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Inhibition and acclimation of photosynthesis to heat stress is closely correlated with activation of ribulose-1,5-bisphosphate Carboxylase/Oxygenase

Increasing the leaf temperature of intact cotton (Gossypium hirsutum L.) and wheat (Triticum aestivum L.) plants caused a progressive decline in the light-saturated CO2-exchange rate (CER). CER was more sensitive to increased leaf temperature in wheat than in cotton, and both species demonstrated photosynthetic acclimation when leaf temperature was increased gradually. Inhibition of CER was not a consequence of stomatal closure, as indicated by a positive relationship between leaf temperature and transpiration. The activation state of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco), which is regulated by Rubisco activase, was closely correlated with temperature-induced changes in CER. Nonphotochemical chlorophyll fluorescence quenching increased with leaf temperature in a manner consistent with inhibited CER and Rubisco activation. Both nonphotochemical fluorescence quenching and Rubisco activation were more sensitive to heat stress than the maximum quantum yield of photochemistry of photosystem II. Heat stress led to decreased 3-phosphoglyceric acid content and increased ribulose-1, 5-bisphosphate content, which is indicative of inhibited metabolite flow through Rubisco. We conclude that heat stress inhibited CER primarily by decreasing the activation state of Rubisco via inhibition of Rubisco activase. Although Rubisco activation was more closely correlated with CER than the maximum quantum yield of photochemistry of photosystem II, both processes could be acclimated to heat stress by gradually increasing the leaf temperature.

Journal Article↗

Chloroplast acclimation in leaves of Guzmania monostachia in response to high light.

Acclimation of leaves to high light (HL; 650 micromol m(-2) s(-1)) was investigated in the long-lived epiphytic bromeliad Guzmania monostachia and compared with plants maintained under low light (LL; 50 micromol m(-2) s(-1)). Despite a 60% decrease in total chlorophyll in HL-grown plants, the chlorophyll a/b ratio remained stable. Additionally, chloroplasts from HL-grown plants had a much lower thylakoid content and reduced granal stacking. Immunofluorescent labeling techniques were used to quantify the level of photosynthetic polypeptides. HL-grown plants had 30% to 40% of the content observed in LL-grown plants for the light-harvesting complex associated with photosystems I and II, the 33-kD photosystem II polypeptide, and Rubisco. These results were verified using conventional biochemical techniques, which revealed a comparable 60% decrease in Rubisco and total soluble protein. When expressed on a chlorophyll basis, the amount of protein and Rubisco was constant for HL- and LL-grown plants. Acclimation to HL involves a tightly coordinated adjustment of photosynthesis, indicating a highly regulated decrease in the number of photosynthetic units manifested at the level of the content of light-harvesting and electron transport components, the amount of Rubisco, and the induction of Crassulacean acid metabolism. This response occurs in mature leaves and may represent a strategy that is optimal for the resource-limited epiphytic niche.

Chlorophyll↗

Acclimation of Photosynthetic and Respiratory Carbon Dioxide Exchange to Growth Temperature in Atriplex lentiformis (Torr.) Wats.

Atriplex lentiformis plants collected from coastal and desert habitats exhibit marked differences in capacity to adjust photosynthetic response to changes in growth temperature. Plants from desert habitats grown at 43 C day/30 C night temperatures had higher CO(2) uptake rates at high temperatures but reduced rates at low temperatures as compared to plants grown at 23 C day/18 C night temperatures. In contrast, growth of the coastal plants at high temperatures resulted in markedly reduced photosynthetic rates at all measurement temperatures.Leaf conductances to CO(2) were not important in controlling either the differences in the temperature dependence of net CO(2) uptake or the differences in photosynthetic capacities at any measurement temperature. At low measurement temperatures, differences in photosynthetic capacities among plants acclimated to the contrasting growth regimes were correlated with differences in leaf ribulose diphophate carboxylase activities. At high measurement temperatures, the improved net photosynthetic performance of the high temperature acclimated desert plants appeared to be due to a combination of decreased respiration rates, decreased temperature dependence of respiration, and an apparent increased thermal stability of photosynthetic CO(2) exchange.

