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Design of a high-power NMR probe for low-temperature studies.

A low-temperature, high-power NMR probe head design is described which eliminates the problem of electric arc discharge commonly experienced during radiofrequency pulse cycling in a helium environment. A polychlorotrifluoroethylene (Kel-F) coil former, fitted with a solenoid coil, is heat-shrunk onto stainless-steel flanges and spot-welded inside a stainless-steel probe head assembly connected to a hollow coaxial transmission-line probe shaft. By this means, the sample coil and all high-voltage elements can effectively be isolated in a vacuum, while at the same time permitting good thermal contact between the sample and cryogenic gas. This design was used in NMR studies in the 4.6 K < or = T < or = 77 K temperature range for RF pulse durations < or = 50 ms (and longer for low RF amplitudes) and amplitudes up to approximately 60 G.

Cold Temperature↗

Uncovering molecular regulatory networks of low-temperature stress response in Trachinotus ovatus via integrated transcriptome and metabolome analyses.

Golden pompano (Trachinotus ovatus) is one of the most economically important marine fish species in China. It is susceptible to low-temperature stress, which significantly challenges its production and supply. Nevertheless, study on the regulatory mechanisms underlying low-temperature stress responses in golden pompano remains limited. Here, we firstly performed a time-series transcriptome analysis to reconstruct dynamic response patterns under low-temperature stress in golden pompano. Transcriptome profiling identified common differentially expressed genes (DEGs), including fos, hlf, and hmgb1, as well as condition-specific DEGs across distinct low-temperature stress groups. Based on cluster analysis, all DEGs were classified into five distinct expression patterns, reflecting diversified regulation of expression in golden pompano during low-temperature stress. Furthermore, condition-specific regulatory modules were explored via weighted gene co-expression network analysis (WGCNA), highlighting that the two module hub genes, serbf2 and lipc, might respond to low-temperature stress by regulating the lipid catabolic process. Subsequently, untargeted metabolomic analysis revealed that glycerophospholipid metabolism was a significantly enriched common pathway, highlighting its crucial role in mediating the response to low-temperature stress. Finally, by integrating transcriptomic and metabolomic analyses, a gene-metabolite interaction network associated with glycerophospholipid metabolism under low-temperature stress was established. These findings underscore the significance of multiple candidate genes and glycerophospholipid metabolism in golden pompano's response to low-temperature stress, thereby laying a solid molecular foundation for the development of low-temperature-tolerant fish strains.

Animals↗

The effect of low-temperature-induced DNA supercoiling on the expression of the desaturase genes in synechocystis.

Temperature-dependent changes in genomic DNA supercoiling might play an important role in temperature perception and responsive gene regulation. In the cyanobacterium Synechocystis, low temperatures induce the expression of the genes for fatty acid desaturases that introduce double bonds into acyl chains of lipid-bound fatty acids, thus regulating the membrane fluidity. I studied the effects of low temperature on supercoiling of the genomic DNA region that contains the regulatory elements of the desB gene for the omega3 desaturase, which is strongly induced by cold. Upon decrease in temperature, the degree of DNA supercoiling increased in this region. Novobiocin, an inhibitor of the DNA gyrase, prevented low-temperature-induced changes in DNA supercoiling and affected the expression of several desaturase genes with the most effect on desB. Decreasing in temperature induces three genes of FA desaturases encoding delta12, delta6 and omega3 desaturases in Synechocystis cells. Novobiocin inhibited completely low-temperature-induced transcription of desB, accumulation of the corresponding protein, and the formation of the omega3 unsaturated fatty acids. In the presence of novobiocin, the novobiocin-resistant mutant cells of Synechocystis responded to the low-temperature treatment in the same way as the wild-type cells in the absence of the antibiotics. Thus, temperature-induced changes in DNA supercoiling might form an essential part of a global regulation pathway leading to low-temperature acclimation in this mesophilic cyanobacterium.

