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Major differences observed in transcript profiles of blueberry during cold acclimation under field and cold room conditions.

Our laboratory has been working toward increasing our understanding of the genetic control of cold hardiness in blueberry (Vaccinium section Cyanococcus) to ultimately use this information to develop more cold hardy cultivars for the industry. Here, we report using cDNA microarrays to monitor changes in gene expression at multiple times during cold acclimation under field and cold room conditions. Microarrays contained over 2,500 cDNA inserts, approximately half of which had been picked and single-pass sequenced from each of two cDNA libraries that were constructed from cold acclimated floral buds and non-acclimated floral buds of the fairly cold hardy cv. Bluecrop (Vaccinium corymbosum L.). Two biological samples were examined at each time point. Microarray data were analyzed statistically using t tests, ANOVA, clustering algorithms, and online analytical processing (OLAP). Interestingly, more transcripts were found to be upregulated under cold room conditions than under field conditions. Many of the genes induced only under cold room conditions could be divided into three major types: (1) genes associated with stress tolerance; (2) those that encode glycolytic and TCA cycle enzymes, and (3) those associated with protein synthesis machinery. A few of the genes induced only under field conditions appear to be related to light stress. Possible explanations for these differences are discussed in physiological context. Although many similarities exist in how plants respond during cold acclimation in the cold room and in the field environment, there are major differences suggesting caution should be taken in interpreting results based only on artificial, cold room conditions.

Acclimatization↗

Acclimation of Arabidopsis thaliana to the light environment: the role of photoreceptors.

The regulation by light of the composition of the photosynthetic apparatus was investigated in photomorphogenic mutants of Arabidopsis thaliana (L.) Heynh. cv. Landsberg erecta. Leaf chlorophyll, photosynthesis, photosystem II function, and ribulose-1, 5-bisphosphate carboxylase-oxygenase and photosystem II contents were determined for plants grown under high- or low-irradiance growth regimes. Although certain mutant lines had altered chloroplast composition compared to the wild type, all photoreceptor mutants tested were capable of light-dependent changes in chloroplast composition and photosynthetic function, indicating that photoreceptors do not play a central role in the regulation of acclimation at the level of the chloroplast. However, the clear acclimation defect in a det1 signal transduction mutant indicates that photoreceptor-controlled responses either share regulatory components with acclimation, or are important in the expression of components which in turn regulate acclimation. We suggest that the COP/DET/FUS regulatory cluster is a focus for multiple signal transduction pathways, including some of the metabolic signals which form the basis for the acclimatory response.

Arabidopsis↗

Lichens show that fungi can acclimate their respiration to seasonal changes in temperature.

Five species of lichens, the majority members of a soil-crust community ( Cladonia convoluta, Diploschistes muscorum, Fulgensia fulgens, Lecanora muralis, Squamarina lentigera) showed seasonal changes of temperature sensitivity of their dark respiration (DR) to such an extent that several substantially met the definition of full acclimation, i.e. near identical DR under different nocturnal temperature conditions during the course of the year. C. convoluta, for example, had maximal DR at 5 degrees C of -0.42, -1.11 and -0.09 nmol CO(2) g(-1) s(-1) in autumn, winter, and summer, respectively, a tenfold range. However, at the mean night temperatures for the same three seasons, 9.7 degrees C, 4.2 degrees C and 13.6 degrees C, maximal DR were almost identical at -1.11, -0.93, and -1.45 nmol CO(2) g(-1) s(-1). The information was extracted from measurements using automatic cuvettes that continuously recorded a sample lichen's gas exchange every 30 min under near-natural conditions. The longest period (for L. muralis) covered 15 months and 22,000 data sets whilst, for the other species studied, data blocks were available throughout the calendar year. The acclimation of DR means that maximal net carbon fixation rates remain substantially similar throughout the year and are not depressed by increased carbon loss by respiration in warmer seasons. This is especially important for lichens because of their normally high rate of DR compared to net photosynthesis. We suggest that lichens, especially soil-crust species, could be a suitable model for fungi generally, a group of organisms for which little is known about temperature acclimation because of the great difficulty in separating the organism from its growth medium. Fungi, whether saprophytic, symbiotic or parasitic, including soil lichens, are important components of soil ecosystems and contribute much of the respired CO(2) from these systems. Temperature acclimation by fungi would mean that expected increases in carbon losses caused by global climate warming from soil ecosystems might not be as extensive as first thought. This would ameliorate this positive feedback loop present in some climate models and might substantially lower the predicted warming.

