Search PubMed⌕ Search

Biomedical subjects

G A Berkowitz

Publications and source records attributed to G A Berkowitz.

33 records · Page 2Linked to original sources
PubMed1990

Correlation between the Maintenance of Photosynthesis and in Situ Protoplast Volume at Low Water Potentials in Droughted Wheat.

Studies were undertaken to examine the relationship between water deficit effects on photosynthesis and the extent of protoplast volume reduction which occurs in leaves at low water potential (Psi(w)). This relationship was monitored in two cultivars (;Condor' and ;Capelle Desprez') of cultivated wheat (Triticum aestivum) that differed in sensitivity to drought, and in a wild relative of cultivated wheat (Triticum kotschyi) that has been previously found to be ;drought resistant.' When subjected to periods of water stress, Condor and T. kotschyi plants underwent osmotic adjustment; Capelle plants did not. Photosynthetic capacity was maintained to different extents in the three genotypes as leaf Psi(w) declined during stress; Capelle plants were most severely affected. Calculations of internal leaf [CO(2)] and stomatal conductance from gas exchange measurements indicated that differences in photosynthetic inhibition at low Psi(w) among the genotypes were primarily due to nonstomatal effects. The extent of protoplast volume reduction that occurred in leaves at low Psi(w) was also found to be different in the three genotypes; maintenance of protoplast volume and photosynthetic capacity in stressed plants of the genotypes appeared to be correlated. When the extent of water stress-induced inhibition of photosynthesis was plotted as a function of declining protoplast volume, this relationship appeared identical for the three genotypes. It was concluded that there is a correlative association between protoplast volume and photosynthetic capacity in leaves of wheat plants subjected to periods of water stress.

Journal Article↗
PubMed1989

Development and use of chlorotetracycline fluorescence as a measurement assay of chloroplast envelope-bound mg.

Experiments were conducted to develop chlorotetracycline (CTC) fluorescence as an assay of Mg(2+) bound to the envelope of the intact chloroplast. This assay technique has been widely used to measure envelope associated divalent cations in animal cell and subcellular systems, but has not been used with chloroplasts. Chloroplast envelope-associated Mg(2+) was altered by pretreatment with Mg(2+) and divalent cation chelating agents and by additions of Mg(2+) to the CTC assay medium. Results indicated that for a given chloroplast preparation, relative changes in envelope-associated Mg(2+) can be effectively monitored with CTC fluorescence. It was concluded that the limitations of this assay system are: (a) chlorophyll strongly quenches CTC fluorescence signal, so a constant chlorophyll concentration must be maintained, (b) measurements must be made quickly, and (c) use of the technique to compare different chloroplast preparations may not be valid. Studies with (28)Mg(2+) confirmed our interpretation of the fluorescence results, and also suggested that the chloroplast envelope is fairly impermeable to Mg(2+). It was concluded that changes in Mg(2+) associated with the chloroplast due to incubation of plastids in solutions containing up to 5 millimolar Mg(2+) may be exclusively due to increased envelope-associated Mg(2+). The CTC assay was used in experiments to demonstrate that increases in chloroplast envelope-associated Mg(2+) inhibit photosynthetic capacity. This inhibition can be partially overcome by the presence of K(+) in the photosynthetic reaction media.

Journal Article↗
PubMed1989

Maintenance of photosynthesis at low leaf water potential in wheat : role of potassium status and irrigation history.

The interaction of low water potential effects on photosynthesis, and leaf K(+) levels in wheat (Triticum aestivum L.) plants was studied. Plants were grown at three K(+) fertilization levels; 0.2, 2, and 6 millimolar. With well watered plants, 2 millimolar K(+) supported maximal photosynthetic rates; 0.2 millimolar K(+) was inhibitory, and 6 millimolar K(+) was superoptimal (i.e. rates were no greater than at 2 millimolar K(+)). Photosynthesis was monitored at high (930 parts per million) and low (330 parts per million) external CO(2) throughout a series of water stress cycles. Plants subjected to one stress cycle were considered nonacclimated; plants subjected to two successive cycles were considered acclimated during the second cycle. Sensitivity of photosynthesis to declining leaf water potential was affected by K(+) status; 6 millimolar K(+) plants were less sensitive, and 0.2 millimolar K(+) plants were more sensitive than 2 millimolar K(+) plants to declining water potential. This occurred with nonacclimated and acclimated plants at both high and low assay CO(2). It was concluded that the K(+) effect on photosynthesis under stress was not mediated by treatment effects on stomatal resistance. Differences between the K(+) treatments were much less pronounced, however, when photosynthesis of nonacclimated and acclimated plants was plotted at a function of declining relative water content during the stress cycles. These results suggest that K(+) effects on the relationship between relative water content and water potential in stressed plants was primarily responsible for the bulk of the K(+)-protective effect on photosynthesis in stressed plants. In vitro experiments with chloroplasts and protoplasts isolated from 2 millimolar K(+) and 6 millimolar K(+) plants indicated that upon dehydration, K(+) efflux from the chloroplast stroma into the cytoplasm is less pronounced in 6 millimolar K(+) protoplasts.