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Photosynthetic Acclimation to Temperature in the Desert Shrub, Larrea divaricata: I. Carbon Dioxide Exchange Characteristics of Intact Leaves.

Larrea divaricata, a desert evergreen shrub, has a remarkable ability to adjust its photosynthetic temperature response characteristics to changing temperature conditions. In its native habitat on the floor of Death Valley, California, plants of this C(3) species when provided with adequate water are able to maintain a relatively high and constant photosynthetic activity throughout the year even though the mean daily maximum temperature varies by nearly 30 C from winter to summer. The temperature dependence of light-saturated net photosynthesis varies in concert with these seasonal temperature changes whereas the photosynthetic rate at the respective optimum temperatures shows little change.Experiments on plants of the same age, grown at day/night temperatures of 20/15, 35/25, and 45/33 C with the same conditions of day length and other environmental factors, showed a similar photosynthetic acclimation response as observed in nature. An analysis was made of a number of factors that potentially can contribute to the observed changes in the temperature dependence of net CO(2) uptake at normal CO(2) and O(2) levels. These included stomatal conductance, respiration, O(2) inhibition of photosynthesis, and nonstomatal limitations of CO(2) diffusive transport. None of these factors, separately or taken together, can account for the observed acclimation responses. Measurements under high saturating CO(2) concentrations provide additional evidence that the observed adaptive responses are primarily the result of changes in intrinsic characteristics of the photosynthetic machinery at the cellular or subcellular levels. Two apparently separate effects of the growth temperature regime can be distinguished: one involves an increased capacity for photosynthesis at low, rate-limiting temperatures with decreased growth temperature, and the other an increased thermal stability of key components of the photosynthetic apparatus with increased growth temperature.

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Cold Acclimation of Suspension Cultures of Pinus sylvestris in Response to Light and Temperature Treatments.

Suspension cultures of Pinus sylvestris L. (Provenance Södra Ydre) were used to determine frost hardiness after manipulating daylengths and temperatures. Frost hardiness was determined with the triphenyltetrazolium chloride (TTC) reduction method. The cultures were able to acclimate and increase frost hardiness levels; both low temperature (2 degrees C) and short day (8 h) treatments were used at the same time, but increased survival temperatures were not achieved when only one type of stimulus was used. Inasmuch as intact seedlings can be partially acclimated by a single type of stimulus, the results indicate that the organization of the cells to tissues plays a role for the hardening process in vivo.

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Microsomal Phospholipid Molecular Species Alterations during Low Temperature Acclimation in Dunaliella.

A detailed analysis of the low temperature-induced alterations of Dunaliella salina (UTEX 1644) microsomal membrane lipids was carried out. Microsomal membranes were isolated from cells grown at 30 degrees C, from cells shifted to 12 degrees C for 12 hours, and from cells acclimated to 12 degrees C. Fatty acid analyses of the major lipid classes demonstrated significant changes in the fatty acid composition of phosphatidylcholinemine (PE) and phosphatidylglycerol (PG) but not phosphatidylcholine (PC) during the initial 12 hours at low temperature. These changes did not entail enhanced desaturation of linoleic acid. Subsequent to 12 hours, the proportions of linolenic acid increased in all phospholipids.Molecular species analyses of the phospholipids demonstrated that the most immediate changes following a shift to low temperature were limited to several molecular species of PE and PG. The changes observed in PE included a decrease in C(30) species and concomitant increases in C(34) and C(36) species. Compositional changes associated with PG entailed the emergence of a new molecular species (18:1/18:1) not found at 30 degrees C. The retailoring of molecular species resulted in an increase in the number of species having two unsaturated acyl chains and did not reflect a simple enhancement of desaturase activity as suggested by the fatty acid analysis. We conclude that the initial alterations in response to low temperature stress involve discrete changes in certain molecular species. These and further alterations of molecular species following acclimation to low temperature would appear to augment increases in acyl chain desaturation as a means of modifying membrane properties in response to low temperature stress.

