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D R Ort

Publications and source records attributed to D R Ort.

At least 19 recordsLinked to original sources
PubMed2006

Free-air carbon dioxide enrichment of soybean: influence of crop variety on residue decomposition.

Elevated atmospheric CO2 can result in larger plants returning greater amounts of residue to the soil. However, the effects of elevated CO2 on carbon (C) and nitrogen (N) cycling for different soybean varieties have not been examined. Aboveground residue of eight soybean [Glycine max (L.) Merr.] varieties was collected from a field study where crops had been grown under two different atmospheric CO2 levels [370 micromol mol(-1) (ambient) and 550 micromol mol(-1) (free-air carbon dioxide enrichment, FACE)]. Senesced residue material was used in a 60-d laboratory incubation study to evaluate potential C and N mineralization. In addition to assessing the overall effects of CO2 level and variety, a few specific variety comparisons were also made. Across varieties, overall residue N concentration was increased by FACE, but residue C concentration was only slightly increased. Overall residue C to N ratio was lower under FACE and total mineralized N was increased by FACE, suggesting that increased N2 fixation impacted residue decomposition; total mineralized C was also slightly increased by FACE. Across CO2 levels, varietal differences were also observed with the oldest variety having the lowest residue N concentration and highest residue C to N ratio; mineralized N was lowest in the oldest variety, illustrating the influence of high residue C to N ratio. It appears (based on our few specific varietal comparisons) that the breeding selection process may have resulted in some varietal differences in residue quality which can result in increased N or C mineralization under elevated CO2 conditions. This limited number of varietal comparisons indicated that more work investigating varietal influences on soil C and N cycling under elevated CO2 conditions is required.

Agriculture↗
PubMed2001

Impacts of chilling temperatures on photosynthesis in warm-climate plants.

Photosynthesis in warm-climate plants is substantially reduced after chilling. Tropical and subtropical species offer the opportunity to study the effects of low temperature on photosynthetic processes undisguised by the myriad of protective responses observed in temperate species. In this article, we highlight the primary components of photosynthesis that are affected by a short chill, in both the dark and the light, and discuss what is known of the mechanisms involved. Recent work implicates impaired redox and circadian regulation among other processes.

Carbohydrate Metabolism↗
PubMed2000

Differential effects of chilling-induced photooxidation on the redox regulation of photosynthetic enzymes.

Photosynthesis in plant species that are evolutionarily adapted for growth in warm climates is highly sensitive to illumination under cool conditions. Although it is well documented that illumination of these sensitive species under cool conditions results in the photosynthetic production of reactive oxygen molecules, the underlying mechanism for the inhibition of photosynthesis remains uncertain. Determinations of chloroplast fructose-1,6-bisphosphatase and sedoheptulose-1,7-bisphosphatase activity showed that the light-dependent, reductive activation of these key carbon reduction cycle enzymes was substantially inhibited in tomato (Lycopersicon esculentum) following illumination at 4 degrees C. However, other chloroplast enzymes also dependent on thioredoxin-mediated reductive activation were largely unaffected. We performed equilibrium redox titrations to investigate the thermodynamics of the thiol/disulfide exchange between thioredoxin f and the regulatory sulfhydryl groups of fructose-1,6-bisphosphatase, sedoheptulose-1,7-bisphosphatase, phosphoribulokinase, NADP-glyceraldehyde phosphate dehydrogenase, and the chloroplast ATPsynthase. We determined that the redox midpoint potentials for the regulatory sulfhydryl groups of the various enzymes spanned a broad range ( approximately 50 mV at pH 7. 9). The electron-sharing equilibria among thioredoxin f and its target enzymes largely explained the differential effects of photooxidation induced at low temperature on thioredoxin-mediated activation of chloroplast enzymes in tomato. These results not only provide a plausible mechanism for the low-temperature-induced inhibition of photosynthesis in this important group of plants, but also provide a quantitative basis to evaluate the influence of thioredoxin/target enzyme electron-sharing equilibria on the differential activation and deactivation kinetics of thioredoxin-regulated chloroplast enzymes.

ATP Synthetase Complexes↗
PubMed2000

An overnight chill induces a delayed inhibition of photosynthesis at midday in mango (Mangifera indica L.).