Journal Article↗
PubMed1989

Protoplast volume:water potential relationship and bound water fraction in spinach leaves.

Methods used to estimate the (nonosmotic) bound water fraction (BWF) (i.e. apoplast water) of spinach (Spinacia oleracea L.) leaves were evaluated. Studies using three different methods of pressure/volume (P/V) curve construction all resulted in a similar calculation of BWF; approximately 40%. The theoretically derived BWF, and the water potential (Psi(w))/relative water content relationship established from P/V curves were used to establish the relationship between protoplast (i.e. symplast) volume and Psi(w). Another method of establishing the protoplast volume/Psi(w) relationship in spinach leaves was compared with the results from P/V curve experiments. This second technique involved the vacuum infiltration of solutions at a range of osmotic potentials into discs cut from spinach leaves. These solutions contained radioactively labeled H(2)O and sorbitol. This dual label infiltration technique allowed for simultaneous measurement of the total and apoplast volumes in leaf tissue; the difference yielded the protoplast volume. The dual label infiltration experiments and the P/V curve constructions both showed that below -1 megapascals, protoplast volume decreases sharply with decreasing water potential; with 50% reduction in protoplast volume occurring at -1.8 megapascals leaf water potential.

Journal Article↗
PubMed1988

Antitranspirant associated abscisic Acid effects on the water relations and yield of transplanted bell peppers.

Greenhouse and field experiments were performed to determine if increased leaf resistance induced by exogenous application of abscisic acid (ABA) could enhance the water status of transplanted bell pepper seedlings. Seedling survival and yield were also monitored in the field experiment. When seedlings were transplanted into either wet or dry potting mix in the greenhouse, ABA increased leaf resistance and leaf water potential. In the field, plots were irrigated either immediately after, or 1 day after transplanting. Under both treatments, ABA application resulted in increased leaf resistance and water potential, but seedling survival and yield were enhanced due to ABA only in plots which were irrigated 1 day after transplanting. It is concluded that antitranspirant application can reduce transplant shock and increase yield of bell pepper.

Journal Article↗
PubMed1988

Chloroplast osmotic adjustment and water stress effects on photosynthesis.

Previous studies have suggested that chloroplast stromal volume reduction may mediate the inhibition of photosynthesis under water stress. In this study, the effects of spinach (Spinacia oleracea, var ;Winter Bloomsdale') plant water deficits on chloroplast photosynthetic capacity, solute concentrations in chloroplasts, and chloroplast volume were studied. In situ (gas exchange) and in vitro measurements indicated that chloroplast photosynthetic capacity was maintained during initial leaf water potential (Psi(w)) and relative water content (RWC) decline. During the latter part of the stress period, photosynthesis dropped precipitously. Chloroplast stromal volume apparently remained constant during the initial period of decline in RWC, but as leaf Psi(w) reached -1.2 megapascals, stromal volume began to decline. The apparent maintenance of stromal volume over the initial RWC decline during a stress cycle suggested that chloroplasts are capable of osmotic adjustment in response to leaf water deficits. This hypothesis was confirmed by measuring chloroplast solute levels, which increased during stress. The results of these experiments suggest that stromal volume reduction in situ may be associated with loss of photosynthetic capacity and that one mechanism of photosynthetic acclimation to low Psi(w) may involve stromal volume maintenance.

Journal Article↗
PubMed1987

Modulation of water stress effects on photosynthesis by altered leaf k.

