leg when the temperature of the dead-leg reached 45°C and the hot water supply dropped below this temperature. Legionellas were eradicated or significantly reduced in sections of the shower where a temperature of 50°C was consistently achieved. The mixer valve which was trace heated but not insulated remained colonized with Legionellas. Legionellas were found in shower water throughout the study.     

Peptidase D of Escherichia coli K-12, a metallopeptidase of low substrate specificity

Abstract Peptidase D of Escherichia coli was overproduced from a multicopy plasmid and purified to electrophoretic homogeneity. The pure enzyme was stable at 4°C or −20°C and had a pH optimum at pH 9, and a p I of 4.7; the temperature optimum was at 37°C. As the enzyme was activated by Co²⁺ and Zn²⁺, and deactivated by metal chelators, it appears to be a metallopeptidase. By activity staining of native gels, 11 dipeptides which are preferentially cleaved by peptidase D were identified. Peptidase D activity required dipeptide substrates with an unblocked amino terminus and the amino group in the α or β position. Non-protein amino acids and proline were not accepted in the C-terminal position, whereas some dipeptide amides and formyl amino acids were hydrolyzed. K _m values of 2 to 5 mM indicate a relatively poor interaction of the enzyme with its substrates.     

Differential cold hardiness in adults and nymphs of the peach-potato aphid Myzus persicae

The LT₅₀ (lethal temperature) of first instar and adult stages of the peach-potato aphid Myzus persicae was lowered following long term acclimation at low temperatures.
First instars consistently showed greater cold hardiness than adult stages at each acclimation temperature, with the differential increasing as the temperature was lowered. When maintained at 5°C (the lowest acclimation regime) nymphs and adults had dLT₅₀8.3°C and 4.7°C respectively lower than those for non-acclimated individuals.
When 10°C acclimated adults were returned to 20°C, the acclimation effect was retained in full for 6 days but complete deacclimation occurred by day 10. In contrast the LT₅₀ of their progeny increased gradually from the first day of adult deacclimation towards the level of the unacclimated control over a period of 10 days.
A change in cold hardiness was observed in first instars according to their position in the birth sequence. The LT₅₀ of first-born nymphs (day 1 of reproduction) from 20°C parents was - 15.9°C rising to - 8.3°C by day 4 and remaining at this level until the end of the reproductive period.
The differential mortality between nymphs and adults observed in the laboratory was supported by the results of a field experiment. Adult aphids kept in clip-cages on a crop of oilseed rape showed greater mortality compared with those introduced as nymphs when the minimum temperature fell below -4°C for the first time in winter. At - 10°C mortality of aphids introduced as adults approached 100% whereas more than 50% of those introduced as nymphs were still alive at this temperature.     

Emergence of Baltic salmon, Salmo salar L., in relation to temperature: a laboratory study

Emergence pattern and developmental status at emergence of Baltic salmon fry from the Umeälven hatchery stock (63°50'N, 20°25'E) were studied at 6, 10 and 12°C in the laboratory. The number of days and degree days from hatching to 50% emergence decreased exponentially with increasing temperature. Synchronization of emergence increased with temperature. Optimal temperature for incubation of yolk-sac alevins was 10°C, which resulted in the largest fry at emergence and the lowest death rate. Fry kept at 6°C had the lowest mean weight and at 12°C the highest death rate. The fry emerged at an earlier developmental state with more yolk at 12°C than at 6°C. The Baltic salmon had a faster developmental rate during the gravel-phase, as compared to more southern Atlantic salmon populations.     

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 .     

一种α-环糊精葡萄糖基转移酶的纯化及性质研究

本文报道了一种主要转化产物是α-环糊精的环糊精葡萄糖基转移酶的纯化、酶学性质和转化特性。将发酵上清液通过硫酸铵分级沉淀、疏水柱层析和离子交换层析获得表观电泳纯的酶蛋白。纯酶的分子量约为75KDa,等电点5.3。最适反应温度为50℃,最适反应pH为6。对可溶性淀粉的Km值和Vmax分别是50mg/ml和6.07 mg/ml/min。色氨酸残基为酶活力的必需基团。酶的N末端序列为-SPDTSVDNKV-。Ca2+、Zn2+、Fe3+、Cu2+、Fe2+、Ag+对酶活力有强烈抑制作用。纯酶催化转化条件试验表明,廉价的马铃薯淀粉是酶催化制备α-CD的适宜底物,最佳转化条件为：酶量200u/g淀粉,温度40℃,反应时间24h,总转化率达41%,其中α-环糊精占总转化产物的78%。因此,该酶不仅表现出特殊的酶学特性,而且有较好的产业化开发前景。     