The effect of a cold night on photosynthesis in herbaceous chilling-sensitive crops, like tomato, has been extensively studied and is well characterized. This investigation examined the behaviour of the sub-tropical fruit tree, mango, to enable comparison with these well-studied systems. Unlike tomato, chilling between 5 degrees C and 7 degrees C overnight produced no significant inhibition of light-saturated CO(2) assimilation (A:) during the first hours following rewarming, measured either under controlled environment conditions or in the field. By midday, however, there was a substantial decline in A:, which could not be attributed to photoinhibition of PSII, but rather was associated with an increase in stomatal limitation of A: and lower Rubisco activity. Overnight chilling of tomato can cause severe disruption in the circadian regulation of key photosynthetic enzymes and is considered to be a major factor underlying the dysfunction of photosynthesis in chilling-sensitive herbaceous plants. Examination of the gas exchange of mango leaves maintained under constant conditions for 2 d, demonstrated that large depressions in A: during the subjective night were primarily the result of stomatal closure. Chilling did not disrupt the ability of mango leaves to produce a circadian rhythm in stomatal conductance. Rather, the midday increase in stomatal limitation of A: appeared to be the result of altered guard cell sensitivity to CO(2) following the dark chill.

Circadian Rhythm↗
PubMed1993

In vivo pulse labeling of proteins in attached leaves with radioactive amino acids.

The pulse labeling of proteins in attached leaves by the incorporation of radioactive amino acids can be a valuable approach to study a wide range of topics pertaining to plant gene expression. Transient changes in the rate of net protein synthesis can be measured since pulse labeling permits discrimination between proteins actively synthesized during the labeling interval from those accumulated at earlier times. Specific procedures for a rapid and convenient method of in vivo pulse labeling are outlined which we believe could be broadly useful in plant biological research. A particularly important technical feature of the protocol is electroblotting of radioactive proteins from the sodium dodecyl sulfate acrylamide resolving gel matrix to polyvinylidene difluoride transfer membrane. This step permits a single polyacrylamide slab gel to be used for autoradiography, immunoblot analysis, and protein staining, thereby greatly facilitating comparison among these complementary techniques and resulting in a significant savings in time and radioactive sample. Central considerations concerning the quantitation and interpretation of in vivo pulse labeling data are discussed.

Amino Acids↗
PubMed1992

Low temperature interrupts circadian regulation of transcriptional activity in chilling-sensitive plants.

Impaired chloroplast function is responsible for nearly two-thirds of the inhibition to net photosynthesis caused by dark chilling in tomato (Lycopersicon esculentum Mill.), yet it has not been possible to localize the dysfunction to specific chloroplast reactions. We report here on an effect that low-temperature exposure has in tomato on the expression of certain nuclear-encoded chloroplast proteins, which may be directly related to the chilling sensitivity of photosynthesis. Transcriptional activity of genes for both the chlorophyll a/b binding protein of photosystem II (Cab) as well as for ribulose 1,5-bisphosphate carboxylase/oxygenase (Rubisco) activase was found to be controlled by an endogenous rhythm. For Cab this rhythm was also visible at the level of newly synthesized protein, indicating that the circadian control of transcriptional activity normally ensures that this protein is synthesized only during daylight hours. However, low-temperature treatment suspended the timing of the rhythm in tomato so that, upon rewarming, the circadian control was reestablished but was displaced from the actual time of day by the length of the chilling exposure. In addition, we found that the normal turnover of Cab and Rubisco activase mRNA was suspended during the low-temperature treatment, but, upon rewarming, this stabilized message was not translated into protein. We believe that the low-temperature-induced mistiming of gene expression together with its effect on the translatability of existing transcripts may be an important clue in unraveling the basis for the chilling sensitivity of photosynthesis in tomato.

Chloroplasts↗
PubMed1992

Spatial Distribution of Photosynthesis during Drought in Field-Grown and Acclimated and Nonacclimated Growth Chamber-Grown Cotton.