Wheat irrigated with nutrient solutions containing 0, 0.2, 0.5, 1, 2, or 6 millimolar K(+) had maximum photosynthetic rates at 1 to 2 millimolar K(+) concentrations. Rates in the 6 millimolar K(+)-grown plants were not higher than the 2 millimolar K(+)-grown wheat, and rates were inhibited below 0.5 millimolar K(+). Photosynthesis was measured by both attached whole leaf CO(2) uptake and by (14)CO(2) fixation of leaf slices in solution. Exposure of leaf slices from 0.2, 2, and 6 millimolar K(+)-grown wheat to various assay media water potentials showed that photosynthesis of the 0.2 millimolar K(+)-grown wheat decreased from control (high water potential) rates by 35%, that of the 2 millimolar K(+)-grown wheat by 20.4%, and that of the 6 millimolar K(+)-grown wheat by only 8.3% at -3.11 megapascals. Also, photosynthesis of the 6 millimolar K(+)-grown wheat was enhanced by 28% over that of the 2 millimolar K(+) wheat at the most severe water stress (-3.11 megapascals), indicating that the excess leaf K(+) in the 6 millimolar K(+)-grown wheat partially reversed dehydration effects on photosynthesis. Oligomycin eliminated the protective effects of high K(+) on photosynthesis in dehydrated leaf slices. These results suggest that the protective effect of high K(+) under water stress may involve the exchange of K(+) in the cytoplasm for stroma H(+), thus altering stromal pH and restoring photosynthesis. The protective effect of high K(+) was also observed in attached whole leaf photosynthesis of in situ water-stressed wheat grown on 0.2, 2, and 6 millimolar K(+). Under water stress, rates of the 6 millimolar K(+)-grown wheat were enhanced by 66.2% and 113.9% over that of 2 millimolar K(+)-grown wheat in two separate experiments. Internal CO(2) concentration of the 6 millimolar K(+)-grown wheat was lower than that of the 0.2 and 2 millimolar K(+)-grown wheat. These results suggest that the high K(+) effects on chloroplast photosynthesis seen in leaf slices also occur at the whole plant level.

Journal Article↗
PubMed1987

Osmotic adjustment, symplast volume, and nonstomatally mediated water stress inhibition of photosynthesis in wheat.

At low water potential (psi(w)), dehydration reduces the symplast volume of leaf tissue. The effect of this reduction on photosynthetic capacity was investigated. The influence of osmotic adjustment on this relationship was also examined. To examine these relationships, comparative studies were undertaken on two wheat cultivars, one that osmotically adjusts in response to water deficits (;Condor'), and one that lacks this capacity (;Capelle Desprez'). During a 9-day stress cycle, when water was withheld from plants grown in a growth chamber, the relative water content of leaves declined by 30% in both cultivars. Leaf osmotic potential (psi(s)) declined to a greater degree in Condor plants. Measuring psi(s) at full turgor indicated that osmotic adjustment occurred in stressed Condor, but not in Capelle plants. Two methods were used to examine the degree of symplast (i.e. protoplast) volume reduction in tissue rapidly equilibrated to increasingly low psi(w). Both techniques gave similar results. With well-watered plants, symplast volume reduction from the maximum (found at high psi(w) for each cultivar) was the same for Condor and Capelle. After a stress cycle, volume was maintained to a greater degree at low psi(w) in Condor leaf tissue than in Capelle. Nonstomatally controlled photosynthesis was inhibited to the same degree at low psi(w) in leaf tissue prepared from well-watered Condor and Capelle plants. However, photosynthetic capacity was maintained to a greater degree at low psi(w) in tissue prepared from stressed Condor plants than in tissue from stressed Capelle plants. Net CO(2) uptake in attached leaves was monitored using an infrared gas analyzer. These studies indicated that in water stressed plants, photosynthesis was 106.5% higher in Condor than Capelle at ambient [CO(2)] and 21.8% higher at elevated external [CO(2)]. The results presented in this report were interpreted as consistent with the hypothesis that there is a causal association between protoplast (and presumably chloroplast) volume reduction at low psi(w) and low psi(w) inhibition of photosynthesis. Also, the data indicate that osmotic adjustment allows for maintenance of relatively greater volume at low psi(w), thus reducing low psi(w) inhibition of chloroplast photosynthetic potential.

Journal Article↗
PubMed1985

Leaf k interaction with water stress inhibition of nonstomatal-controlled photosynthesis.

