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Cold acclimation induced genes of trifoliate orange (Poncirus trifoliata).

Commercial citrus varieties are sensitive to low temperature. Poncirus trifoliata is a close relative of Citrus species and has been widely used as a cold-hardy rootstock for citrus production in low-temperature environments. mRNA differential display-reverse transcription (DDRT)-PCR and quantitative relative-RT-PCR were used to study gene expression of P. trifoliata under a gradual cold-acclimation temperature regime. Eight up-regulated cDNA fragments were isolated and sequenced. These fragments showed high similarities at the amino acid level to the following genes with known functions: betaine/proline transporter, water channel protein, aldo-keto reductase, early light-induced protein, nitrate transporter, tetratricopeptide-repeat protein, F-box protein, and ribosomal protein L15. These cold-acclimation up-regulated genes in P. trifoliata are also regulated by osmotic and photo-oxidative signals in other plants.

Acclimatization↗

Cold acclimation and cryoprotectants in a freeze-tolerant Antarctic nematode, Panagrolaimus davidi.

Panagrolaimus davidi is a freeze-tolerant Antarctic nematode which survives extensive intracellular freezing. This paper describes the development of culture techniques which provide clean samples, with a high degree of freeze tolerance and in sufficient quantities for the analysis of potential cryoprotectants. Cultures grown at 20 degrees C survived a short-term freezing stress but survival declined with the time spent frozen. Acclimation of cultures at 5 degrees C enhanced the long-term survival of freezing. Starvation, however, reduced the nematode's ability to survive short-term freezing. The principal cryoprotectants detected by gas chromatography were trehalose and glycerol. The levels of trehalose, but not those of glycerol, increased significantly after acclimation. Trehalose may stabilise membranes and protect them against the dehydrating effects of the osmotic stresses resulting from freeze concentration effects but other factors, such as recrystallisation inhibition, may be involved in long-term survival.

Acclimatization↗

Selected ambient temperatures of rats acclimated to heat given on various schedules.

We investigated the behavioral thermoregulation of heat-acclimated rats by measuring their selected ambient temperatures (Ts). Rats kept in a light:dark cycle of 12:12 h were subjected to one of four different heat exposure regimes for 10 consecutive days; a constant ambient temperature (Ta) of 32 degrees C (CH), a Ta of 32 degrees C for 5 h daily in the latter half of the dark phase (IHF), a Ta of 32 degrees C for 5 h daily at a random time of day (IHR), or a constant Ta of 24 degrees C (control). After the heat exposure schedule, the rats were placed in a thermal gradient and their intra-abdominal temperature (Tab), Ts and spontaneous activity were measured for 3 days. There were clear day-night variations of Tab and Ts in all groups. The levels of Tab and Ts of the CH rats were significantly higher than those of the IHF, IHR and control rats. The Tab and activity levels of the IHF rats were significantly lower than those of the IHR and control rats only in the latter half of the dark phase. The Ts values of the IHF rats did not differ from those of the IHR and control rats. These results suggest that, after rats were constantly subjected to heat, heat-seeking behavior was induced so that the core temperature was maintained at a high level. However, when rats were acclimated to heat given for several hours at a fixed time daily, core temperature was lowered during the same time period of previous heat exposure in association with a depression of thermogenic behavior.

Abdomen↗

Transcriptional regulation of genes encoding subunits of photosystem I during acclimation to high-light conditions in Synechocystis sp. PCC 6803.

Cyanobacteria, algae and plants reduce photosystem contents and modulate photosystem stoichiometry upon acclimation to high-light (HL) conditions to avoid the damage due to excess light energy. In order to elucidate the molecular mechanism of HL acclimation, mRNA levels of genes encoding subunits of photosystems were examined in response to change of photon flux density in the wild-type strain of Synechocystis sp. PCC 6803. Transcript levels and promoter activities of photosystem I (PSI) genes rapidly decreased upon the shift to HL to less than 10% of the initial level within 1 h, whereas responses of photosystem II (PSII) transcript levels were not coordinated. The prompt change in promoter activities of PSI genes, but not PSII genes, seems important for Synechocystis cells to regulate their photosystem contents in response to changes in photon flux density. The mRNA stabilities of PSI genes also decreased during HL incubation. The down-regulation of PSI transcripts under HL did not depend on de novo protein synthesis, contrasting with the requirement of newly synthesized protein factor(s) for the accumulation of PSI transcripts under low light. When gene expression profiles of wild-type cells and a pmgA-disrupted mutant that cannot modulate photosystem stoichiometry under HL were compared, a large increase in the psaAB and psaA transcript levels was observed in the mutant under prolonged exposure to HL for 6 h. Repression of the promoter activity of the psaA gene by a pmgA-dependent mechanism was shown to be essential for the adjustment of photosystem stoichiometry under HL conditions.