Effect of nitrogen supply on frost resistance, nitrogen metabolism and carbohydrate content in white clover (Trifolium repens)

Effects of mineral nitrogen (2, 4, 6 and 8 m M NH₄NO₃) and nodulation with Rhizobium on frost hardiness in seedlings of white clover ( Trifolium repens ) have been studied. Seedlings of a population from Bodø (67°N lat.) were grown in Leonard jars under controlled conditions in a phytotron. For induction of frost hardening, plants were first exposed to 12 h photoperiod conditions for 2 weeks at 18°C, then for 2 weeks at 6°C and finally for 2 weeks at 0.5°C. Frost hardiness after treatments at 6 and 0.5°C was significantly enhanced by increasing nitrogen supply and was positively correlated with total nitrogen content of the stolons. Frost hardiness of nodulated plants correlated to the tissue nitrogen concentration. Content of soluble proteins in stolons decreased during hardening at 6°C but did not change during treatment at 0.5°C. There were minor changes in total amount of free amino acids during hardening. Both absolute and relative amounts of proline and arginine increased, and those of asparagine decreased during hardening. Absolute amounts of all free amino acids increased with increasing nitrogen supply, but the changes during hardening were similar in all treatments. There was a significant increase in the content of soluble carbohydrates during hardening. However, this increase was inversely related to nitrogen supply.     

The type III secretion system is involved in the invasion and intracellular survival of Escherichia coli K1 in human brain microvascular endothelial cells

Two thermostable phytases were identified from Thai isolates of Aspergillus japonicus BCC18313 (TR86) and Aspergillus niger BCC18081 (TR170). Both genes of 1404 bp length, coding for putative phytases of 468 amino acid residues, were cloned and transferred into Pichia pastoris . The recombinant phytases, r-PhyA86 and r-PhyA170, were expressed as active extracellular, glycosylated proteins with activities of 140 and 100 U mL⁻¹, respectively. Both recombinant phytases exhibited high affinity for phytate but not for p -nitrophenyl phosphate. Optimal phytase activity was observed at 50 °C and pH 5.5. High thermostability, which is partly dependent on glycosylation, was demonstrated for both enzymes, as >50% activity was retained after heating at 100 °C for 10 min. The recombinant phytases also exhibited broad pH stability from 2.0 to 8.0 and are resistant to pepsin. In vitro digestibility tests suggested that r-PhyA86 and r-PhyA170 are at least as efficient as commercial phytase for hydrolyzing phytate in corn-based animal feed and are therefore suitable sources of phytase supplement.     

New distribution limits of seagrass beds in West Africa

The southern limit of the distribution of seagrass species along the west coast of Africa is not yet clearly defined. In March 2008 an expedition was organized in Senegal to search for seagrass beds from Dakar (14°45'15" N, 17°30'31" W) southwards to Tabakouta–Delta du Saloum (13°46'59" N, 16°28'42" W). Cymodocea nodosa and Halodule wrightii were observed in sandy areas of some protected bays of Dakar, around Sarène (14°16'18" N, 16°54'17" W), Joal Fadiouth (14°09'08" N, 16°50'03" W) and the Bamboung–Sourou area (13°50'08" N, 16°33'09" W), and they probably occur all along the coast of Senegal and beyond, perhaps much further south as well.     

Vasopressin and water permeability of artificial lipid membranes

Vasopressin markedly stimulated the water permeability of bilayer lipid membranes: a two-fold increase was measured at 25° in presence of 1.7·10⁻⁹ M (50 μunits/ml) vasopressin. Oxytocin and a mixture of the amino acids comprising the vasopressin molecule could not substitute for vasopressin at comparable concentration. The experimental activation energy of water transport was reduced in the presence of vasopressin from 14 to 4 kcal/mole, in agreement with the effect of the hormone on water permeability of toad bladder.     