Inhomogeneous photosynthetic activity has been reported to occur in drought-stressed leaves. In addition, it has been suggested that these water stress-induced nonuniformities in photosynthesis are caused by "patchy" stomatal closure and that the phenomenon may have created the illusion of a nonstomatal component to the inhibition of photosynthesis. Because these earlier studies were performed with nonacclimated growth chamber-grown plants, we sought to determine whether such "patches" existed in drought-treated, field-grown plants or in chamber-grown plants that had been acclimated to low leaf water potentials (psi(leaf)). Cotton (Gossypium hirsutum L.) was grown in the field and subjected to drought by withholding irrigation and rain from 24 d after planting. The distribution of photosynthesis, which may reflect the stomatal aperture distribution in a heterobaric species such as cotton, was assayed by autoradiography after briefly exposing attached leaves of field-grown plants to (14)CO(2). A homogeneous distribution of radioactive photosynthate was evident even at the lowest psi(leaf) of -1.34 MPa. "Patchiness" could, however, be induced by uprooting the plant and allowing the shoot to air dry for 6 to 8 min. In parallel studies, growth chamber-grown plants were acclimated to drought by withholding irrigation for three 5-d drought cycles interspersed with irrigation. This drought acclimation lowered the psi(leaf) value at which control rates of photosynthesis could be sustained by approximately 0.7 MPa and was accompanied by a similar decline in the psi(leaf) at which patchiness first appeared. Photosynthetic inhomogeneities in chamber-grown plants that were visible during moderate water stress and ambient levels of CO(2) could be largely removed with elevated CO(2) levels (3000 muL L(-1)), suggesting that they were stomatal in nature. However, advanced dehydration (less than approximately 2.0 MPa) resulted in "patches" that could not be so removed and were probably caused by nonstomatal factors. The demonstration that patches do not exist in drought-treated, field-grown cotton and that the presence of patches in chamber-grown plants can be altered by treatments that cause an acclimation of photosynthesis leads us to conclude that spatial heterogeneities in photosynthesis probably do not occur frequently under natural drought conditions.

Journal Article↗
PubMed1991

Photophosphorylation in Attached Leaves of Helianthus annuus at Low Water Potentials.

The in situ response of photophosphorylation and coupling factor activity to low leaf water potential (psi(L)) was investigated using kinetic spectroscopy to measure the flash-induced electrochromic absorption change in attached sunflower (Helianthus annuus L. cv IS894) leaves. The electrochromic change is caused by the formation of an electric potential across the thylakoid membrane associated with proton uptake. Since depolarization of the thylakoid membrane following flash excitation is normally dominated by proton efflux through the coupling factor during ATP formation, this measurement can provide direct information about the catalytic activity of the coupling factor. Under low psi(L) conditions in which a clear nonstomatal limitation of net photosynthesis could be demonstrated, we found a strong inhibition of coupling factor activity in dark-adapted leaves which was probably caused by an increase in the energetic threshold for the activation of the enzyme at low psi(L). While this result supported earlier in vitro findings, we further discovered that the light-dependent reduction of coupling factor reversed any observable effect of low psi(L) on the energetics of activation or on photophosphorylation competence. Furthermore, coupling factor was reduced, even in severely droughted sunflower, almost immediately upon illumination. Based on these measurements, we conclude that the nonstomatal limitation of photosynthesis observed by us and others in droughted plants cannot be explained by impaired coupling factor activity.

Journal Article↗
PubMed1990

Impaired reductive activation of stromal bisphosphatases in tomato leaves following low-temperature exposure at high light.

Photosynthesis in domestic tomato (Lycopersicon esculentum L.) is highly sensitive to low temperature, particularly when accompanied by high light. Since previous studies have established that the inhibited plants retain photosynthetic electron transfer and ATP formation competence, we sought to identify specific steps in the photosynthetic carbon reduction pathway that could account for the lost photosynthetic capacity. Measurements of steady-state photosynthetic metabolite pool sizes showed an accumulation of fructose 1,6-bisphosphate and sedohepulose 1,7-bisphosphate following chilling in the light. Measurements of in vivo turnover rates of the metabolite pools accompanied by direct determinations of enzymatic activity showed that the capacity of the stromal bisphosphatases was substantially reduced following chilling in the light and was the cause of the bisphosphate accumulation. The time course of the loss of phosphatase activity closely mimicked that of the inhibition of net photosynthesis, further indicating that impaired phosphatase function is the underlying cause of the sensitivity of photosynthesis in tomato to light and chilling. Fructose 1,6-bisphosphatase extracted from inhibited tomato plants could be fully activated in the presence of dithiothreitol, indicating that chilling in the light disrupts the normal, thioredoxin-dependent, activation pathway of the stromal bisphosphatases. This disruption could involve a change in the redox potential of the functional disulfide on the phosphatases.

Cold Temperature↗
PubMed1989

Photophosphorylation after chilling in the light : effects on membrane energization and coupling factor activity.

