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Relationship of Electrical Conductance at Two Frequencies to Cold Injury and Acclimation in Cornus stolonifera Michx.

The ratio of electrical conductance measured at two frequencies can be used to predict the cold hardiness of stem sections of Cornus stolonifera Michx. during the first stage of cold acclimation. Electrical conductance at 50 hertz divided by electrical conductance at 100 kilohertz gave a better estimate of hardiness than measurements at either frequency alone. The observed increase in the electrical conductance ratio as hardiness increased is consistent with an increase in membrane permeability. After plants were exposed to nonlethal frost, hardiness increased rapidly, and the relation between the conductance ratio and hardiness changed. This change indicates that ice crystallization induces a significant physiological alteration in the plants. Contrary to expectations, stem sections exposed to lethal temperatures could not consistently be separated from sections exposed to nonlethal temperatures by electrical conductance ratio measurements made immediately after thawing.

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Nucleic Acid and protein changes in relation to cold acclimation and freezing injury of korean boxwood leaves.

Quantitative and qualitative differences in nucleic acids of Korean boxwood (Buxus microphylla var. Koreana) leaves were determined by methylated albumin kieselguhr chromatography at different levels of cold hardiness. During cold acclimation there was an increase in RNA, mainly ribosomal RNA, with little or no change in DNA. The increase in ribosomal RNA was closely paralleled by an increase in water soluble and membrane bound proteins. As cold hardiness increased, ribonuclease activity declined.Exposure of hardy boxwood plants to warm temperatures resulted in a rapid loss in cold resistance and a rapid synthesis of nucleic acids as judged by (32)P incorporation.Following a killing frost to Korean boxwood leaves, there was a rapid decrease in all nucleic acid fractions which was attributed to nuclease activity. Within 5 hours there was no measurable soluble RNA and ribosomal RNA. Tenaciously bound RNA was somewhat more persistent.

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Photosynthetic Acclimation to Temperature in the Desert Shrub, Larrea divaricata: II. Light-harvesting Efficiency and Electron Transport.

The response of photosynthetic electron transport and light-harvesting efficiency to high temperatures was studied in the desert shrub Larrea divaricata Cav. Plants were grown at day/night temperatures of 20/15, 32/25, or 45/33 C in rough approximation of natural seasonal temperature variations. The process of acclimation to high temperatures involves an enhancement of the stability of the interactions between the light-harvesting pigments and the photosystem reaction centers. As temperature is increased, the heat-induced dissociation of these complexes results in a decrease in the quantum yield of electron transport at limiting light intensity, followed by a loss of electron transport activity at rate-saturating light intensity. The decreased quantum yield can be attributed to a block of excitation energy transfer from chlorophyll b to chlorophyll a, and changes in the distribution of the excitation energy between photosystems II and I. The block of excitation energy transfer is characterized by a loss of the effectiveness of 480 nm light (absorbed primarily by chlorophyll b) to drive protochemical processes, as well as fluorescence emission by chlorophyll b.

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Photosynthate partitioning in soybean leaves at two irradiance levels: comparative responses of acclimated and unacclimated leaves.

High irradiance-acclimated soybean leaves had the same CO(2) exchange rates, but lower starch accumulation rates and correspondingly higher translocation rates than unacclimated leaves. Increased translocation rates were associated with increased sucrose phosphate synthetase (EC 2.4.1.14) activity. Foliar sucrose levels and adenosine diphosphate-glucose pyrophosphorylase (EC 2.7.7.9) activity were unaffected. Carbon assimilation, partitioning, and enzyme activity of unacclimated leaves were unaltered even after a second day's exposure to high irradiance. Results are consistent with the hypothesis that photosynthate partitioning between starch synthesis and sucrose translocation are controlled in part by the rate of sucrose synthesis.

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Interaction between External and Internal Conditions in the Development of Photosynthetic Features in a Grass Leaf: I. REGIONAL RESPONSES ALONG A LEAF DURING AND AFTER LOW-LIGHT OR HIGH-LIGHT ACCLIMATION.

Morphological and functional features were compared along a developing third leaf and fully expanded leaf from high-light- and low-light-acclimated seedlings of Lolium multiflorum.The young leaf contains a gradient of differentiating tissue, ranging from meristematic cells at the leaf base to mature tissue at the tip; this gradient can be related to the maturation of a functional photosynthetic apparatus. Along the fully expanded leaf, a decreasing gradient from tip to base is maintained for functional characteristics (net maximum photosynthesis, chlorophyll content, and ribulose bisphosphate carboxylase activity) and for a number of structural parameters (number of mesophyll cells and their external surface area, number of chloroplasts and their envelope area), irrespective of the light regime. In contrast, a constancy in the absolute intrachloroplastic lamellar content per plastid was revealed whatever the position in the leaf or irradiance received. However, the relative membrane content was lower in high-light chloroplasts due to their larger volume compared to low-light plastids (dilution effect).The longitudinal differences in functional and morphological characteristics are interpreted as the result of interaction between the internal gradient of differentiating tissue along a developing young leaf and the external light conditions during development.