The relationship between leaf K(+) concentration, in vitro dehydration, and nonstomatal-controlled photosynthesis was investigated using leaf slices that were vacuum infiltrated with media containing varying sorbitol concentrations. The leaf slices were from plants either supplied with complete or K(+)-deficient medium throughout a 35-day growth period. During this time, leaf K(+) concentration, water potential, osmotic potential, and turgor pressure were monitored. Leaf K(+) concentration averaged 239 micomoles per gram (fresh weight) in control plants, and dropped to 74.3 micromoles per gram (fresh weight) in K(+)-deficient plants. Less negative osmotic potentials and resultant turgor loss in K(+)-deficient plants indicated that the osmotically active pool of cellular K(+) was lower in those plants.The decrease in leaf K(+) concentration enhanced the dehydration inhibition of photosynthesis. For example, increasing sorbitol from 0.33 to 0.5 molar during incubation inhibited photosynthesis in the controls by 14% or less. This same protocol resulted in an inhibition of photosynthesis by as much as 41% in K(+)-deficient tissue. In contrast to the data obtained with leaf slices, dehydration inhibition of isolated chloroplast photosynthesis was not affected by K(+) status of parent plant material. These data are consistent with the hypothesis that one effect of leaf K(+) deficiencies on photosynthetic response to dehydration may be mediated by extra-choloroplastic factors.Ammonium ions, which facilitate stromal alkalinization, reversed the increased sensitivity of K(+)-deficient leaf slice photosynthesis to cell dehydration. However, NH(4) (+) had no effect on photosynthesis of K(+)-deficient leaf slices under nonhypertonic conditions. These data suggest that endogenous extra-chloroplastic K(+) may modulate dehydration inhibition of photosynthesis, possibly by facilitating stromal alkalinization.

Journal Article↗
PubMed1983

Reduced osmotic potential effects on photosynthesis : identification of stromal acidification as a mediating factor.

Addition of sorbitol, which facilitated reductions in reaction medium osmotic potential from standard (0.33 molar sorbitol, -10 bars) isotonic conditions to a stress level of 0.67 molar sorbitol (-20 bars), inhibited the photosynthetic capacity of isolated spinach (Spinacia oleracea) chloroplasts. This inhibition, which ranged from 64 to 74% under otherwise standard reaction conditions, was dependent on reaction medium inorganic phosphate concentration, with the phosphate optimum for photosynthesis reduced to 0.05 millimolar at the low osmotic potential stress treatment from a value of 0.25 millimolar under control conditions.Stromal alkalating agents such as NH(4)Cl (0.75 millimolar) and KCl (35 millimolar) were also found to affect the degree of low osmotic potential inhibition of photosynthesis. Both agents doubled the rate of NaHCO(3)-supported O(2) evolution under the stress treatment, while hardly affecting the control rate at optimal concentrations. These agents also reduced the length of the lag phase of photosynthetic O(2) evolution under the stress treatment to a much greater degree. The rate-enhancement effect of these agents under the stress treatment was reversed by sodium acetate, which is known to facilitate stromal acidification.The reaction medium pH optimum for photosynthesis under the stress treatment was higher than under control conditions. In the presence of optimal NH(4)Cl, this shift was no longer evident.Internal pH measurements indicated that the stress treatment caused a 0.43 and 0.24 unit reduction in the stromal and intrathylakoid pH, respectively, under illumination. This osmotically induced acidification was not evident in the dark. The presence of 0.75 millimolar NH(4)Cl partially reversed the osmotically induced reduction in the illuminated stromal pH. It was concluded that stromal acidification is a mediating mechanism of the most severe site of low osmotic potential inhibition of the photosynthetic process.

Journal Article↗
PubMed1983

Reduced osmotic potential inhibition of photosynthesis : site-specific effects of osmotically induced stromal acidification.

