The influence of temperature on light enhanced glycoalkaloid synthesis in potato

The influence of temperature on total glycoalkaloid (TGA) synthesis in tubers exposed to light (250 jumol m“² s”² PAR, Photosynthetically Active Radiation) or dark environments for 96 h was examined in three potato cultivars. Following 96 h light or dark the tubers were stored without light at 5°C or 24°C and TGA concentrations monitored over the subsequent 30 and 90 days. Exposure to light and cultivar were found to be major factors influencing TGA concentrations; temperature had no significant effect. TGA content in illuminated tubers of cvs ‘Pentland Hawk’ and ‘Kerrs Pink’ were significantly higher (P < 0.01) compared with tubers placed in the dark. TGA concentrations in cv. ‘Desiree’ increased significantly only following exposure to light at low temperatures (P < 0.05). Removal of tubers from storage at 5°C and immediate illumination at 24°C altered the ratio of glycoalkaloids in cvs ‘Pentland Hawk’ and ‘Kerrs Pink’. Regardless of cultivar and storage temperature TGA concentrations were higher at the end of the storage period compared with initial TGA concentrations. During storage TGA concentrations fluctuated widely and gradual accumulation of glycoalkaloids with time was rarely demonstrated except in cv. ‘Desiree’. Tubers stored at 24°C accumulated higher TGA concentrations than those stored at 5°C in cv. ‘Kerrs Pink’ but not in cvs ‘Pentland Hawk’ and ‘Desiree’. Tubers of cv. ‘Kerrs Pink’ exposed to light prior to storage accumulated glycoalkaloids more rapidly than unexposed tubers during storage at 24°C and occasionally at 5°C. Light enhanced glycoalkaloids are not degraded over time.     

Effects of temperature on hatching rate, embryonic development and early larval survival of the edible sea urchin, Tripneustes gratilla

The temperature tolerances of embryonic and early larval development stages of Tripneustes gratilla were investigated from 13-34°C under laboratory conditions. Zygotes showed unequal cleavage at 13°C, whereas cleavage did not occurred at 34°C. Hatching was observed between 16–31°C with maximum hatching rates observed at 22–29°C. The lower and higher temperature limits for embryonic development were approximately 22°C and 29°C, respectively. Outside of this temperature range, embryos showed abnormality at different incubation times. Early larvae of this species have the ability to survive the higher temperature limit for short periods of time. Prism and 2 arm pluteus larvae survived at temperatures between 30 and 33°C, whereas 4 arm pluteus larvae survived at temperatures between 30 and 36°C for 2 h. These results suggest that the larval temperature tolerance capability of T. gratilla is stage dependent. These findings are important for understanding the life history strategy of this sea urchin in the shallow open water environment.     

The ecological energetics of Amoeba proteus (Protozoa)

The consumption, production and respiration of Amoeba proteus were measured in the laboratory for cells cultured over a range of Tetrahymena pyriformis concentrations (125–4 000 cells/0.5 ml) at 10, 15 and 20 °C. Differences were attributed to both temperature and prey availability. A series of generation energy budgets were constructed for amoebae grown under the above conditions. The biological efficiencies linking the parameters of the budget equation were calculated. Assimilation efficiencies ranged from 22–59% regardless of temperature. Net production efficiencies were high at 15 and 20 °C (65–82%) but low at 10 °C (11–49%). Gross production efficiencies were also higher at 15 and 20 °C (16–47%) than at 10 °C (4–29%). The ecological implications of this investigation are discussed.     

Germination responses to temperature responsible for the seedling emergence seasonality ofPrimula sieboldii E. Morren in its natural habitat

Temperature requirements for the breaking of seed dormancy and germination inPrimula sieboldii E. Morren and the annual surface-soil temperature regime in one of its natural habitats were investigated in order to clarify the germination responses determining the seedling emergence seasonality of the species. In a grassland nature reserve in an abandoned flood plain of the Arakawa River, natural seedling emergence of the species was shown to be restricted to mid- to late-spring before the closure of seasonal vegetational gaps, when the daily mean soil surface temperature reached about 15°C, accompanied by large daily fluctuations of about 10°C. Mature seeds collected in late June were never able to germinate at any constant temperature in the range of 8–40°C unless they had been previously subjected to moist-chilling treatment. The proportion of seeds which were released from dormancy increased with increasing duration of the moist-chilling treatment at 2°C, 70–85% of seeds becoming germinable at 16–28°C after 12 weeks of pretreatment at 2°C. The thermal time required for the germination of the thus-pretreated seed population was 905–1690 Kh with a base temperature of around 5°C. Fluctuating temperatures between 24°C and 16 or 12°C had a remarkable dormancy-breaking effect, inducing considerably quick germination in most of the seeds previously subjected to 2°C moist-chilling for 8 weeks.     