Acclimatization↗

Tissue distribution, effects of salinity acclimation, and ontogeny of aquaporin 3 in the marine teleost, silver sea bream (Sparus sarba).

The purpose of the present study was to ascertain the tissue-specific expression of the water channel protein, aquaporin 3 (AQP3), during salinity acclimation and larval development of silver sea bream (Sparus sarba). A cDNA fragment encoding aquaporin 3 (aqp3) from silver sea bream gill was cloned and from the deduced amino acid sequence a polyclonal antibody was prepared. AQP3 was found to be present in gill, kidney, liver, brain, heart, and spleen but not in whole blood. The abundance of AQP3 was significantly highest in gills of hypoosmotic (6 ppt) and isoosmotic (12 ppt) acclimated sea bream when compared to seawater (33 ppt) and hypersaline (50 ppt)- acclimated sea bream. Spleen tissue also displayed significantly high levels of AQP3 protein in hypoosmotic and isoosmotic salinities whereas the AQP3 abundance in brain, liver, heart, and kidney remained unchanged across the range of salinities tested. The ontogenetic profile of AQP3 was also investigated from developing sea bream larvae and AQP3 was first detected at 14 days posthatch (dph) and increased steadily up to 28-46 dph. In conclusion, this study has demonstrated that AQP3 expression is modulated in gill and spleen tissue of salinity acclimated sea bream and that it can be detected relatively early during larval development.

Acclimatization↗

Gene expression profiles during heat acclimation in Arabidopsis thaliana suspension-culture cells.

Thermotolerance is induced by moderated heat acclimation. Suspension cultures of heat-acclimated Arabidopsis thaliana L. (Heynh.), ecotype Columbia, show thermotolerance against lethal heat shock (9 min, 50 degrees C), as evidenced by a chlorophyll assay and fluorescein diacetate staining. To monitor the genome-wide transcriptome changes induced by heat acclimation at 37 degrees C, we constructed an A. thaliana cDNA microarray containing 7,989 unique genes, and applied it to A. thaliana suspension-culture cells harvested at various times (0.5, 1, 2.5, 6, and 16 h) during heat acclimation. Data analysis revealed 165 differentially expressed genes that were grouped into ten clusters. We compared these genes with published and publicly available microarray heat-stress-related data sets in AtGenExpress. Heat-shock proteins were strongly expressed, as previously reported, and we found several of the up-regulated genes encoded detoxification and regulatory proteins. Moreover, the transcriptional induction of DREB2 (dehydration responsive element-binding factor 2) subfamily genes and COR47/rd17 under heat stress suggested cross-talk between the signaling pathways for heat and dehydration responses.

Acclimatization↗

Biodegradation kinetics of 2,4,6-trichlorophenol by an acclimated mixed microbial culture under aerobic conditions.

The objective of this study was to achieve a better quantitative understanding of the kinetics of 2,4,6-trichlorophenol (TCP) biodegradation by an acclimated mixed microbial culture. An aerobic mixed microbial culture, obtained from the aeration basin of the wastewater treatment plant, was acclimated in shake flasks utilizing various combinations of 2,4,6-TCP (25-100 mg l(-1)), phenol (300 mg l(-1)) and glycerol (2.5 mg l(-1)) as substrates. Complete primary TCP degradation and a corresponding stoichiometric release of chloride ion were observed by HPLC and IEC analytical techniques, respectively. The acclimated cultures were then used as an inoculum for bench scale experiments in a 4 l stirred-tank reactor (STR) with 2,4,6-TCP as the sole carbon/energy (C/E) source. The phenol acclimated mixed microbial culture consisted of primarily Gram positive and negative rods and was capable of degrading 2,4,6-TCP completely. None of the predicted intermediate compounds were detected by gas chromatography in the cell cytoplasm or supernatant. Based on the disappearance of 2,4,6-TCP, degradation was well modelled by zero-order kinetics which was also consistent with the observed oxygen consumption. Biodegradation rates were compared for four operating conditions including two different initial 2,4,6-TCP concentrations and two different initial biomass concentrations. While the specific rate constant was not dependent on the initial 2,4,6-TCP concentration, it did depend on the initial biomass concentration (X (init)). A lower biomass concentration gave a much higher zero-order specific degradation rate. This behaviour was attributed to a lower average biomass age or cell retention time (theta(x)) for these cultures. The implications of this investigation are important for determining and predicting the potential risks associated with TCP, its degradation in the natural environment or the engineering implications for ex situ treatment of contaminated ground water or soil.