Amino Acid Sequence↗

C(4) Pathway Photosynthesis at Low Temperature in Cold-tolerant Atriplex Species.

Two species of Atriplex were grown under low temperature (8 C day/6 C night) and high temperature (28 C day/20 C night) regimes. The photosynthetic capacity of these plants was studied as a function of temperature in a leaf gas exchange cuvette. Both species showed substantial photosynthetic capacity between 4 and 10 C and this was not enhanced by growth at low temperatures but rather, was somewhat greater in plants grown at higher temperature. Photosynthetic capacity of low temperature-grown plants at high temperature was greater in Atriplex confertifolia (Torr. and Frem.) S. Watts., a native of cool deserts, than in Atriplex vesicaria (Hew. ex. Benth.) from warmer desert areas. Leaves of both species were also subjected to (14)CO(2) pulse-chase and steady-state feeding experiments under controlled temperature conditions. These experiments revealed that the kinetics of carbon assimilation through the intermediates of the C(4) pathway is not substantially disrupted at low temperature in either species. There was, however, a substantial interchange of label between aspartate and malate at low temperature which was not evident at high temperature. There was also an increase in the pool sizes of the C(4) acids involved in photosynthesis of A. confertifolia. Speculation as to the explanation of these changes and their possible significance in promoting low temperature C(4) photosynthesis in these plants is presented.

Journal Article↗

Low-temperature induction of Myxococcus xanthus developmental gene expression in wild-type and csgA suppressor cells.

The csgA gene encodes an extracellular protein that plays an essential role in the regulation of fruiting-body formation and sporulation of Myxococcus xanthus. The csgA suppressor allele soc-500 (formerly referred to as csp-500) was selected based on its ability to restore sporulation to csgA cells under developmental conditions at 32 degrees C. The soc-500 allele was subsequently found to induce sporulation of csgA+ or csgA cells simply by shifting the temperature of vegetatively growing cells to 15 degrees C. Low-temperature-induced sporulation of soc-500 strains occurred in the absence of two requirements for fruiting-body sporulation: low nutrient levels and a high temperature. Low temperature alone caused the expression of many developmentally regulated genes but did not support the development of wild-type cells. The soc-500 allele appears to activate genes involved with sensing nutritional stress. At low temperature on a nutritionally rich medium, soc-500 induced expression of the tps gene which is normally expressed following nutritional shiftdown. The soc-500 allele was cloned and integrated into the wild-type chromosome by site-specific recombination. It was dominant over the wild-type allele in merodiploids and is contained on a 3-kbp DraI-ClaI restriction fragment. The soc-500 transcriptional unit spans a 300-bp PstI-PstI restriction fragment, since deletion of the PstI restriction fragment inhibits both csgA suppression and low-temperature induction. These results suggest that the soc-500 mutation lies in a gene that is involved in nutrient sensing.

Alleles↗

Mycorrhiza does not alter low temperature impact on Gnaphalium norvegicum.

Extreme arctic-alpine vegetation has relatively low affinity to form mycorrhizal symbiosis. We asked whether the mycorrhizal growth benefit for the host plant is lower at low temperatures. We investigated the role of two root-associated fungi and temperature in growth, carbon-nitrogen relations and germination of an arctic-alpine herb. Seeds of Gnaphalium norvegicum were germinated at 8 degrees or 15 degrees C with or without arbuscular mycorrhizal (AM, Glomus claroideum) and dark septate endophytic (DSE, Phialocephala fortinii) inocula in a climate chamber. We found that germination percentage, shoot and root biomass, shoot N% and root AM colonization were lower at 8 degrees C than at 15 degrees C. P. fortinii inoculation had a positive impact on germination at both temperatures, whereas G. claroideum produced no effect. N% was lower in AM plants at both temperatures. Plant biomass and shoot N content were higher in AM plants than in control plants at 15 degrees C, but not at 8 degrees C. DSE inoculation tended also to have positive effects on plant biomass and N content at 15 degrees C. At 15 degrees C, rate of photosynthesis, photosynthetic nutrient use efficiency and specific leaf area were positively affected by G. claroideum, which suggests that G. claroideum formed a carbon sink and possibly enhanced the seedling water economy. The positive effects of P. fortinii were probably due to its saprotrophic function in the substrate because it did not colonize the roots. These results suggest that the effects of AM and DSE on plant growth are affected by temperature and that the mycorrhizal benefit for the host plant was lower at the lower temperature. Low saprotrophic activity and decreased mycorrhiza-mediated nutrient acquisition may thus constrain plant nutrient acquisition in cold environments. Decreased mycorrhizal benefit may be related to the comparatively low mycotrophy of cold environment vegetation.