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Protein and Lipid Compositions of Isolated Plasma Membranes from Orchard Grass (Dactylis glomerata L.) and Changes during Cold Acclimation.

The chemical composition of plasma membrane fractions isolated from orchard grass seedlings (Dactylis glomerata L.) was analyzed and compared with endomembranes. The plasma membrane is characterized by an enrichment of sterols and a lower degree of unsaturation of phospholipids. Steryl glycosides were found to be one of the lipid components of the plasma membrane, but steryl esters and galactolipids were barely detectable. Diphosphatidyl glycerol was characteristically detected in the mitochondrial membrane, but not in the plasma membrane fraction. Plasma mambrane fraction was also characterized by its ;lower fluidity' in comparison with the endomembranes. This may be due to the large amount of sterols and the lower degree of phospholipid unsaturation in plasma membranes.Electrophoretic comparison of polypeptides was also made between different membranes. The distribution patterns of polypeptides revealed on one- and two-dimensional SDS-slab gels were characteristic for those membranes. The presence of glycopeptide complements was a useful criterion for distinguishing plasma membrane from other membranes. The plasma membrane and the ER + Golgi membranes were enriched in glycopeptides. However, a marked difference was revealed in the total number and the molecular weights of the peptides.During cold acclimation of orchard grass seedlings, the degree of fatty acid unsaturation increased only slightly in plasma membrane, unlike in endomembranes. The change in sterols-to-phospholipids ratio in plasma membrane was also slight. On the other hand, the phospholipid-to-protein ratio increased significantly in the plasma membrane as cold hardiness increased. A significant change in the polypeptide complements of plasma membrane was also demonstrated during cold acclimation.

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Chemical and Biophysical Changes in the Plasma Membrane during Cold Acclimation of Mulberry Bark Cells (Morus bombycis Koidz. cv Goroji).

The lipid and protein composition of the plasma membrane isolated from mulberry (Morus bombycis Koidz.) bark cells was analyzed throughout the cold acclimation period under natural and controlled environment conditions. There was a significant increase in phospholipids and unsaturation of their fatty acids during cold acclimation. The ratio of sterols to phospholipids decreased with hardiness, primarily due to the large increase in phospholipids. The fluidity of the plasma membrane, as determined by fluorescent polarization technique, increased with hardiness. Electrophoresis of plasma membrane proteins including glycoproteins revealed change in banding pattern during the early fall to winter period. Some of the protein changes could be related to growth cessation and defoliation. However, minor changes in proteins also occurred during the most active period of hardening. Changes in glycoproteins were coincident both with changes in growth stages and with the development of cold hardiness.

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Metabolic Acclimation to Anoxia Induced by Low (2-4 kPa Partial Pressure) Oxygen Pretreatment (Hypoxia) in Root Tips of Zea mays.

Young intact plants of maize (Zea mays L. cv INRA 508) were exposed to 2 to 4 kilopascals partial pressure oxygen (hypoxic pretreatment) for 18 hours before excision of the 5 millimeter root apex and treatment with strictly anaerobic conditions (anoxia). Hypoxic acclimation gave rise to larger amounts of ATP, to larger ATP/ADP and adenylate energy charge ratios, and to higher rates of ethanol production when excised root tips were subsequently made anaerobic, compared with root tips transferred directly from aerobic to anaerobic media. Improved energy metabolism following hypoxic pretreatment was associated with increased activity of alcohol dehydrogenase (ADH), and induction of ADH-2 isozymes. Roots of Adh1(-) mutant plants lacked constitutive ADH and only slowly produced ethanol when made anaerobic. Those that were hypoxically pretreated acclimated to anoxia with induction of ADH2 and a higher energy metabolism, and a rate of ethanol production comparable to that of nonmutants. All these responses were insensitive to the presence or absence of NO(3) (-). Additionally, the rate of ethanol production was about 50 times greater than the rate of reduction of NO(3) (-) to NO(2) (-). These results indicate that nitrate reductase does not compete effectively with ADH for NADH, or contribute to energy metabolism during anaerobic respiration in this tissue through nitrate reduction. Unacclimated root tips of wild type and Adhl(-) mutants appeared not to survive more than 8 to 9 hours in strict anoxia; when hypoxically pretreated they tolerated periods under anoxia in excess of 22 hours.