Acclimatization↗

Effects of chlorophyllide a oxygenase overexpression on light acclimation in Arabidopsis thaliana.

Land plants change the compositions of light-harvesting complexes (LHC) and chlorophyll (Chl) a/b ratios in response to the variable light environments which they encounter. In this study, we attempted to determine the mechanism which regulates Chl a/b ratios and whether the changes in Chl a/b ratios are essential in regulation of LHC accumulation during light acclimation. We hypothesized that changes in the mRNA levels for chlorophyll a oxygenase (CAO) involved in Chl b biosynthesis are an essential part of light response of Chl a/b ratios and LHC accumulation. We also examined the light-intensity dependent response of CAO-overexpression and wild-type Arabidopsis thaliana plants. When wild-type plants were acclimated from low-light (LL) to high-light (HL) conditions, CAO mRNA levels decreased and the Chl a/b ratio increased. In transgenic plants overexpressing CAO, the Chl a/b ratio remained low under HL conditions; thereby suggesting that changes in the CAO mRNA levels are necessary for those in Chl a/b ratios upon light acclimation. Under HL conditions, the accumulation of Lhcb1, Lhcb3 and Lhcb6 was enhanced in plants overexpressing CAO. On the contrary, in a CAO-deficient mutant, chlorina 1-1, theaccumulation of Lhcb1, Lhcb2, Lhcb3, Lhcb6 and Lhca4 was reduced. In comparison to wild-type, beta-carotene levels were reduced in CAO-overexpressing plants, while they were elevated in chlorina 1-1 mutants. These results imply that the transcriptional control of CAO is a part of the regulatory mechanism for the accumulation of a distinct set of LHC proteins upon light acclimation.

Acclimatization↗

Influences of subzero thermal acclimation on mitochondrial membrane composition of temperate zone marine bivalve mollusks.

The phospholipid and phospholipid fatty acid composition of gill mitochondrial membranes from two temperate zone marine bivalve mollusks, the quahog, Mercenaria mercenaria, and the American oyster, Crassostrea virginica, were examined after acclimation to 12 and -1 degree C. Cardiolipin (CL) was the only phospholipid with proportions altered upon acclimation to -1 degree C, increasing 188% in the mitochondrial membranes of M. mercenaria. Although the ratio of bilayer stabilizing to destabilizing lipids is frequently associated with cold acclimation in ectothermic species, no change was found in this ratio in either of the species. Polyunsaturated fatty acids (PUFA) were found only to increase in C. virginica with cold acclimation, with total n-3 PUFA increasing in the phospholipid phosphatidylethanolamine, total n-6 PUFA increasing in CL, and total PUFA increasing in phosphatidylinositol. Monounsaturated fatty acids, not PUFA, were found to have increased in M. mercenaria, with 18:1 n-9 increasing by 150% in CL, and 20:1 increasing in both CL and phosphatidylcholine, by 146 and 192%, respectively. These manipulations of membrane phospholipid and fatty acid composition may represent an attempt by these species to help maintain membrane function at low temperatures.

Adaptation, Physiological↗

The effect of acclimation temperature on the activation energies of state III respiration and on the unsaturation of membrane lipids of goldfish mitochondria.

The influence of the acclimation temperature on the thermotropic behaviour of mitochondrial respiration and on the degree of unsaturation of mitochondrial membrane lipids has been studied. The mitochondria were isolated from red muscle, white muscle and liver of goldfish acclimated to 5, 20 and 30 degrees C. ADP-activated succinate oxidation was measured at different temperatures and resulted in non-linear Arrhenius-plots with breaks between 10 and 23 degrees C. As for the break-temperatures, there was found a shift downwards in preparations of decreased acclimation temperatures. This could be caused by a changed composition of membrane lipids and a simultaneous shift of the membrane phase transition temperature. Therefore, the fatty acid composition of all membrane preparations was analyzed. However, no consistent change of the degree of unsaturation due to a changed acclimation temperature could be found.

Acclimatization↗

Cold acclimation and resistance to ethanol-induced hypothermia.