Acclimatization↗

Cold acclimation in bryophytes: low-temperature-induced freezing tolerance in Physcomitrella patens is associated with increases in expression levels of stress-related genes but not with increase in level of endogenous abscisic acid.

Bryophyte species growing in areas in which temperatures fall below zero in winter are likely to have tolerance to freezing stress. It is well established in higher plants that freezing tolerance is acquired by exposure to non-freezing low temperatures, accompanied by expression of various genes and increases in levels of the stress hormone abscisic acid (ABA). However, little is known about the physiological changes induced by cold acclimation in non-vascular plants such as bryophytes. We examined the effects of low temperatures on protonema cells of the moss Physcomitrella patens (Hedw.) Bruch and Schimp. The freezing tolerance of protonema cells was clearly increased by incubation at low temperatures ranging from 10 degrees C to 0 degrees C, with maximum tolerance achieved by incubation at 0 degrees C for several days. The enhancement of freezing tolerance by low temperatures occurred in both light and dark conditions and was accompanied by accumulation of several transcripts for late-embryogenesis-abundant (LEA) proteins and boiling-soluble proteins. By de-acclimation, low-temperature-induced expression of these transcripts and proteins, as well as the freezing tolerance, was reduced. Interestingly, endogenous levels of ABA in tissues or that secreted into the culture medium were not specifically increased by low-temperature treatment. Furthermore, removal of ABA from the medium by addition of activated charcoal did not affect low-temperature-induced freezing tolerance of the protonema cells. Our results provide evidence that bryophytes have an ABA-independent cold-signaling pathway leading to expression of stress-related genes and resultant acquisition of freezing tolerance.

Abscisic Acid↗

Dehydrins in cold-acclimated apices of birch (Betula pubescens ehrh. ): production, localization and potential role in rescuing enzyme function during dehydration

Dehydrins accumulate in various plant tissues during dehydration. Their physiological role is not well understood, but it is commonly assumed that they assist cells in tolerating dehydration. Since in perennials the ability of the shoot apex to withstand dehydration is pivotal for survival through winter, we investigated if and how dehydrins may be involved. A first step in assessing such a role is the identification of their subcellular location. We therefore mapped the location of dehydrin homologues, abscisic acid-responsive (RAB 16-like) polypeptides, in the apex of birch (Betula pubescens Ehrh.). In non-cold-acclimated plants a single low-abundant RAB 16-member (a 33-kDa polypeptide) was produced, and localized in the cytoplasm only. During cold acclimation two additional members were produced (24 and 30 kDa) and accumulated in nuclei, storage protein bodies and starch-rich amyloplasts. Western blots of proteins isolated from purified starch granules and from protein bodies revealed the presence of the 24-kDa dehydrin. Since starch and protein reserves are gradually consumed during winter, serving cell maintenance, starch- and protein-degrading enzymes must remain locally active. We therefore investigated the hypothesis that dehydrins might create local pools of water in otherwise dehydrated cells, thereby maintaining enzyme function. In agreement with our hypothesis, enzyme assays showed that under conditions of low water activity a partially purified dehydrin fraction was able to improve the activity of alpha-amylase (EC 3.2.1.1.) relative to fractions from which dehydrin was removed by immunoprecipitation. The results confirm the general belief that dehydrins serve desiccation tolerance, and suggest that a major function is to rescue the metabolic processes that are required for survival and re-growth.