Temperature-related fluctuations in the timing of emergence and pupation of Windermere alder-flies over 30 years

1. From 1966 to 1995, dates were recorded when adult alder-flies, Sialis lutaria L., were first seen (30-year range: 23 April – 25 May), 50% of the maximum density occurred (4 May – 4 June), and maximum density occurred (11 May – 17 June) along 200 m of Windermere shore. These emergence dates occurred at similar temperatures, estimated by mean values for both the emergence date and the week prior to emergence. The latter was the least variable at 10.1 °C (95% CL ± 0.37) for start of emergence, 11.2 °C (± 0.49) for 50% maximum density, 14.2 °C (± 0.51) for maximum density.
2. Final-instar larvae pupated in damp soil just above the water line. As laboratory temperatures were increased slowly from an initial 5 °C, the cumulative number of larvae leaving the water to pupate increased. A quadratic equation described this relationship from a threshold temperature of 7.2 °C to completion at 14.0 °C (50% point, 9.3 °C). The relationship between successful pupations and constant temperatures in the laboratory was well described by a quadratic equation with an optimum 14.9 °C (over 90% success) and no success outside the range 7–23 °C. A negative power-function described the relationship between days required for pupation and temperature, ranging from c . 28 days at 8.2 °C to c . 4 days at 22.1 °C.
3. Dates for larvae leaving the lake to pupate were back-calculated from dates for adult emergence, using the power-function for pupation time. Mean temperatures for estimated dates on which larvae left the lake to pupate were less variable than those for adult emergence, being 7.5 °C (± 0.20) for the start of pupation, 9.4 °C (± 0.16) for 50% maximum density, 13.7 °C (± 0.16) for maximum density. These values are similar to those obtained in the laboratory and can be used to predict pupation and adult emergence for different temperature regimes.     

Effect of heat treatment on the proteolytic activity of mesophilic bacteria isolated from goats' milk cheese

Strains of mesophilic lactococci and lactobacilli isolated from goats' milk cheese were grown to maximum density in milk at 30°C, pH 6·5. They were subsequently cooled to 12°C and then heated at 50°, 52° and 54°C (holding time, 15 s). The micro-organisms tested were Lactococcus lactis subsp. lactis IFPL 60, IFPL 22 and IFPL 359, Lactobacillus casei subsp. casei IFPL 731 and Lactobacillus plantarum IFPL 3, isolated from raw goats' milk cheese. The heated cells presented lower viability and acidification capacity than unheated cells. After heat treatment at 50°C, all the test strains effected practically no reduction in pH of milk (6 h), except for Lactococcus lactis subsp. lactis IFPL 60, which reduced pH to 5·9 as compared to 4·9 attained by the unheated controls. After treatment, proteolytic, aminopeptidase and dipeptidase activities of cell-free extracts decreased to a lesser extent than the number of viable cells with acidifying ability. The results suggest that these strains, if treated at 50°C, may be suitable as extra sources of important ripening enzymes in cheese making.     

Survival of the first stage larva of the metastrongyloid nematode Elaphostrongylus rangiferi under various conditions of temperature and humidity

First-stage larvae of E. rangiferi kept in water at 50°C died within 80 minutes, while at 6° the last larvae died between day 180 and 210. The time it took to reach 1_x= 0.5 (half of the larvae dead) at various temperatures between 6° and 50° was well described by the exponential function y = 614.6e^−0.15x, giving a value of 615 days to reach 1_x= 0.5 at 0°C. There was no clear decrease in the survival of larvae frozen at −20° in faeces and in water, and at −80° in faeces after 360 days. When subjected to repealed freezing and thawing, all larvae died within 77 days. When kept in air at RH = 20% and 22°C, all larvae died within 11 days, while when frozen (−20°C) in air at RH approx. 0%, 1_x stayed at approx. 0.5 from day 5 to day 16.     

INFLUENCE OF TEMPERATURE ON THE MORTALITY OF MYZUS PERSICAE (SULZER) DUE TO THE FUNGAL PATHOGEN ZOOPHTHORA PHALLOIDES BATKO

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Nymphs of Myzus persicae inoculated with Zoophthora phalloides were killed at each temperature tested in the range 10–22°C. Mortality was around 50% at 12, 15 and 18°C, but was considerably reduced outside this range. Deaths occurred soonest at 18°C (LT₅₀—6.8 days). The low temperature requirement of this fungus limits its potential as a biocontrol agent for aphids in Australia.     

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.