Analysis of Variance↗

Low-temperature activation conditions for PAMAM dendrimer templated Pt nanoparticles.

Surface immobilized polyamidoamine (PAMAM) dendrimer templated Pt nanoparticles were employed as precursors to heterogeneous catalysts. CO oxidation catalysis and in situ infrared spectroscopy were used to evaluate conditions for dendrimer removal. Infrared spectroscopy showed that PAMAM dendrimer amide bonds begin decomposing at temperatures as low as 75 degrees C. Although the amide stretches are completely removed after 3 h of oxidation at 300 degrees C, 16 h were required to reach maximum catalytic activity. Further treatment under oxidizing or reducing atmospheres did not cause substantial changes in activity. Infrared spectroscopy of the activated materials indicated that organic residues, probably surface carboxylates, are formed during oxidation. These surface species passivate the Pt NPs, and their removal was required to fully activate the catalyst. Substantially less forcing activation conditions were possible by employing a CO/O(2)/He oxidation treatment. At appropriate temperatures, CO acts as a protecting group for the Pt surface, helping to prevent fouling of the nanoparticle by organic residues. CO oxidation catalysis and infrared spectroscopy of adsorbed CO indicated that the low temperature activation treatment yielded supported nanoparticles that were substantially similar to those prepared with more forcing conditions.

Journal Article↗

Differential impact of low temperature on fatty acid unsaturation and lipoxygenase activity in figleaf gourd and cucumber roots.

Previous studies show that low temperature strongly induces suberin layers in the roots of chilling-sensitive cucumber plants, while in contrast, low temperature produces a much weaker induction of suberin layers in the roots of the chilling-tolerant figleaf gourd [S.H. Lee, G.C. Chung, S. Steudle, Gating of aquaporins by low temperature in roots of chilling-sensitive cucumber and -tolerant figleaf gourd, J. Exp. Bot. 56 (2005) 985-995; S.H. Lee, G.C. Chung, E. Steudle, Low temperature and mechanical stresses differently gate aquaporins of root cortical cells of chilling-sensitive cucumber and figleaf gourd, Plant Cell Environ. (2005) in press; S.J. Ahn, Y.J. Im, G.C. Chung, B.H. Cho, S.R. Suh, Physiological responses of grafted-cucumber leaves and rootstock roots affected by low root temperature, Scientia Hort. 81 (1999) 397-408]. Here, the effect of low temperature on fatty acid unsaturation and lipoxygenase activity was examined in cucumber and figleaf gourd. The double bond index demonstrated that membrane lipid unsaturation shows hyperbolic saturation curve in figleaf gourd roots while a biphasic response in cucumber roots to low temperature. In figleaf gourd, the hyperbolic response in the double bond index was primarily due to accumulation of linolenic acid. Chilling stress also significantly induced lipoxygenase activity in figleaf gourd roots. These results suggest that the degree of unsaturation of root plasma membrane lipids correlates positively with chilling-tolerance. Therefore, studies that compare the effects of chilling on cucumber and figleaf gourd may provide broad insight into stress response mechanisms in chilling-sensitive and chilling-tolerant plants. Furthermore, these studies may provide important information regarding the relationship between lipid unsaturation and lipoxygenase function/activity, and between lipoxygenase activity and water channeling during the response to chilling stress. The possible roles of these processes in chilling tolerance are discussed.