Journal Article↗

Seasonal changes in temperature and light drive acclimation of photosynthetic physiology and macromolecular content in Lobaria pulmonaria.

Lobaria pulmonaria (L.) Hoffm. is an epiphytic lichen common to temperate deciduous forests where it copes with large changes in temperature and light levels through repeated annual cycles. Samples of L. pulmonaria were taken from a deciduous forest in southeastern Canada at 35-day intervals from February 1999 to February 2000 and also from a rare population in an evergreen forest in March and August 1999. At field-ambient temperatures and light levels, the realised photosystem II (PSII) electron transport was low both in the summer and winter, with transient peaks in the spring and autumn. In contrast, the seasonal pattern of potential electron transport measured at a fixed 20 degrees C peaked in winter, showing the importance of temperature in driving photosynthesis to low levels in the winter despite an acclimation of electron-transport potential to exploit the high ambient light. Realised gross CO2 uptake was correlated with PSII electron transport at mechanistically plausible rates at all sampling sites in the summer but not in the winter, indicating electron diversion away from CO2 fixation in the winter. Chlorophyll content was highest in the dark summer months. The amount of ribulose-1,5-bisphosphate carboxylase-oxygenase (RuBisCO) large subunit (LSU) was highest in spring. Changes in the level of this hyperabundant protein and in the activity of PSII maintained a relatively constant rate of maximum CO2 uptake per RuBisCO LSU from April through November, despite great changes in the seasonal light and temperature. L. pulmonaria acclimates between light and temperature stress in the winter months to light-limitation in the dark summer months. Transition intervals in the spring and autumn, with warm, bright and wet conditions, are likely the most amenable times for growth.

Acclimatization↗

Leaf anatomy and light acclimation in woody seedlings after gap formation in a cool-temperate deciduous forest.

The photosynthetic light acclimation of fully expanded leaves of tree seedlings in response to gap formation was studied with respect to anatomical and photosynthetic characteristics in a natural cool-temperate deciduous forest. Eight woody species of different functional groups were used; two species each from mid-successional canopy species (Kalopanax pictus and Magnolia obovata), from late-successional canopy species (Quercus crispula and Acer mono), from sub-canopy species (Acer japonicum and Fraxinus lanuginosa) and from vine species (Schizophragma hydrangeoides and Hydrangea petiolaris). The light-saturated rate of photosynthesis (Pmax) increased significantly after gap formation in six species other than vine species. Shade leaves of K. pictus, M. obovata and Q. crispula had vacant spaces along cell walls in mesophyll cells, where chloroplasts were absent. The vacant space was filled after the gap formation by increased chloroplast volume, which in turn increased Pmax. In two Acer species, an increase in the area of mesophyll cells facing the intercellular space enabled the leaves to increase Pmax after maturation. The two vine species did not significantly change their anatomical traits. Although the response and the mechanism of acclimation to light improvement varied from species to species, the increase in the area of chloroplast surface facing the intercellular space per unit leaf area accounted for most of the increase in Pmax, demonstrating the importance of leaf anatomy in increasing Pmax.

Acclimatization↗

Decolorization kinetics of the azo dye reactive red 2 under methanogenic conditions: effect of long-term culture acclimation.

The biological decolorization of the textile azo dye Reactive Red 2 was investigated using a mixed, mesophilic methanogenic culture, which was developed with mixed liquor obtained from a mesophilic, municipal anaerobic digester and enriched by feeding a mixture of dextrin/peptone as well as media containing salts, trace metals and vitamins. Batch decolorization assays were conducted with the unacclimated methanogenic culture and dye decolorization kinetics were determined as a function of initial dye, biomass, and carbon source concentrations. Dye decolorization was inhibited at initial dye concentrations higher than 100 mg l(-1) and decolorization kinetics were described based on the Haldane model. The effect of long-term culture exposure to the reactive dye on decolorization kinetics, culture acclimation, as well as possible dye mineralization was tested using two reactors fed weekly for two years with an initial dye concentration of 300 mg l(-1) and a mixture of dextrin/peptone. The maximum dye decolorization rate after a 2-year acclimation at an initial dye concentration of 300 mg l(-1) was more than 10-fold higher as compared to that obtained with the unacclimated culture. Aniline and the o-aminohydroxynaphthalene derivative resulting from the reductive azo bond cleavage of the dye were detected, but further transformation(s) leading to dye mineralization were not observed. Reactive Red 2 did not serve as the carbon and energy source for the mixed culture, and dye decolorization was sustained by the continuous addition of dextrin and peptone. Thus, biological decolorization of reactive azo dyes is feasible under conditions of low redox potential created and maintained in overall methanogenic systems, but supply of a biodegradable carbon source is necessary.