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Structural Changes in Thylakoid Proteins during Cold Acclimation and Freezing of Winter Rye (Secale cereale L. cv. Puma).

Thylakoids were isolated from nonhardened and cold-hardened winter rye (Secale cereale L. cv. Puma), and subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis in the presence and absence of sulfhydryl reagents. Electrophoresis of cold-hardened rye thylakoid proteins revealed the presence of a 35 kilodalton polypeptide and the absence of a 51 kilodalton polypeptide found in nonhardened rye thylakoid proteins. The 35 kilodalton band could be induced by adding beta-mercaptoethanol to nonhardened rye thylakoid proteins, whereas the 51 kilodalton band could be formed by adding cupric phenanthroline to these same proteins. Sulfhydryl group titration showed that cold-hardened rye thylakoid proteins contained more free sulfhydryls than nonhardened rye proteins. Although amino acid analysis of thylakoid proteins revealed quantitative differences in several amino acid residues, the polarity of thylakoid proteins did not change during cold acclimation. No significant changes in sodium dodecyl sulfate-polyacrylamide gels of thylakoid proteins appeared when either nonhardened or cold-hardened plants were frozen in vivo or in vitro. However, thylakoid proteins did aggregate when frozen in the presence of beta-mercaptoethanol. Although thylakoid proteins isolated from cold-hardened rye contained more reduced thiols, a general state of reduction did not act as a cryoprotectant. It is hypothesized that conformational changes of specific proteins may be important for low temperature growth of rye.

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Acclimation to High CO(2) in Bean : Carbonic Anhydrase and Ribulose Bisphosphate Carboxylase.

Young bean plants (Phaseolus vulgaris L. cv Seafarer) grew faster in air enriched with CO(2) (1200 microliters per liter) than in ambient CO(2) (330 microliters per liter). However, by 7 days when increases in overall growth (dry weight, leaf area) were visible, there was a significant decline (about 25%) in the leaf mineral content (N, P, K, Ca, Mg) and a drop in the activity of two enzymes of carbon fixation, carbonic anhydrase and ribulose 1,5-bisphosphate (RuBP) carboxylase under high CO(2). Although the activity of neither enzyme was altered in young, expanding leaves during the acclimation period, in mature leaves the activity of carbonic anhydrase was reduced 95% compared with a decline of 50% in ambient CO(2). The drop in RuBP carboxylase was less extreme with 40% of the initial activity retained in the high CO(2) compared with 50% in the ambient atmosphere. While CO(2) enrichment might alter the flow of carbon into the glycolate pathway by modifying the activities of carbonic anhydrase or RuBP carboxylase, there is no early change in the ability of photosynthetic tissue to oxidize glycolate to CO(2).

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Acclimation Processes in the Light-Harvesting System of the Cyanobacterium Anacystis nidulans following a Light Shift from White to Red Light.

Cyanobacteria acclimate to changes in light by adjusting the amounts of different cellular compounds, for example the light-harvesting macromolecular complex. Described are the acclimatization responses in the light-harvesting system of the cyanobacterium Anacystis nidulans following a shift from high intensity, white light to low intensity, red light.The phycocyanin and chlorophyll content and the relative amount of the two linker peptides (33 and 30 kilodaltons) in the phycobilisome were studied. Both the phycocyanin and chlorophyll content per cell increased after the shift, although the phycocyanin increased relatively more. The increase in phycocyanin was biphasic in nature, a fast initial phase and a slower second phase, while the chlorophyll increase was completed in one phase. The phycocyanin and chlorophyll responses to red light were immediate and were completed within 30 and 80 hours for chlorophyll and phycocyanin, respectively. An immediate response was also seen for the two phycobilisome linker peptides. The amount of both of them increased after the shift, although the 33 kilodalton linker peptide increased faster than the 30 kilodalton linker peptide. The increase of the content of the two linker peptides stopped when the phycocyanin increase shifted from the first to the second phase. We believe that the first phase of phycocyanin increase was due mainly to an increase in the phycobilisome size while the second phase was caused only by an increase in the amount of phycobilisomes. The termination of chlorophyll accumulation, which indicates that no further reaction center chlorophyll antennae were formed, occurred parallel to the onset of the second phase of phycocyanin accumulation.