The response of in situ photophosphorylation in attached cucumber (Cucumis sativus L. cv Ashley) leaves to chilling under strong illumination was investigated. A single-beam kinetic spectrophotometer fitted with a clamp-on, whole leaf cuvette was used to measure the flash-induced electrochromic absorbance change at 518 minus 540 nanometers (DeltaA(518-540)) in attached leaves. The relaxation kinetics of the electric field-indicating DeltaA(518-540) measures the rate of depolarization of the thylakoid membrane. Since this depolarization process is normally dominated by proton efflux through the coupling factor during ATP synthesis, this technique can be used, in conjuction with careful controls, as a monitor of in situ ATP formation competence. Whole, attached leaves were chilled at 5 degrees C and 1000 microeinsteins per square meter per second for up to 6 hours then rewarmed in the dark at room temperature for 30 minutes and 100% relative humidity. Leaf water potential, chlorophyll content, and the effective optical pathlength for the absorption measurements were not affected by the treatment. Light- and CO(2)-saturated leaf disc oxygen evolution and the quantum efficiency of photosynthesis were inhibited by approximately 50% after 3 hours of light chilling and by approximately 75% after 6 hours. Despite the large inhibition to net photosynthesis, the measurements of DeltaA(518-540) relaxation kinetics showed photophosphorylation to be largely unaffected by the chilling and light exposure. The amplitude of the DeltaA(518-540) measures the degree of energization of the photosynthetic membranes and was reduced significantly by chilling in the light. The cause of the decreased energization was traced to impaired turnover of photosystem II. Our measurements showed that the chilling of whole leaves in the light caused neither an uncoupling of photophosphorylation from photosynthetic electron transport nor any irreversible inhibition of the chloroplast coupling factor in situ. The sizeable inhibition in net photosynthesis observed after chilling in the light cannot, therefore, be attributed to any direct effect on photophosphorylation competence.

Journal Article↗
PubMed1988

Changes in protein synthesis induced in tomato by chilling.

Impaired chloroplast function is responsible for nearly two-thirds of the inhibition of net photosynthesis caused by dark chilling in tomato (Lycopersicon esculentum Mill.). Yet the plant can eventually recover full photosynthetic capacity if it is rewarmed in darkness at high relative humidity. As a means of identifying potential sites of chilling injury in tomato, we monitored leaf protein synthesis in chilled plants during this rewarming recovery phase, since changes in the synthesis of certain proteins might be indicative of damaged processes in need of repair. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of proteins pulse labeled with [(35)S]methionine revealed discrete changes in the pattern of protein synthesis as a result of chilling. A protein of M(r) = 27 kilodaltons (kD), abundantly synthesized by unchilled plants, declined to undetectable levels in chilled plants. Reillumination restored the synthesis of this protein in plants rewarmed for 8 hours. Peptide mapping analysis showed the 27 kD protein to be the major chlorophyll a/b binding protein of the photosystem II light-harvesting complex (LHCP-II). The identity of this protein was confirmed by its immunoprecipitation from leaf extracts by a monoclonal antibody specific for the major LHCP-II species. While chilling abolished the synthesis of the major LHCP-II species, it also induced the synthesis of an entirely new protein of M(r) = 35 kD. The protein was synthesized on cytoplasmic ribosomes, and two-dimensional polyacrylamide gel electrophroesis showed it to exist as a single isoelectric species. This chilling-induced 35 kD protein is structurally distinct from the 27 kD LHCP-II and appears to be synthesized specifically in response to low temperature. While the 35 kD protein was found not to be associated with the chloroplast thylakoid membrane, chilling did cause selective changes in thylakoid membrane protein synthesis. The synthesis of two unidentified proteins, M(r) = 14 and 41 kD, and the beta-subunit of the chloroplast coupling factor were substantially reduced after chilling. These losses may provide clues as to the causes of the overall reduction in net photosynthesis caused by chilling.

Journal Article↗
PubMed1988

Photosynthesis and temperature, with particular reference to effects on quantum yield.