Activities of enzymes of sugar metabolism in cold-stored tubers of Solanum tuberosum

Storage of tubers of Solanum tuberosum at 10° or 2° for 15 days did not alter significantly the maximum catalytic activities of sucrose phosphate synthetase, sucrose synthetase, glucose-6-phosphate dehydrogenase, aldolase, and glyceraldehydephosphate dehydrogenase. The temperature coefficients of phosphofructokinase, glyceraldehydephosphate dehydrogenase, and pyruvate kinase from the tubers were shown to be higher between 2° and 10° than between 10° and 25°. The rate of sugar accumulation at 2° exceeded the activity of sucrose synthetase but was less than that of sucrose phosphate synthetase. It is suggested that sucrose accumulation at 2° is catalysed by sucrose phosphate synthetase, is not due to changes in the maximum catalytic activities of any of the above enzymes, but may be due, in part, to the susceptibility of key glycolytic enzymes to cold.     

Molecular characterization of four forms of phosphofructokinase purified from potato tuber

Four forms of phosphofructokinase (PFK) have been purified to apparent homogeneity from tubers of potato (Solanum tuberosum cv. Record). Each had a final specific activity of about 200 mumol.min-1.mg-1 protein. Similar forms of PFK were found in partially purified extracts from tubers and leaves of other potato cultivars and related wild species. The relative molecular masses of three forms of PFK were about 200,000 whereas that of the fourth PFK was greater than 800,000. The four forms of PFK contained different proportions of four polypeptides which had apparent relative molecular masses of 46,300, 49,500, 50,000, and 53,000. These polypeptides gave different patterns of peptide fragments after chemical and proteolytic cleavage. Western blots and immunoprecipitation studies using antibodies raised against the individual polypeptides showed that all four are associated with PFK. Thus, potato tubers contain four distinct forms of PFK that differ in their subunit composition.     

The effects of storage condition on starch metabolism and regeneration potentials of twin-scales and inflorescence stem explants of <Emphasis Type="Italic">Narcissus tazetta</Emphasis>

Summary The natural propagation rate of Narcissus is very slow. In vitro micropropagation of Narcissus is more efficient than conventional propagation for rapidly building up aseptic stocks of varieties, especially for the establishment of new cvs. and the production of pathogen-free stock material. In the present study, Narcissus tazetta cv. ‘Ziva’ bulbs were used as the source of mother plants. The bulbs were kept at 30°C in a dark chamber until the start of the experiments. Prior to explant preparation, the bulbs were subjected to a cold treatment at 15°C in the dark for 6 wk to break dormancy. Twin-scales and inflorescence stem discs were isolated from aseptic bulb parts and were used as the initial explants. The polar orientation affected the regeneration of the inflorescence stem. Storage duration at 30°C followed by cold treatment were found to affect starch levels, adenosine diphosphate glucose pyrophosphorylase (AGPase) activities, and regeneration potentials. Starch levels were reduced significantly during a 10 mo. storage period at 30°C in both twin-scales and inflorescence stem disc explants. Regeneration was followed by an efficient acclimatization system with 98–99% survival. More than 500 highly uniform young bulbs were propagated from one mother bulb in a 12 mo. period.     

Microspore embryogenesis and the development of a double haploidy protocol for cow cockle (Saponaria vaccaria)

Factors affecting microspore embryogenesis of cow cockle (Saponaria vaccaria) were evaluated including donor plant growing conditions, genotype, bud size, density, medium composition, and culture conditions. Of the two donor plant (day/night) temperature regimes evaluated (10/5°C and 20/15°C), plants grown at 20/15°C were the most embryogenic. An embryogenic frequency of greater than 350 embryos/100 buds was observed in the most embryogenic genotype, cv. ‘White Beauty’. Buds from 3–9 mm in length were evaluated for their embryogenic potential; buds that were 4–7.9 mm produced the most embryos/100 buds. Of all the media compositions evaluated, NLN medium with 15% sucrose resulted in the most embryos. Cow cockle microspores required an initial period of 32°C for 3 days for production of microspore-derived embryos (MDEs).     