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Primary Role of the Cytoplasmic Membrane in Thermal Acclimation Evidenced in Nitrate-Starved Cells of the Blue-Green Alga, Anacystis nidulans.

The lipid phase transition of the cytoplasmic membrane and the chilling susceptibility were studied in nitrate-starved Anacystis nidulans cells. Nitrate starvation resulted in the disappearance of the thylakoid membrane system, without any effect on chilling susceptibility. The chilling susceptibility of the algal cells depended on the growth temperature. Temperatures of lipid phase transitions of the cytoplasmic membranes were detected by chilling-induced spectral changes in the carotenoid region, in vivo. These values were identical to those of cultures containing intact thylakoid systems. Our results suggest that cytoplasmic membrane plays a determinative role in the thermal acclimation of the alga cells.

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Acclimation of Ribulose Bisphosphate Carboxylase and mRNAs to Changing Irradiance in Adult Tobacco Leaves: Differential Expression in LSU And SSU mRNA.

The transfer of Nicotiana tabacum plants grown in low light (60 micromoles quanta per square meter per second) to higher light (360 micromoles quanta per square meter per second) was previously shown to induce adaptive stimulation of photosynthetic capacities. The variations of ribulose bisphosphate carboxylase/oxygenase (RubisCo) expression in mature leaves was examined as a result of this acclimation. Maximum or initial activities increased markedly after low- to high-light transfer with a maximum effect after 2 to 3 days. The higher activity is mainly explained by RubisCo protein synthesis as shown by immunorocket technique. Small subunits of RubisCo (SSU) mRNA relative content determined by hybridization of total RNA with DNA probe by Dot-blot method, followed the same pattern as RubisCo quantity. The magnitude of this response was amplified when more contrasting light conditions (25 versus 360 micromoles per square meter per second) were established on the same leaf: RubisCo activity, RubisCo protein, and SSU mRNA contents decreased in the shaded zone and increased in the high-light zone within 1 day. After 2 days the shade/light ratio was 1 to 3 for RubisCo protein and 1 to 4 for SSU-RNA, whereas the ratios remained equal to one in controls. Hybridization of the same RNA extracts with large subunits of RubisCo (LSU) probe showed no variation in LSU-RNA content. So in green adult leaves, the expression of SSU and LSU genes is regulated differently. The observed white light quantitative effect on RubisCo expression was not dependent on the photosynthetic rate or assimilate content since low CO(2) concentration around the leaf after the light shift did not modify the response.

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Influence of Drought Acclimation and CO(2) Enrichment on Osmotic Adjustment and Chlorophyll a Fluorescence of Sunflower during Drought.

Osmotic adjustment occurred during drought in expanded leaves of sunflowers (Helianthus annuus var Hysun 30) which had been continuously exposed to 660 microliters CO(2) per liter or had been previously acclimated to drought. The effect was greatest when the treatments were combined and was negligible in nonacclimated plants grown at 340 microliters CO(2) per liter. The concentrations of ethanol soluble sugars and potassium increased during drought but they did not account for the osmotic adjustment. The delay in the decline in conductance and relative water content and in the loss of structural integrity with increasing drought was dependent on the degree of osmotic adjustment. Where it was greatest, conductance fell from 5.8 millimeters per second on the first day of drought to 1.3 millimeters per second on the fourth day and was at approximately the same level on the eighth day. The relative water content remained constant at 85% for three days and fell to 36% on the sixth day. There was no evidence of leaf desiccation even on the eighth day. In contrast, the conductance of leaves showing minimal adjustment fell rapidly after the first day of drought and was negligible after the fourth, at which time the relative water content was 36%. By the sixth day of drought, areas near the margins of the leaves were desiccating and the plants did not recover upon rewatering. Despite the differences in the rate of change of conductance and relative water content during drought, photosynthetic electron transport activity, inferred from measurements of chlorophyll a fluorescence in vivo and PSII activity of isolated thylakoids, remained functional until desiccation occurred.

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Acclimation of Photosynthesis to Elevated CO(2) in Five C(3) Species.