Cell Membrane↗

Effect of low temperature on radiation-induced genetic damage in Drosophila melanogaster: response of motile sperm and late spermatids.

The response of fully mature motile sperm and late spermatids when challenged with X-radiation at 0 degrees C has been studied in sex-linked recessive lethals, II-III translocations and dominant lethality experiments. At 0 degrees C a significant increase in both mutagenic and clastogenic damage was detected compared to that obtained at 24 degrees C. Furthermore, the results of experiments performed with different postirradiation temperatures demonstrate that the low temperature during irradiation was the sole factor responsible for the observed increase. In the recessive lethal and translocation tests the response of late spermatids was higher than that shown by motile spermatozoa. As a whole, the results, which are rather similar to data reported on the effect of irradiation in oxygen of the same cell stages, suggest that the low temperature acted as a dose-modifying factor.

Animals↗

[Effect of the delta-sleep peptide on erythrocyte membrane function during low-temperature exposure].

3-day effect of low temperature increases the amount of extraerythrocyte hemoglobin by 64% in the blood serum, the activity of glucose-6-phosphatedehydrogenase increasing by 231%. Administration of 6.12 or 18 micrograms/100 g body weight of the delta-sleep peptide (DSIP) induces on change in amount of extraerythrocyte hemoglobin in the blood serum of intact rats. 6 micrograms/100 g DSIP increases the activity of glucose-6-phosphatedehydrogenase by 251.6%; 12 micrograms/100 g-- by 165.6%; 18 micrograms/100 g--by 90.6%. The normalizing effect on the level of extraerythrocyte hemoglobin and activity of glucose-6-phosphatedehydrogenase in low ambient temperature occurs at the dose of DSIP 12 micrograms/100 g body weight. The DSIP stabilizes the erythrocyte membranes in low temperature.

Animals↗

Identification of promoter elements in a low-temperature-responsive gene (blt4.9) from barley (Hordeum vulgare L.).

The blt4 barley gene family encodes non-specific lipid transfer proteins and has been shown, by in situ localisation, to be expressed in the epidermal cells of leaves. The transcriptionally controlled, low-temperature-responsive member of this gene family, blt4.9, is predominantly expressed in shoot meristems. The promoter region (1938 bp) of blt4.9 contains sequence motifs which have been implicated in responses to low temperature, abscisic acid and other environmental factors. Deletion analysis showed that a 42 bp sequence proximal to, but not including, the CAAT and TATA boxes, confers enhanced low-temperature response to a reporter gene in a barley shoot explant transient expression system. Other promoter regions were shown to contain negative and positive regulatory regions. Electrophoretic mobility shift analysis (EMSA) was used with nuclear proteins from either low-temperature- or control-temperature-treated plants to further investigate the blt4.9 promoter. Synthetic oligonucleotides were used to identify a hexanucleotide, CCGAAA, within the 42 bp, low-temperature-responsive promoter region, as the binding site of a low-mobility nuclear protein complex. This complex was present in nuclear extracts from both low-temperature-treated and control plants and was the only complex formed within this region. Mutation of the CCGAAA motif within the low-temperature-responsive 42 bp promoter sequence reduced low-temperature responsiveness to basal levels. A related upstream element, CCGAC, known to be a low-temperature-responsive element in other plants, did not bind to nuclear proteins in this study. It is proposed that the hexanucleotide CCGAAA, at -195 from the first ATG, is involved in the low-temperature response of blt4.9 in barley.

Base Sequence↗

Diamond formation by reduction of carbon dioxide at low temperatures.