Anaerobiosis↗

Degradation of mono-fluorophenols by an acclimated activated sludge.

Acclimated activated sludge was examined for its ability to degrade mono-fluorophenols as the sole carbon source in aerobic batch cultures. The acclimated activated sludge degraded fluorophenol efficiently. It degraded 100 mg/l 3-fluoropheno and 4-fluorophenol in 16 h with, respectively, 99.85% and 99.91% fluoride anion release and it degraded 50 mg/l 2-fluorophenol in 15 h with 99.26% fluoride anion release. The aerobic biodegradability of the mono-fluorophenols decreased in the order: 4-fluorophenol > 3-fluorophenol > 2-fluorophenol, resulting mainly from a different octanol/water partition coefficient and different steric parameter of the fluorophenols. The mechanism study revealed that the initial step in the aerobic biodegradation of mono-fluorophenols by the activated sludge was their transformation to fluorocatechol. Following transformation of the fluorophenol to fluorocatechol, ring cleavage by catechol 1, 2-dioxygenases proceeded via an ortho-cleavage pathway, then defluorination occurred.

Aerobiosis↗

State transitions revisited-a buffering system for dynamic low light acclimation of Arabidopsis.

The mobile part of the light-harvesting chlorophyll (chl) a/b protein complex (LHCII), composed of the Lhcb1 and Lhcb2 proteins, is the basic unit of chloroplast state transitions--the short term tuning system in balancing the excitation energy between Photosystem (PS) II and PSI. State transitions are catalysed by the thylakoid associated STN7 kinase, and we show here that besides the phosphorylation of the Lhcb1 and Lhcb2 proteins, also the phosphorylation of Lhcb4.2 (CP29) is under the control of the STN7 kinase. Upon growth of Arabidopsis WT and stn7 mutant plants under low and moderate light conditions, the WT plants favoured state 2 whereas stn7 was locked in state 1. The lack of the STN7 kinase and state transitions in stn7 also modified the thylakoid protein contents upon long-term low light acclimation resulting, for example, in low Lhcb1 and in elevated Lhca1 and Lhca2 protein amounts as compared to WT. Adjustments of thylakoid protein contents probably occurred at post-transcriptional level since the DNA microarray experiments from each growth condition did not reveal any significant differences between stn7 and WT transcriptomes. The resulting high Lhcb2/Lhcb1 ratio in stn7 upon growth at low light was accompanied by lower capacity for NPQ than in WT. On the contrary, higher amounts of PsbS in stn7 under moderate and high light growth conditions resulted in higher NPQ compared to WT and consequently also in a protection of PSII against photoinhibition. STN7 kinase and the state transitions are suggested to have a physiological significance for dynamic acclimation to low but fluctuating growth light conditions. They are shown to function as a buffering system upon short high light illumination peaks by shifting the thylakoids from state 2 to state 1 and thereby down regulating the induction of stress-responsive genes, a likely result from transient over-reduction of PSI acceptors.

Adaptation, Physiological↗

Insights into the acclimation of Chlamydomonas reinhardtii to sulfur deprivation.

During sulfur deprivation, the photosynthetic green alga Chlamydomonas reinhardtii develops a high-affinity sulfate uptake system and increases the expression of genes encoding proteins involved in sulfur assimilation. Although two regulatory elements, SAC1 and SAC3, have been shown to be required for normal acclimation of C. reinhardtii to sulfur deprivation, a number of other regulatory elements appear to also be involved. The molecular mechanisms by which these regulatory elements function are largely unknown. This manuscript presents our current knowledge of sulfur deprivation responses and the regulation of these responses in C. reinhardtii. In addition, we present preliminary results of a sub-saturation screen for novel sulfur acclimation mutants of C. reinhardtii. A speculative model, incorporating the activities of established regulatory elements with putative novel components of the signal transduction pathway(s) is discussed.