The effect of long-term (weeks to months) CO(2) enhancement on (a) the gas-exchange characteristics, (b) the content and activation state of ribulose-1,5-bisphosphate carboxylase (rubisco), and (c) leaf nitrogen, chlorophyll, and dry weight per area were studied in five C(3) species (Chenopodium album, Phaseolus vulgaris, Solanum tuberosum, Solanum melongena, and Brassica oleracea) grown at CO(2) partial pressures of 300 or 900 to 1000 microbars. Long-term exposure to elevated CO(2) affected the CO(2) response of photosynthesis in one of three ways: (a) the initial slope of the CO(2) response was unaffected, but the photosynthetic rate at high CO(2) increased (S. tuberosum); (b) the initial slope decreased but the CO(2)-saturated rate of photosynthesis was little affected (C. album, P. vulgaris); (c) both the initial slope and the CO(2)-saturated rate of photosynthesis decreased (B. oleracea, S. melongena). In all five species, growth at high CO(2) increased the extent to which photosynthesis was stimulated following a decrease in the partial pressure of O(2) or an increase in measurement CO(2) above 600 microbars. This stimulation indicates that a limitation on photosynthesis by the capacity to regenerate orthophosphate was reduced or absent after acclimation to high CO(2). Leaf nitrogen per area either increased (S. tuberosum, S. melongena) or was little changed by CO(2) enhancement. The content of rubisco was lower in only two of the five species, yet its activation state was 19% to 48% lower in all five species following long-term exposure to high CO(2). These results indicate that during growth in CO(2)-enriched air, leaf rubisco content remains in excess of that required to support the observed photosynthetic rates.

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Acclimation of Two Tomato Species to High Atmospheric CO(2): I. Sugar and Starch Concentrations.

Lycopersicon esculentum Mill. cv Vedettos and Lycopersicon chmielewskii Rick, LA 1028, were exposed to two CO(2) concentrations (330 or 900 microliters per liter) for 10 weeks. Tomato plants grown at 900 microliters per liter contained more starch and more sugars than the control. However, we found no significant accumulation of starch and sugars in the young leaves of L. esculentum exposed to high CO(2). Carbon exchange rates were significantly higher in CO(2)-enriched plants for the first few weeks of treatment but thereafter decreased as tomato plants acclimated to high atmospheric CO(2). This indicates that the long-term decline of photosynthetic efficiency of leaf 5 cannot be attributed to an accumulation of sugar and/or starch. The average concentration of starch in leaves 5 and 9 was always higher in L. esculentum than in L. chmielewskii (151.7% higher). A higher proportion of photosynthates was directed into starch for L. esculentum than for L. chmielewskii. However, these characteristics did not improve the long-term photosynthetic efficiency of L. chmielewskii grown at high CO(2) when compared with L. esculentum. The chloroplasts of tomato plants exposed to the higher CO(2) concentration exhibited a marked accumulation of starch. The results reported here suggest that starch and/or sugar accumulation under high CO(2) cannot entirely explain the loss of photosynthetic efficiency of high CO(2)-grown plants.

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Acclimation of Two Tomato Species to High Atmospheric CO(2): II. Ribulose-1,5-Bisphosphate Carboxylase/Oxygenase and Phosphoenolpyruvate Carboxylase.

Lycopersicon esculentum Mill. cv Vedettos and Lycopersicon chmielewskii Rick, LA 1028, were exposed to two CO(2) concentrations (330 or 900 microliters per liter) for 10 weeks. The elevated CO(2) concentrations increased the initial ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) activity of both species for the first 5 weeks of treatment but the difference did not persist during the last 5 weeks. The activity of Mg(2+)-CO(2)-activated Rubisco was higher in 900 microliters per liter for the first 2 weeks but declined sharply thereafter. After 10 weeks, leaves grown at 330 microliters per liter CO(2) had about twice the Rubisco activity compared with those grown at 900 microliters per liter CO(2). The two species showed the same trend to Rubisco declines under high CO(2) concentrations. The percent activation of Rubisco was always higher under high CO(2). The phosphoenolpyruvate carboxylase (PEPCase) activity measured in tomato leaves averaged 7.9% of the total Rubisco. PEPCase showed a similar trend with time as the initial Rubisco but with no significant difference between nonenriched and CO(2)-enriched plants. Long-term exposure of tomato plants to high CO(2) was previously shown to induce a decline of photosynthetic efficiency. Based on the current study and on previous results, we propose that the decline of activated Rubisco is the main cause of the acclimation of tomato plants to high CO(2) concentrations.

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Root restriction as a factor in photosynthetic acclimation of cotton seedlings grown in elevated carbon dioxide.