The fall in core temperature induced by a fixed dose of ethanol (1 g x kg-1 i.p.) was an exponential function of the ambient temperature over the range 0-18 degrees C. In rats acclimated to 4 degrees C for 7 days the dose response curve relating the fall in body temperature to ethanol was markedly attenuated. The hypothermic effect of ethanol declined exponentially over 20 days of exposure to 4 degrees C and by the 20th day the fall was no greater than in saline injected controls. Blood ethanol concentrations were similar in acclimated and non-acclimated rats indicating that pharmacokinetic factors do not account for the altered responses. Although the time course of the development of resistance to the hypothermic effect of ethanol parallels that of cold acclimation to 4 degrees C, and the development of non-shivering thermogenesis, the attenuated drug effect does not appear to be due to the altered metabolic activity of brown adipose tissue. It is suggested that the modulation of the effect of ethanol in lowering the thermoregulatory set point results from central nervous system adaptation to the environmental thermal stress.

Acclimatization↗

Cold acclimation of obese (ob/ob) mice: effects of energy balance.

Obese (ob/ob) and lean mice at 4 weeks of age were housed at 23 degrees C or 14 degrees C for 4 to 8 weeks to examine effects of acclimation to mild cold on energy balance. Energy intake of young lean mice increased by about 50% when housed at 14 degrees C, but energy intake of cold-acclimated obese mice increased by only 8%. Efficiency of energy retention (ratio of energy gain to energy intake) in obese mice declined from 22% +/- 1.2% at 23 degrees C to 10% +/- 1.8% after 4 weeks at 14 degrees C. Lean mice exhibited a less pronounced response to temperature; their efficiency of energy retention declined from 7% +/- 1.3% at 23 degrees C to 4% +/- 2.2% after 4 weeks at 14 degrees C. After 8 weeks of cold exposure, body weights and efficiency of energy retention became equal in obese and lean mice. Calculated heat production of cold-acclimated obese and lean mice was 40% higher than that of respective controls. Obese mice reacclimated to 23 degrees C after being kept for 4 weeks at 14 degrees C consumed the same amount of energy and were 16% more efficient than obese maintained at 23 degrees C; reacclimated lean mice consumed 12% more energy but were 53% less efficient than lean mice maintained at 23 degrees C. The results indicate that obese mice are able to increase heat production and markedly reduce their efficiency of energy retention when acclimated to mild cold but that they, unlike lean mice, rapidly revert to a high efficiency of energy retention after 4 weeks of reacclimation to 23 degrees C.

Acclimatization↗

Regulation of submaxillary gland muscarinic receptors during heat acclimation.

Binding properties of submaxillary gland muscarinic receptors and agonist-induced saliva secretion were studied in rats subjected to heat acclimation. The maximal binding capacity for the muscarinic antagonist N-[3H]methyl-4-piperidyl benzilate was increased from control value of 0.21 to 0.40 pmol/mg protein within 1-2 days of heat acclimation. The increase in the number of muscarinic receptors per gland (100%) was by far higher than the increase in tissue weight (20%), indicating higher density of receptors in the acinar cells of the treated rats. High levels of receptors coincided with the appearance of high-affinity binding sites for muscarinic agonists (oxotremorine, pilocarpine and carbamylcholine), and with reduced tissue sensitivity to pilocarpine. After 4-8 weeks of heat acclimation, the number of receptors as well as tissue response to pilocarpine returned to control levels. These results suggest a functional correlation between the transient upregulation muscarinic receptors in the submaxillary gland and the physiological activity in salivary secretion, and indicate that the high-affinity muscarinic receptors may attenuate saliva secretion during the initial phase of heat acclimation.

Acclimatization↗

Plasma opioid peptide responses during heat acclimation in humans.

Plasma beta-endorphin, Met-enkephalin and Peptide F immunoreactivity (ir) were measured at rest and following exercise on three days (days 1, 4, 8) of an eight day heat acclimation regime. Fourteen male subjects demonstrated physiological heat acclimation adaptations. Our data demonstrated a differential response of peripheral plasma levels of endogenous opioid peptides (EOP) to exercise in the heat. In addition, EOP did not follow the same time-course of other physiological adaptations as no differences (day 1 vs. 4 vs. 8) in resting or exercise levels were observed over the eight day heat acclimation regime. Significant increases in beta-endorphin ir (pre- to post-exercise) appear to reflect concomitant exercise-heat related changes. The increased peripheral levels of beta-endorphin were correlated to plasma levels of cortisol. Heat and exercise stress may result in a reduction of Met-enkephalin ir observed in peripheral plasma and might be due to degradation or a decrease in processing from the larger precursors. The differential responses of EOP suggest the possibility of separate physiological roles for these peptides during exercise in the heat but peripheral plasma levels of EOP do not appear to reflect acute heat acclimation changes.