Acclimatization↗

Cold acclimation or grapeseed oil feeding affects phospholipid composition and mitochondrial function in duckling skeletal muscle.

The phospholipid fatty acid (FA) composition and functional properties of skeletal muscle and liver mitochondria were examined in cold-acclimated (CA, 4 degrees C) ducklings. Phospholipid FA of isolated muscle mitochondria from CA birds were longer and more unsaturated than those from thermoneutral (TN, 25 degrees C) reared ducklings. The rise in long-chain and polyunsaturated FA (PUFA, mainly 20:4n-6) was associated with a higher State 4 respiration rate and a lower respiratory control ratio (RCR). Hepatic mitochondria, by contrast, were much less affected by cold acclimation. The cold-induced changes in phospholipid FA profile and functional properties of muscle mitochondria were reproduced by giving TN ducklings a diet enriched in grapeseed oil (GO, rich in n-6 FA), suggesting a causal relationship between the membrane structure and mitochondrial functional parameters. However, hepatic mitochondria from ducklings fed the GO diet also showed an enrichment in long-chain PUFA but opposite changes in their biochemical characteristics (lower State 4, higher RCR). It is suggested that the differential modulation of mitochondrial functional properties by membrane lipid composition between skeletal muscle and liver may depend on muscle-specific factors possibly interacting with long-chain PUFA and affecting the proton leakiness of mitochondrial membranes.

Animals↗

Correlation between fluidity and fatty acid composition of phospholipid species in Tetrahymena pyriformis during temperature acclimation.

The correlation between the fluidity of phospholipids and their fatty acid composition was studied by spin label technique and gas-liquid chromatography for three major phospholipid species in Tetrahymena pyriformis during temperature acclimation. The fluidity of 2-aminoethylphosphonolipid increased within the first 10 h of the cold-acclimation when the content of gamma-linolenic acid in 2-aminoethylphosphonolipid was highest, and it then decreased up to 24 h. On the other hand, the fluidities of phosphatidylethanolamine and phosphatidylcholine showed a gradual decrease up to 24 h after the temperature shift, although gamma-linolenic acid contents were highest at 10 h after the temperature shift. Thus the fluidity changes of these two phospholipids were interpreted as resulting from the altered content of other fatty acids in addition to gamma-linolenic acid, since the gamma-linolenic acid content was smaller than that of 2-aminoethylphosphonolipid. The results suggest that the content of gamma-linolenic acid in 2-aminoethylphosphonolipid plays a role in regulating the thermal adaptation process.

Acclimatization↗

Rapid membrane response during low-temperature acclimation. Correlation of early changes in the physical properties and lipid composition of Tetrahymena microsomal membranes.

When Tetrahymena pyriformis cells grown at 39 degrees C were chilled to 15 degrees C, a rapid desaturation of microsomal phospholipid-bound fatty acids was observed. A concurrent but even more rapid change in the physical properties of the microsomal lipids was detected by steady-state fluorescence polarization measurements of the probe 1,6-diphenyl-1,3,5-hexatriene in lipid multilamellar vesicles. Whereas polarization vs. temperature plots of lipids from 39 degrees C-grown cells showed discrete break points (abrupt slope changes thought to indicate altered phase separation rates) at characteristic temperatures, plots made using lipids from equivalent cells chilled to 15 degrees C for 15 or 20 min lacked such clearly defined break points. The sharp break points reappeared in plots of microsomal lipids from cells maintained at 15 degrees C for 30 min or longer, but in these curves the temperature of each break point was several degrees lower than in 39 degrees C-cell lipids and nearly the same as in cells fully acclimated to low temperature (48 h or more at 15 degrees C). Fluorescence polarization studies on mixtures of natural lipids from 39 degrees C-cells and cells shifted to 15 degrees C or on mixtures of natural and synthetic lipids revealed that each of the two break points in a polarization vs. temperature plot can respond to changes in lipid composition independently of the other. It is concluded that the expeditious desaturation of certain key fatty acids, perhaps coupled with limited retailoring of phospholipid molecular species, leads to pronounced physical changes in Tetrahymena microsomal membranes as the first step of low temperature acclimation.