Influences of Culture Temperature on the Growth, Lipid Content and Fatty Acid Composition of Aurantiochytrium sp. Strain mh0186

The growth, lipid content, and fatty acid composition of Aurantiochytrium sp. strain mh0186 at different temperatures were investigated. Strain mh0186 grew well at 15–30°C, but weakly at 10°C. The biomass at 15–30°C was significantly higher than at 10 and 35°C, and the total lipid at 15–35°C was significantly higher than that at 10°C. The amount of DHA in the total fatty acid was highest at 10°C and decreased in response to temperature increase. The content of DHA (mg/g-dry cell weight) at 15–30°C were significantly higher than those at 35°C and those at 15–25°C were significantly higher than those at 10 and 35°C. The DHA yield at 15–35°C was significantly higher than those at 10 and 35°C. Unsaturation of fatty acid was regulated by temperature and was enhanced in response to temperature decrease. The ratio of DHA to DPA varied at different temperatures.     

Characterization of proteolytic bacteria from the Aleutian deep-sea and their proteases

Six deep-sea proteolytic bacteria taken from Aleutian margin sediments were screened; one of them produced a cold-adapted neutral halophilic protease. These bacteria belong to Pseudoalteromonas spp., which were identified by the 16S rDNA sequence. Of the six proteases produced, two were neutral cold-adapted proteases that showed their optimal activity at pH 7–8 and at temperature close to 35°C, and the other four were alkaline proteases that showed their optimal activity at pH 9 and at temperature of 40–45°C. The neutral cold-adapted protease E1 showed its optimal activity at a sodium chloride concentration of 2 M, whereas the activity of the other five proteases decreased at elevated sodium chloride concentrations. Protease E1 was purified to electrophoretic homogeneity and its molecular mass was 34 kDa, as estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The molecular weight of protease E1 was determined to be 32,411 Da by mass spectrometric analysis. Phenylmethyl sulfonylfluoride (PMSF) did not inhibit the activity of this protease, whereas it was partially inhibited by ethylenediaminetetra-acetic acid sodium salt (EDTA-Na). De novo amino acid sequencing proved protease E1 to be a novel protein.     

Newly Isolated Bacillus gibsonii S-2 Capable of using Sugar Beet Pulp for Alkaline Pectinase Production

Summary A Bacillus strain capable of growing under highly alkaline conditions was isolated from a sample of alkaline soil. The isolate was a Gram-positive, spore-forming, aerobic, alkaliphilic bacterium and was designated as strain S-2. Growth of the strain was observed in the pH range of 7–12 and temperature range of 4–40 °C. Sequence analysis of the 16S rDNA of the strain revealed more than 99% homology with the strains of Bacillus gibsonii. The S-2 strain was confirmed as B. gibsonii by comparing its physiological and biochemical characteristics with the B. gibsonii DSM 8722 strain. The S-2 strain could use sugar beet pulp as the carbon source as well as the pectinase inducer to produce extracellular alkaline pectinase by solid-state fermentation. The maximum polygalacturonase yield of 3600 U/g dry sugar beet pulp was obtained at 35 °C after 48 h of incubation.     

Methods of short-term storage of cultures of the blowfly <Emphasis Type="Italic">Calliphora Vicina</Emphasis> R.-D. (Diptera,Calliphoridae)

Three methods of short-term storage of the blowfly Calliphora vicina strains are considered based on the experimental study of 21 strains originating from different parts of the species range. The colony can be preserved as diapausing adults at 6° and darkness for 2–3 months or more, depending on the geographical origin of the population. During the first five days of adult life the flies should be kept at 12° and short day on a sugar diet, after which they should be transferred into a refrigerator. During artificial hibernation the flies also require periodic sugar feeding every 20 days (3–4 h at 20°C) to maintain their vital functions. The combination of temperatures of 20–23°C and a protein diet terminates reproductive diapause, and oviposition starts in 10–17 days. The fly strain may be preserved as reproductive females at 6°C and darkness with sugar feeding. Flies also require periodic sugar feeding at 20°C (3–4 hours). In this case the flies start laying eggs 2–3 days after being transferred to 20–23°C. The preservation of diapausing larvae is a more reliable method of prolonged strain storage. In this case the flies of maternal generation are maintained at 20–23°C on sugar and protein diet. The egg rafts laid during 5–6 hours are then transferred into 12°C and short day until hatchment. The hatched larvae should be immediately placed into a refrigerator (2–3 or 5–6°C), where they feed during 1–1.5 months and enter diapause. For strain restoration, the diapausing larvae should be transferred into 20–25°C, where they pupariate in 3–5 days and the flies emerge in nearly 10 days.     