Acclimatization↗

The participation of shivering and nonshivering thermogenesis in warm and cold-acclimated rats.

1. Both shivering and nonshivering thermogenesis are ways of increasing oxygen consumption in the rat. The participation of nonshivering thermogenesis after cold acclimation is well established, but there is little evidence of the relative importance of each kind of thermogenesis in acclimated and nonacclimated rats. 2. Oxygen consumption and rectal temperature were measured in warm and cold-acclimated rats exposed both to cold (10 degrees C) and warm (30 degree C) environments. 3. Shivering was blocked by mephenesin (10 mg/100 g body wt) and nonshivering thermogenesis was blocked by propranolol (0.5 mg/100 g body wt). 4. Control animals were injected with saline (0.5 ml/100 g body wt). 5. The results showed that shivering is not effective in maintaining a constant rectal temperature and that nonshivering thermogenesis does not contribute to the increase in oxygen consumption of warm-acclimated rats exposed to cold.

Acclimatization↗

The effect of photoperiod on changes in plasma, glucose, cholesterol, and free fatty acids during cold acclimation in frogs.

1. Many previous experiments on cold acclimation in frogs were carried out on animals maintained in total darkness. I tested the effect of photoperiod on changes in plasma FFA, glucose, and cholesterol during cold acclimation. 2. Hematocrit levels were lower whereas plasma FFA levels were higher in frogs kept in total darkness during cold acclimation. 3. Plasma glucose and plasma cholesterol levels were not affected by photoperiod during cold acclimation.

Acclimatization↗

Thermoregulatory activity in the rat: effects of hypohydration, hypovolemia and hypertonicity and their interaction with short-term heat acclimation.

Hypothalamic temperature thresholds to heat-induced (40 degrees C ambient temperature) tail vasodilation (Vth) and salivation (Sth) as well as salivary flow rate and volume were studied in conscious rats, hypohydrated (24 hr water deprivation), hypovolemic (20% dextran sc), hypertonic (1M NaCL po), hypertonic and hypovolemic and heat-acclimated (5 days at 34 degrees C) before and after hypohydration. Sth was elevated in hypohydrated, hypovolemic, hypertonic and heat-acclimated hypohydrated rats concomitantly with a remarkable decrease in saliva volume, flow rate and heat tolerance. Heat acclimation alone resulted in a reduction in Vth, Sth, salivary flow and volume. Vth was not affected by hypohydration, but was elevated following hypovolemia and combined hypovolemia and hypertonicity. It is concluded that alterations in both plasma volume and osmolarity, which may occur during hypohydration, play a major role in the alteration in thermoregulatory responses during hypohydration. Heat acclimation does not improve tolerance during hypohydration. Thus, during hypohydration, the control of body fluids overrides thermoregulation.

Acclimatization↗

Comparison of thermal acclimation effects on the metabolism of Chalcides ocellatus (desert lizard) and Lacerta vivipara (cool-temperate lizard).

The ocellated lizard, Chalcides ocellatus, and the common lizard, Lacerta vivipara, were acclimated to two temperature regimes (10 and 30 degrees C with 12L:12D photoperiod) for 2 weeks. The oxygen consumption rates were measured for both species at temperatures between 5 and 35 degrees C. Cold acclimation was observed in L. vivipara and was in the form of an upward translation of the metabolic rate-temperature curve in comparison to the warm-acclimated lizards (30 degrees C). Chalcides ocellatus showed a response to cold acclimation by decreasing its metabolism only at 10 degrees C.

Acclimatization↗

Fluid compartmentation in skeletal muscle and carcass of Mus musculus acclimated to water scarcity.

1. Total water (TW), and extracellular water (ECW) (as sodium and chloride space) were determined in skeletal muscle and carcass of Mus musculus acclimated to long-term water shortage. 2. The presence of fat in control mice and those in early stages of acclimation resulted in an apparent increase in TW and ECW as acclimation proceeded. 3. In contrast, fluid volumes per fat-free weight were either unchanged from controls or reduced. 4. Sodium space exceeded chloride space. 5. Muscle and carcass had essentially the same pattern of fluid shifts. 6. We conclude that ECW maintenance is a preeminent component of the acclimation process in this species.

Adaptation, Physiological↗