This Communication reports a low-temperature diamond synthesis technique, in which diamonds (10-250 mum) can form at a temperature as low as 440 degrees C by reduction of dense CO2 with metallic Na. The X-ray diffraction pattern of a powder sample shows three reflection peaks, indexed with 111, 220, and 311, corresponding unambiguously to cubic diamond. The Raman spectrum of the product exhibits an intense first-order peak at 1332 cm-1, which is the characteristic signature of the cubic diamond, indicating the formation of well-crystallized diamond. Carbon dioxide is a nontoxic low-energy molecule, abundant on earth. This novel reduction method could allow studies of large-size diamond growth using CO2 as the carbon source.

Journal Article↗

A maize DNA-binding factor with a bZIP motif is induced by low temperature.

We have isolated a low temperature-induced maize gene, mlip15, via cross hybridization using rice lip19. The longest cDNA isolated comprised 1179 bp and coded for a 135 amino acid bZIP (basic region/leucine zipper) protein. The gene showed 61.4% and 68.9% identity with the rice gene at the DNA and amino acid sequence levels, respectively, and is distinct from other maize genes that code for bZIP proteins. The level of mlip15 transcript was positively regulated by low temperature in the same way as the lip19 transcript. The levels of the transcript were also strongly increased by salt stress and exogenous abscisic acid, and slightly increased by anaerobiosis, but were not affected by heat shock and drought. The mLIP15 protein and truncated derivatives, produced in rabbit reticulocyte lysates or in an Escherichia coli expression system, were able to bind to a fragment of the wheat histone H3 gene promoter. This binding was diminished by addition of a molar excess of the hexamer sequence 5'-ACGTCA-3' found in the promoter and of the G-box-like sequence, but not by the addition of the ocs sequence or a mutated hexamer sequence. The factor bound to a promoter region of the maize Adh1 gene, expression of which is also induced by low temperature. These results lead to the conclusion that mlip15 is a strong candidate for a low temperature-induced transcription factor in maize.

Amino Acid Sequence↗

Two transcription factors, DREB1 and DREB2, with an EREBP/AP2 DNA binding domain separate two cellular signal transduction pathways in drought- and low-temperature-responsive gene expression, respectively, in Arabidopsis.

Plant growth is greatly affected by drought and low temperature. Expression of a number of genes is induced by both drought and low temperature, although these stresses are quite different. Previous experiments have established that a cis-acting element named DRE (for dehydration-responsive element) plays an important role in both dehydration- and low-temperature-induced gene expression in Arabidopsis. Two cDNA clones that encode DRE binding proteins, DREB1A and DREB2A, were isolated by using the yeast one-hybrid screening technique. The two cDNA libraries were prepared from dehydrated and cold-treated rosette plants, respectively. The deduced amino acid sequences of DREB1A and DREB2A showed no significant sequence similarity, except in the conserved DNA binding domains found in the EREBP and APETALA2 proteins that function in ethylene-responsive expression and floral morphogenesis, respectively. Both the DREB1A and DREB2A proteins specifically bound to the DRE sequence in vitro and activated the transcription of the b-glucuronidase reporter gene driven by the DRE sequence in Arabidopsis leaf protoplasts. Expression of the DREB1A gene and its two homologs was induced by low-temperature stress, whereas expression of the DREB2A gene and its single homolog was induced by dehydration. Overexpression of the DREB1A cDNA in transgenic Arabidopsis plants not only induced strong expression of the target genes under unstressed conditions but also caused dwarfed phenotypes in the transgenic plants. These transgenic plants also revealed freezing and dehydration tolerance. In contrast, overexpression of the DREB2A cDNA induced weak expression of the target genes under unstressed conditions and caused growth retardation of the transgenic plants. These results indicate that two independent families of DREB proteins, DREB1 and DREB2, function as trans-acting factors in two separate signal transduction pathways under low-temperature and dehydration conditions, respectively.

Adaptation, Physiological↗