Lipid phase transition in the plasma membrane of the goat epididymal maturing spermatozoa

Microviscosity of the highly purified plasma membranes isolated from the maturing goat caput, corpus and cauda epididymal sperm, was measured using l,6-diphenyl-l,3,5-hexatriene as the lipophilic probe at varying temperatures (12–42°C). As shown by the Arrhenius plot of the data each of the maturing sperm membranes had two distinct lipid phase transitions in the temperature zones 19–25°C and 34–37°C. The low-temperature transitions for the immature caput- and mature cauda-sperm membranes were noted at 19–20°C, and 24–25°C, respectively, whereas both these membranes showed high temperature transition at 36–37°C. The maturing corpus-sperm membrane had phase transitions at 21–22°C and 35–36°C that were significantly different from those of the immature/mature sperm membranes. The data implicate significant alteration of the sperm membrane structure during epididymal maturation. The phase transition of the mature male gametes at 36–37°C may have a great impact on the subsequent events of the sperm life cycle since the mature spermatozoa that are stored in the epididymis a few degrees below the body temperature, experience higher temperature when ejaculated into the female reproductive tract.     

Irreversible changes in barley leaf chlorophyll fluorescence detected by the fluorescence temperature curve in a linear heating/cooling regime

The chlorophyll fluorescence (F) temperature curves in a linear time-temperature heating/cooling regime were used to study heat-induced irreversible F changes in primary green leaves of spring barley (Hordeum vulgare L. cv. Akcent). The leaf segments were heated in a stirred water bath at heating rates of 0.0083, 0.0166, 0.0333, and 0.0500 °C s⁻¹ from room temperature up to maximal temperature T _m and then linearly cooled to 35 °C at the same rate. The F intensity was measured by a pulse-modulated technique. The results support the existence of the two critical temperatures of irreversible F changes postulated earlier, at 45–48 and 53–55 °C. The critical temperatures are slightly dependent on the heating rate. Two types of parameters were used to characterize the irreversibility of the F changes: the coefficient of irreversibility μ defined as the ratio of F intensity at 35 °C at the starting/ending parts of the cycle and the slopes of tangents of linear parts of the F temperature curve. The dependence of μ on T _m revealed a maximum, which moved from 54 to 61 °C with the increasing heating/cooling rate v from 0.0083 to 0.0500 °C s⁻¹, showing two basic phases of the irreversible changes. The Arrhenius and Eyring approaches were applied to calculate the activation energies of the initial increase in μ. The values varied between 30 and 50 kJ mol⁻¹ and decreased slightly with the increasing heating rate.     

Degradation of RNA during the autolysis of Saccharomyces cerevisiae produces predominantly ribonucleotides

Autolytic degradation of yeast RNA occurs in many foods and beverages and can impact on the sensory quality of the product, but the resulting complex mixture of nucleotides, nucleosides and nucleobases has not been properly characterised. In this study, yeast autolysis was induced by incubating cell suspensions of Saccharomyces cerevisiae at 30–60 °C (pH 7.0), and at pH 4.0–7.0 (40 °C) for 10–14 days, and the RNA degradation products formed during the process were determined by reversed-phase HPLC. Up to 95% of cell RNA was degraded, with consequent leakage into the extracellular environment of mainly 3′-, 5′- and 2′-ribonucleotides, and lesser amounts of polynucleotides, ribonucleosides and nucleobases. The rate of RNA degradation and the composition of the breakdown products varied with temperature and pH. RNA degradation was fastest at 50 °C (pH 7.0). Autolysis at lower temperatures (30 °C and 40 °C) and at pH 5.0 and 6.0 favoured the formation of 3′-nucleotides, whereas autolysis at 40 °C and 50 °C (pH 7.0) favoured 5′- and 2′-nucleotides. The best conditions for the formation of the two flavour-enhancing nucleotides, 5′-AMP and 5′-GMP, were 50 °C (pH 7.0) and pH 4.0 (40 °C), respectively.     

An unusual cyanobacterium from saline thermal waters with relatives from unexpected habitats

Cyanobacteria that grow above seawater salinity at temperatures above 45°C have rarely been studied. Cyanobacteria of this type of thermo-halophilic extremophile were isolated from siliceous crusts at 40–45°C in a geothermal seawater lagoon in southwest Iceland. Iceland Clone 2e, a Leptolyngbya morphotype, was selected for further study. This culture grew only at 45–50°C, in medium ranging from 28 to 94 g L⁻¹ TDS, It showed 3 doublings 24 h⁻¹ under continuous illumination. This rate at 54°C was somewhat reduced, and death occurred at 58°C. A comparison of the 16S rDNA sequence with all others in the NCBI database revealed 2 related Leptolyngbya isolates from a Greenland hot spring (13–16 g L⁻¹ TDS). Three other similar sequences were from Leptolyngbya isolates from dry, endolithic habitats in Yellowstone National Park. All 6 formed a phylogenetic clade, suggesting common ancestry. These strains shared many similarities to Iceland Clone 2e with respect to temperature and salinity ranges and optima. Two endolithic Leptolyngbya isolates, grown previously at 23°C in freshwater medium, grew well at 50°C but only in saline medium. This study shows that limited genotypic similarity may reveal some salient phenotypic similarities, even when the related cyanobacteria are from vastly different and remote habitats.     