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Molecular cloning and relationship to freezing tolerance of cold-acclimation-specific genes of alfalfa.

Cold-acclimation-specific (CAS) gene expression has been examined by screening a cDNA library prepared from poly(A)(+) RNA of cold-acclimated seedlings of a freezing-tolerant variety of alfalfa (Medicago falcata cv Anik). Three CAS cDNA clones, pSM784, pSM2201, and pSM2358, representing different sequence species, have been used to investigate the relative abundance and time-course of accumulation of corresponding transcripts. Results obtained show that the expression of these CAS genes is regulated in a coordinated manner most likely at the level of transcription. The expression of genes, as measured by mRNA abundance corresponding to the three CAS cDNA clones, is not stimulated or induced by heat shock, water stress, abscisic acid, or wounding. A positive correlation is observed between the expression of these cloned sequences and the degree of freezing-tolerance in four alfalfa cultivars.

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Tubulin Isotypes in Rye Roots Are Altered during Cold Acclimation.

The cold stability of cortical microtubules in root-tip cells of winter rye (Secale cereale L. cv Puma) is altered by growth temperature (GP Kerr, JV Carter [1990] Plant Physiol 93:77-82). One hypothesis for the basis of this alteration is that different tubulin isotypes are present at different growth temperatures, and that the cold stability of microtubules is affected by these isotypic differences. We have explored the first part of this hypothesis by comparing protein extracts from roots of seedlings grown for 2 days at 22 degrees C (nonacclimated) or for an additional 2 or 4 days at 4 degrees C (cold-acclimated). Immunoblots of two-dimensional polyacrylamide gels were probed with monoclonal antibodies to alpha- and beta-tubulin. At least six alpha- and seven beta-tubulins were present in the extracts from both the nonacclimated and cold-acclimated roots. Changes in electrophoretic mobility and isotype number of both alpha- and beta-tubulin were observed after only 2 days at 4 degrees C. Further changes in tubulin were observed after 4 days at 4 degrees C. Changes in alpha-tubulin were more pronounced than those in beta-tubulin.

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Cold acclimation in Arabidopsis and wheat : a response associated with expression of related genes encoding ;boiling-stable' polypeptides.

Changes in gene expression occur during cold acclimation in a wide variety of plant species. Here we show that a number of the polypeptides encoded by cold-regulated (cor) genes of Arabidopsis thaliana L. (Heyn) and wheat share the unusual biochemical property that they remain soluble upon boiling in aqueous solution. Further, cDNA cloning in conjunction with Southern and Northern analyses indicate that wheat has a cor gene that is related to Arabidopsis cor47, a gene encoding a 47 kilodalton ;boiling-stable' COR polypeptide. We suggest it is likely that the boiling-stable COR polypeptides have a fundamental role in plants acclimating to cold temperatures and discuss the possibility that they may act as cryoprotectants.

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Mutations in the Ca2+/H+ transporter CAX1 increase CBF/DREB1 expression and the cold-acclimation response in Arabidopsis.

Transient increases in cytosolic free calcium concentration ([Ca2+]cyt) are essential for plant responses to a variety of environmental stimuli, including low temperature. Subsequent reestablishment of [Ca2+]cyt to resting levels by Ca2+ pumps and antiporters is required for the correct transduction of the signal [corrected]. C-repeat binding factor/dehydration responsive element binding factor 1 (Ca2+/H+) antiporters is required for the correct transduction of the signal. We have isolated a cDNA from Arabidopsis that corresponds to a new cold-inducible gene, rare cold inducible4 (RCI4), which was identical to calcium exchanger 1 (CAX1), a gene that encodes a vacuolar Ca2+/H+ antiporter involved in the regulation of intracellular Ca2+ levels. The expression of CAX1 was induced in response to low temperature through an abscisic acid-independent pathway. To determine the function of CAX1 in Arabidopsis stress tolerance, we identified two T-DNA insertion mutants, cax1-3 and cax1-4, that display reduced tonoplast Ca2+/H+ antiport activity. The mutants showed no significant differences with respect to the wild type when analyzed for dehydration, high-salt, chilling, or constitutive freezing tolerance. However, they exhibited increased freezing tolerance after cold acclimation, demonstrating that CAX1 plays an important role in this adaptive response. This phenotype correlates with the enhanced expression of CBF/DREB1 genes and their corresponding targets in response to low temperature. Our results indicate that CAX1 ensures the accurate development of the cold-acclimation response in Arabidopsis by controlling the induction of CBF/DREB1 and downstream genes.