Acclimatization↗

Studies on thermal adaptation in Tetrahymena membrane lipids. Changes in positional distribution of fatty acids in diacyl-phospholipids and alkyl-acyl-phospholipids during temperature acclimation.

The positioning of acyl chains in both 1-O-alkyl-2-acyl- and 1,2-diacyl-phospholipids was analyzed at various time intervals for a thermotolerant strain (NT-1) of Tetrahymena pyriformis cells during cold acclimation. During the 10 h period of adaptation, cells were not able to grow but maintained the ability to divide. The content of palmitate (16 : 0) in phosphatidylcholine and phosphatidylethanolamine was decreased after temperature-shift, with a concurrent increase of palmitoleate (16 : 1 delta 9) and gamma-linolenate (18 : 3 delta 6,9,12). An increase in gamma-linolenate at the 1-position and linoleate at the 2-position was observed in diacyl-phospholipids (phosphatidylethanolamine, phosphatidylcholine and 2-aminoethylphosphonolipid). The 2-position of 1-O-alkyl-2-acyl-phosphatidylcholine and 1-O-alkyl-2-acyl-(2-aminoethyl)phosphonolipid was occupied mainly by gamma-linolenate together with cilienate (18 : 2 delta 6,11) and linoleate (18 : 2 delta 9,12). Cilienate and gamma-linolenate at the 2-position of 1-O-alkyl-2-acyl-phosphatidylcholine were increased after temperature shift, with a small decrease of linoleate. There are little significant changes in alkyl ether lipid content of phosphatidylcholine and 2-aminoethylphosphonolipid after temperature shift. The results indicate that phosphatidylethanolamine, which is most abundant and present only in the diacyl form, would play a crucial role in thermal adaptation of membrane lipids, by replacing palmitate with gamma-linolenate at its 1-position, and also that hexadecyl/gamma-linolenoyl phosphatidylcholine would be an important molecular species in the acclimation.

Animals↗

The effects of temperature acclimation on membrane sterols and phospholipids of Neurospora crassa.

Experiments were conducted to examine the effects of temperature acclimation on sterol and phospholipid biosynthesis in Neurospora crassa. Cultures grown at high (37 degrees C) and low (15 degrees C) temperatures show significant differences in free and total sterol content, sterol/phospholipid ratios and distribution of major phospholipid species in total lipids and two functionally distinct membrane fractions. The ratio of free sterols to phospholipids in total cellular lipids from 15 degrees C cultures was found to be about one-half that found at 37 degrees C, whereas sterol/phospholipid ratios of mitochondrial and microsomal membranes were found to be higher at the low growth temperature. Total sterol and phospholipid biosynthetic rates showed parallel reductions in cultures acclimating to a shift from 37 to 15 degrees C growth conditions. Distribution of [14C]acetate label into free sterols was significantly lower under these conditions, however; indicating an increase in the conversion rate of sterols to sterol esters at the lower temperature. Mitochondrial and microsomal membrane fractions showed distinct phospholipid distributions which also differed from total lipid distributions at the two growth temperatures. In each case there was a consistent decrease in phosphatidylcholine and a corresponding increase in phosphatidylethanolamine as growth temperatures were lowered.

Acclimatization↗

Changed lateral migration of phospho-LHCII in the thylakoid membrane upon acclimation of spinach to low temperatures.

Movement of proteins along the plant thylakoid membrane is of importance for several physiological events, such as state transitions and turnover and repair of the photosystem II complex. Such lateral migrations are impaired at low temperatures, which could contribute to the increased sensitivity of plants to photoinhibitory damages at low temperatures. The migration behaviour of phospho-LHCII in thylakoid membranes isolated from cold-acclimated spinach was studied and compared to that in control membranes. The rate of migration of phospho-LHCII at low temperatures is increased 2- to 3-fold and the apparent activation energy of the migration is decreased after the cold acclimation.

Acclimatization↗