Interactive effects of root restriction and atmospheric CO(2) enrichment on plant growth, photosynthetic capacity, and carbohydrate partitioning were studied in cotton seedlings (Gossypium hirsutum L.) grown for 28 days in three atmospheric CO(2) partial pressures (270, 350, and 650 microbars) and two pot sizes (0.38 and 1.75 liters). Some plants were transplanted from small pots into large pots after 20 days. Reduction of root biomass resulting from growth in small pots was accompanied by decreased shoot biomass and leaf area. When root growth was less restricted, plants exposed to higher CO(2) partial pressures produced more shoot and root biomass than plants exposed to lower levels of CO(2). In small pots, whole plant biomass and leaf area of plants grown in 270 and 350 microbars of CO(2) were not significantly different. Plants grown in small pots in 650 microbars of CO(2) produced greater total biomass than plants grown in 350 microbars, but the dry weight gain was found to be primarily an accumulation of leaf starch. Reduced photosynthetic capacity of plants grown at elevated levels of CO(2) was clearly associated with inadequate rooting volume. Reductions in net photosynthesis were not associated with decreased stomatal conductance. Reduced carboxylation efficiency in response to CO(2) enrichment occurred only when root growth was restricted suggesting that ribulose-1,5-bisphosphate carboxylase/oxygenase activity may be responsive to plant source-sink balance rather than to CO(2) concentration as a single factor. When root-restricted plants were transplanted into large pots, carboxylation efficiency and ribulose-1,5-bisphosphate regeneration capacity increased indicating that acclimation of photosynthesis was reversible. Reductions in photosynthetic capacity as root growth was progressively restricted suggest sink-limited feedback inhibition as a possible mechanism for regulating net photosynthesis of plants grown in elevated CO(2).

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Sucrose Synthase Expression during Cold Acclimation in Wheat.

When wheat (Triticum aestivum) seedlings are exposed to a cold temperature (2-4 degrees C) above 0 degrees C, sucrose accumulates and sucrose synthase activity increases. The effect of a cold period on the level of sucrose synthase (SS) was investigated. Using antibodies against wheat germ SS, Western blots studies showed that the amount of the SS peptide increased during 14 days in the cold, when plants were moved from 23 degrees C to 4 degrees C. The level of SS diminished when plants were moved back to 23 degrees C. Northern blots of poly(A)(+) RNA, confirmed a five- to sixfold induction of SS in wheat leaves during cold acclimation. These results indicate that SS is involved in the plant response to a chilling stress.

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Enhancement of Anaerobic Respiration in Root Tips of Zea mays following Low-Oxygen (Hypoxic) Acclimation.

Root tips (10-millimeter length) were excised from hypoxically pretreated (HPT, 4% [v/v] oxygen at 25 degrees C for 16 hours) or nonhypoxically pretreated (NHPT, 40% [v/v] oxygen) maize (Zea mays) plants, and their rates of respiration were compared by respirometry under aerobic and anaerobic conditions with exogenous glucose. The respiratory quotient under aerobic conditions with 50 millimolar glucose was approximately 1.0, which is consistent with glucose or other hexose sugars being utilized as the predominant carbon source in glycolysis. Under strictly anaerobic conditions (anoxia), glycolysis was accelerated appreciably in both HPT and NHPT root tips, but the rate of anaerobic respiration quickly declined in NHPT roots. [U-(14)C]Glucose supplied under anaerobic conditions was taken up and respired by HPT root tips up to five times more rapidly than by NHPT roots. When anaerobic ethanol production was measured with excised root tips in 50 millimolar glucose, HPT tissues consistently produced ethanol more rapidly than NHPT tissues. These data suggest that a period of low oxygen partial pressure is necessary to permit adequate acclimation of the root tip of maize to subsequent anoxia, resulting in more rapid rates of fermentation and generation of ATP.

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Accumulation of an acidic dehydrin in the vicinity of the plasma membrane during cold acclimation of wheat

Expression of the acidic dehydrin gene wcor410 was found to be associated with the development of freezing tolerance in several Gramineae species. This gene is part of a family of three homologous members, wcor410, wcor410b, and wcor410c, that have been mapped to the long arms of the homologous group 6 chromosomes of hexaploid wheat. To gain insight into the function of this gene family, antibodies were raised against the WCOR410 protein and affinity purified to eliminate cross-reactivity with the WCS120 dehydrin-like protein of wheat. Protein gel blot analyses showed that the accumulation of WCOR410 proteins correlates well with the capacity of each cultivar to cold acclimate and develop freezing tolerance. Immunoelectron microscope analyses revealed that these proteins accumulate in the vicinity of the plasma membrane of cells in the sensitive vascular transition area where freeze-induced dehydration is likely to be more severe. Biochemical fractionation experiments indicated that WCOR410 is a peripheral protein and not an integral membrane protein. These results provide direct evidence that a subtype of the dehydrin family accumulates near the plasma membrane. The properties, abundance, and localization of these proteins suggest that they are involved in the cryoprotection of the plasma membrane against freezing or dehydration stress. We propose that WCOR410 plays a role in preventing the destabilization of the plasma membrane that occurs during dehydrative conditions.

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