Previous reviews of the effects of temperature on in vivo photosynthesis have mainly concerned the effects of temperature on light saturated rates. The quantum yield of photosynthesis (phi), as a measure of light limited photosynthesis, has generally been regarded as temperature insensitive. At temperatures close to the minima and maxima at which plants can sustain photosynthetic CO2 assimilation, light may damage the photosynthetic apparatus, an effect termed photoinhibition. A constant feature of photoinhibition is a reduction in phi. In maize, chilling-dependent photoinhibition reduces both phi and the light saturated rate of CO2 assimilation (Asat) and of O2 evolution. Analysis of recovery of CO2 uptake in these leaves suggests that whilst Asat recovers in a few hours, phi may not be fully restored for days. Examination of mature crop canopies shows that only a small proportion of the leaves are likely to become light saturated and then only for part of the day. The relative significance of temperature-induced changes in Asat and phi have also been tested in canopy models of maize crop photosynthesis. These suggest that whilst changes in either parameter will have similar effects on total canopy photosynthesis on the sunniest days of the year, for an average summer's day changes in phi will be of far greater importance. Consideration is therefore given to the factors associated with thylakoid membranes that may determine temperature-induced decreases in phi. Chilling of maize leaves under high light levels reduces the quantum yield of PSII and whole chain electron transport in concert with a decrease in the capacity of isolated thylakoids to bind atrazine, which is indicative of a loss or damage to the QB protein. Besides such classical symptoms of photoinhibition of PSII, chilling also induces the accumulation of a 31 kDa polypeptide in the thylakoids of maize leaves. This polypeptide fractionates with the light-harvesting chlorophyll a/b protein complex (LHCII) and has been tentatively identified as an unprocessed precursor of CP29 since it binds chlorophyll and is immunologically related to CP29. Accumulation of the 31 kDa polypeptide is associated with a modification in the energetics of LHCII, which may result in a decrease in excitation energy from LHCII to PSII and contribute to a decrease in phi. Examination is also made of how stress-induced modifications of interactions between PSII complexes, functioning of the cyt b6/f complex, the permeability of the thylakoid membrane to protons and the activity of the coupling factor may contribute to decreases in phi.

Cold Temperature↗
PubMed1987

The effects of chloroplast coupling factor reduction on the energetics of activation and on the energetics and efficiency of ATP formation.

There are two separate aspects to the energetic control over the initiation of ATP formation by chloroplast thylakoid membranes: (i) the free energy requirement for the phosphorylation of ADP and (ii) the energy-dependent transformation of the coupling factor into a catalytically active state. Using flash excitation, we have been able to distinguish between limitations imposed on ATP formation by the energetics of coupling factor activation from limitations imposed by the thermodynamics of ADP phosphorylation. We have placed particular emphasis on the effects of coupling factor reduction. ATP hydrolysis, ATP synthesis, and the release of tightly bound [14C]ADP were initiated by fewer flashes in thylakoid membranes in which the coupling factors were prereduced than in membrane samples in which the coupling factor remained oxidized. Only under conditions in which ATP synthesis occurred against a large chemical potential (i.e. delta GATP) did the onset of net ATP synthesis require a larger electrochemical potential than did the activation of the coupling factor. Thus, only at delta GATP values greater than about 51 kJ.mol-1 for thylakoid membranes with oxidized coupling factors and about 45 kJ.mol-1 when the coupling factors are reduced was the onset of ATP synthesis dictated by the thermodynamic equilibrium between ATP and ADP. At lower delta GATP values, the electrochemical potential became energetically adequate to phosphorylate ADP prior to becoming adequate to activate the coupling factor. These data indicate that reduction of the coupling factor lowers the delta pH necessary for activation from approximately 2.9 to approximately 2.6 units. Accompanying the lower energetic requirement for the activation of the reduced coupling factor was an increase in the efficiency of ATP synthesis under conditions of limiting energization.

Adenosine Triphosphate↗
PubMed1986

Quantitation of 2,5-dibromo-3-methyl-6-isopropyl-p-benzoquinone binding sites in chloroplast membranes: evidence for a functional dimer of the cytochrome b6f complex.

The binding of 2,5-dibromo-3-methyl-6-isopropyl-p-benzoquinone (DBMIB) to chloroplast thylakoid membranes was investigated by analyzing the inhibition of electron transfer by DBMIB according to a steady-state rate relationship for enzyme-catalyzed reactions in the presence of tightly binding reversible inhibitors. DBMIB interacts with the cytochrome b6f complex in a manner best described by an apparent dissociation constant near 6 nM. The binding site titer is 1 mmol X mol chlorophyll-1. This number of DBMIB binding sites approaches one-half the number of cytochrome b6f complexes present in the membrane. These data suggest that the cytochrome b6f complex may function in electron transfer as a dimer, plastoquinol oxidation being totally inhibited by the binding of a single DBMIB molecule to the dimer.

Chloroplasts↗