Abscisic Acid↗

Cold calcium signaling in Arabidopsis involves two cellular pools and a change in calcium signature after acclimation.

Cold shock elicits an immediate rise in cytosolic free calcium concentration ([Ca2+]cyt) in both chilling-resistant Arabidopsis and chilling-sensitive tobacco (Nicotiana plumbaginifolia). In Arabidopsis, lanthanum or EGTA caused a partial inhibition of both cold shock [Ca2+]cyt elevation and cold-dependent kin1 gene expression. This suggested that calcium influx plays a major role in the cold shock [Ca2+]cyt response and that an intracellular calcium source also might be involved. To investigate whether the vacuole (the major intracellular calcium store in plants) is involved, we targeted the calcium-dependent photoprotein aequorin to the cytosolic face of the vacuolar membrane. Cold shock calcium kinetics in this microdomain were consistent with a cold-induced vacuolar release of calcium. Treatment with neomycin or lithium, which interferes with phosphoinositide cycling, resulted in cold shock [Ca2+]cyt kinetics consistent with the involvement of inositol trisphosphate and inositide phosphate signaling in this response. We also investigated the effects of repeated and prolonged low temperature on cold shock [Ca2+]cyt. Differences were observed between the responses of Arabidopsis and N. plum-baginifolia to repeated cold stimulation. Acclimation of Arabidopsis by pretreatment with cold or hydrogen peroxide caused a modified calcium signature to subsequent cold shock. This suggests that acclimation involves modification of plant calcium signaling to provide a "cold memory."

Acclimatization↗

Photoprotection mutants of Arabidopsis thaliana acclimate to high light by increasing photosynthesis and specific antioxidants.

Biochemical and physiological acclimation to different light environments is crucial for plant growth and survival. In high light (HL), feedback de-excitation (qE) is a well-known photoprotective mechanism that dissipates excess excitation energy in the light-harvesting antenna of photosystem II (PSII) and relieves excitation pressure in the photosynthetic electron transport chain. The xanthophylls zeaxanthin (Z) and lutein (L) function in qE, but also have roles as antioxidants. Although several studies have shown that qE is important during short-term fluctuations in light intensity, here we show that it is not required for the growth of Arabidopsis thaliana in prolonged HL conditions in the laboratory. Mutants that are deficient in qE alone, qE and Z synthesis, or in qE, Z synthesis and also L synthesis were able to grow at 1800 micromol photons m(-2) s(-1) and exhibited no major symptoms of photooxidative stress. The mutants (and wild type) acclimated to HL by increasing photosynthetic capacity and decreasing light harvesting, which together rendered qE less important for photoprotection. At a metabolite level, the HL-grown mutants appeared to compensate for their remaining qE deficit with increased alpha-tocopherol and ascorbate levels compared to the wild type. The specificity of this response provides insight into the relationship between qE and the antioxidant network in plants.

Adaptation, Physiological↗

Physiological adjustments and arteriolar remodelling within skeletal muscle during acclimation to chronic hypoxia in the rat.