Gap-detecting mechanism in the seed germination ofMallotus japonicus (Thunb.) Muell. Arg., a common pioneer tree of secondary succession in temperate Japan

Germination responses ofMallotus japonicus (Thumb). Muell. Arg. seeds to temperature revealed a gap-detecting mechanism in the seed germination of the species. Among various constant and alternating temperatures examined in the range from 12–40°C, only very limited temperature regimes were found to be favourable for seed germination, specifically, alternating temperatures between 18–32°C and 28–40°C. A single several-hour higher-temperature (32–40°C) treatment could also induce the germination of seeds which had been imbibed for several days at a constant temperature in the range of 20–26°C, suggesting that there is a process requiring higher temperature among the overal germination processes. Seeds located at or near the surface of denuded soil would have a good chance of experiencing such a temperature change when several rainy days are followed by fine weather, while seeds beneath close vegetation would not. On the other hand, the pressence or absence of light or a simulated ‘canopy ligh’ had little effect on the germination. Therefore, it was concluded that the seeds ofM. japonicus have a ‘gapdetecting mechanism’ in the form of a higher-temperature requirement of a certain process involved in the overall germination processes.     

Thermal preference and avoidance in cladoceran <Emphasis Type="Italic">Daphnia magna</Emphasis> strauss (crustacea,cladocera) acclimated to constant temperature

The final preferable temperature (FPT) and avoidance temperature (AT) were determined in parthenogenetic females of the crustacean Daphnia magna Strauss. The animals were preliminary acclimated to constant temperature of 23.4°C followed by keeping them in a thermo-gradient device for 24 days. It was revealed that daphnia select FPT with overshoot. In the first four days, daphnia selected temperatures 0.6–1.6°C higher than the acclimation temperature and 4–7.4°C higher than FPT. Two zones of FPT are revealed: the first zone by the time of 5–13 days (17.6 ± 1.2°C); the second, by 16–24 days (20 ± 1.5°C). The dynamics of AT diapason followed the dynamics of FPT. Two zones of the AT plateau were observed: over five to 17 days (temperatures <14°C and >25.8°C were avoided) and for 21–24 days (<8.5°C and >26°C).     

Salinity and Temperature Effects on Germination for Three Salt-resistant Euhalophytes, Halostachys caspica, Kalidium foliatum and Halocnemum strobilaceum

The effects of three salinities (0, 100 and 500 mM NaCl) and four constant temperatures (10, 20, 30 and 35 °C) on seed germination of Halostachys caspica (M. B.) C. A. Mey., Kalidium foliatum (Pall.) Mop. and Halocnemum strobilaceum (Pall.) Bieb. were investigated. After seeds were treated with different concentrations of NaCl at constant temperatures of 10–35 °C for 16 days, ungerminated seeds were transferred to distilled water for 10 days to investigate the total germination; after this time, the ungerminated seeds from the 10 and 20 °C treatments were then moved to 35 °C for another 5 days to determine the final germination. The three plant species in the present experiment are salt-resistant euhalophytes growing in high saline soils in the Zhungur Basin in Xinjiang, a northwest province of China.Compared with germination under control conditions, germination percentages of all three species were not affected by 100 mM NaCl at 10–35 °C, while severely inhibited by 500 mM NaCl; germination percentages were very low at 10 °C up to 100 mM NaCl for all species; the optimum temperature for germination of H. caspica and K. foliatum was 20–30 °C, while 35 °C for H. strobilaceum, up to 100 mM NaCl; seeds did not suffer ion toxicity for all species, as evidenced by the high total germination after ungerminated seeds pretreated with 500 mM NaCl were transferred to distilled water at constant temperatures of 10–35 °C for 10 days, and the high final germination after the ungerminated seeds from the 10 and 20 °C treatments were subsequently moved to 35 °C for another 5 days; Halostachys caspica had greater sensitivity to increasing temperatures from 10 and 20 °C to 35 °C compared with the other two species.