1. We have investigated the physiological and structural changes that occur in skeletal muscle vasculature during acclimation to chronic hypoxia in rats exposed to 12 % O2 in a hypoxic chamber for 7 or 18 days (7CH and 18CH rats, respectively) and in age-matched normoxic (7N and 18N) rats. 2. Under anaesthesia and breathing 12 % O2, 7CH and 18CH rats had lower arterial blood pressure (ABP) than 7N and 18N rats breathing air, but the haematocrit of the CH rats was increased so that their arterial O2 content equalled that of N rats. Blood flow recorded from the iliac or femoral artery and used to compute muscle vascular conductance (MVC: blood flow/ABP) showed that, in 18CH rats, MVC was comparable with that of 18N rats. 3. Maximal MVC induced by infusion of sodium nitroprusside (SNP) was used as an index of structural vascular conductance and compared with the MVC evoked by acute hypoxia (breathing 8 % O2). Hypoxia induced similar increases in MVC in 7N and 7CH rats and in 18N and 18CH rats, even though N rats were switched from air to 8 % O2 and CH rats were switched from 12 to 8 % O2. The MVCs attained with 8 % O2 and SNP were similar in 7N and 18N rats. However, the MVCs attained with 8 % O2 in 7CH and 18CH rats were only approximately 60 % of those evoked by SNP, while the MVC attained with SNP was greater in 18CH than in 18N rats. 4. Vascular casts of the spinotrapezius muscle analysed ex vivo showed that interbranch intervals along primary, secondary and terminal arterioles (22-50, 13-18 and 7-13 microm diameter, respectively) were 30-50 % shorter in 7CH and 18CH rats than in 7N and 18N rats. Further, the proportions of branches that were of the secondary and terminal arteriolar categories were increased such that the mean diameter of the branches was lower in 7CH than in 7N rats and lower in 18CH than in 18N rats. 5. These results indicate that arteriolar remodelling and angiogenesis occurs in skeletal muscle during acclimation to chronic hypoxia, beginning by the 7th day and progressing at least until the 18th day, so that the number of small arterioles and the functional size of the vascular bed is increased. We propose that these structural and functional changes enhance the ability of skeletal muscle to respond to acute hypoxia by facilitating the increase in vascular conductance, blood flow and thereby the O2 that can be delivered to muscle.

Adaptation, Physiological↗

Respiration of the external mycelium in the arbuscular mycorrhizal symbiosis shows strong dependence on recent photosynthates and acclimation to temperature.

* Although arbuscular mycorrhizal (AM) fungi are a major pathway in the global carbon cycle, their basic biology and, in particular, their respiratory response to temperature remain obscure. * A pulse label of the stable isotope (13)C was applied to Plantago lanceolata, either uninoculated or inoculated with the AM fungus Glomus mosseae. The extra-radical mycelium (ERM) of the fungus was allowed to grow into a separate hyphal compartment excluding roots. We determined the carbon costs of the ERM and tested for a direct temperature effect on its respiration by measuring total carbon and the (13)C:(12)C ratio of respired CO(2). With a second pulse we tested for acclimation of ERM respiration after 2 wk of soil warming. * Root colonization remained unchanged between the two pulses but warming the hyphal compartment increased ERM length. delta(13)C signals peaked within the first 10 h and were higher in mycorrhizal treatments. The concentration of CO(2) in the gas samples fluctuated diurnally and was highest in the mycorrhizal treatments but was unaffected by temperature. Heating increased ERM respiration only after the first pulse and reduced specific ERM respiration rates after the second pulse; however, both pulses strongly depended on radiation flux. * The results indicate a fast ERM acclimation to temperature, and that light is the key factor controlling carbon allocation to the fungus.

Acclimatization↗

Heat acclimation and heat stress have different effects on cholinergic muscarinic receptors.

In this brief report we have proved that membranal changes take place upon short-term heat acclimation. These changes may switch on a cascade of transient acclimatory compensatory responses as well as long-term processes. The changes observed in the MR profile are heat acclimation specific, and differ from those observed upon heat stress. These bring about functional changes in the signal transduction pathway for water secretion in the submaxillary salivary gland. A recovery period from heat stress, similar to STHA, leads to MR upregulation and decreased binding affinity. This response, however, is more pronounced than that observed upon STHA. The functional importance of the phenomenon is unclear. However, it could designate a novel component of the heat shock response.

Acclimatization↗