The effects of reduced reaction medium osmotic potential (0.67 molar sorbitol as compared to a control treatment with 0.33 molar sorbitol) on the enzymic steps of the photosynthetic carbon reduction cycle were investigated using isolated spinach (Spinacia oleracea L. var Longstanding Bloomsdale) chloroplasts. Reversal of reduced osmotic potential inhibition of photosynthetic rates by a stromal alkalating agent (NH(4)Cl) was associated with specific steps of the cycle. Low osmotic potential induced stromal acidification was found to be facilitated by osmotically induced chloroplast shrinkage. However, the action of the alkalating agent was found not to be associated with reversal of osmotically induced morphological changes of the stromal compartment.Labeled metabolite analyses indicated that the osmotic stress treatment caused the substrate for fructose 1,6-bisphosphatase (FBPase) to build up in the absence of NH(4)Cl, and the substrate for phosphoribulokinase to increase in the presence of NH(4)Cl. These data were interpreted as indicating that the most severe effect of osmotic stress on photosynthesis is at the site of FBPase, and that this inhibition is mediated by osmotically induced stromal acidification. Phosphoribulokinase activity inhibition at the low osmotic potential treatment was apparently less severe and not mediated by stromal acidification. A third site of osmotic inhibition, which was reversed by NH(4)Cl, and therefore was assumed to be mediated by stromal acidification, was at the step of ribulose 1,5-bisphosphate carboxylase.Additions of NH(4)Cl also enhanced the activity of the pH-insensitive phase of the photosynthetic carbon reduction cycle, 3-phosphoglyceric acid reduction, at the stress treatment. This effect was thought to be mediated by the removal of the block at FBPase. A model was proposed to outline the relative severity of osmotic stress effects at various sites of the photosynthetic carbon reduction cycle.

Journal Article↗
PubMed1983

Stromal acidification mediates in vivo water stress inhibition of nonstomatal-controlled photosynthesis.

Stromal acidification has been reported to mediate reduced osmotic potential (psi(pi)) effects on photosynthesis in the isolated spinach chloroplast (Berkowitz, Gibbs 1983 Plant Physiol 72: 1100-1109). To determine if stromal acidification mediates osmotic dehydration inhibition of photosynthesis in vivo, the effects of a weak base (NH(4)Cl), which raises stromal pH, on CO(2) fixation of vacuum-infiltrated spinach leaf slices, Chlamydomonas reinhardii cells and Aphanocapsa 6308 cells under isotonic and dehydrating conditions were investigated. Five millimolar NH(4)Cl stimulated spinach leaf slice CO(2) fixation by 43% under stress (0.67 molar sorbitol) conditions, and had little effect on fixation under isotonic (0.33 molar sorbitol) conditions. Chlamydomonas cells were found to be more sensitive to reduced psi(pi) than spinach leaf slices. CO(2) fixation in the cells of the green alga Chlamydomonas reinhardii was 99 and 17 micromoles per milligram chlorophyll per hour, respectively, at 0.1 molar mannitol and 0.28 molar mannitol. Five millimolar NH(4)Cl stimulated CO(2) fixation of Chlamydomonas cells by 147% under stress (0.28 molar mannitol) conditions. Aphanocapsa 6308 cells (blue-green alga) were also found to be sensitive to reduced psi(pi), and inhibitions in photosynthesis were partially reversed by NH(4)Cl. These data indicate that in vivo water stress inhibition of photosynthesis is facilitated by stromal acidification, and that this inhibition can be at least partially reversed in situ.

Journal Article↗
PubMed1982

Effect of osmotic stress on photosynthesis studied with the isolated spinach chloroplast : generation and use of reducing power.

The effect of increasing assay medium sorbitol concentration from 0.33 to 1.0 molar on the photosynthetic reactions of intact and broken spinach (Spinacia oleracea L. var. Long Standing Bloomsdale) chloroplasts was investigated by monitoring O(2) evolution supported by the addition of glyceric acid 3-phosphate (PGA), oxaloacetic acid (OAA), 2,5-dimethyl-p-benzoquinone, and 2,6-dichlorophenolindophenol or as O(2) uptake with methyl viologen as acceptor.Uncoupled 2,6-dichlorophenolindophenol-supported whole chain electron transport (photosystems I and II) was inhibited from the 0.33 molar rate by 14% and 48.6% at 0.67 and 1.0 molar sorbitol in the intact chloroplast and by only 0.4% and 25.0% in the broken chloroplast preparation. Whole chain electron flow from water to other oxidants (OAA, methyl viologen) was also inhibited at increased osmoticum in intact preparations while electron flow from water to methyl viologen, ferricyanide, and NADP in broken preparations did not demonstrate the osmotic response. Electron transport to 2,5-dimethyl-p-benzoquinone (photosystem II) from H(2)O and to methyl viologen (photosystem I) from 3,3'-diaminobenzidine were found to be unaffected by osmolarity in both intact and broken preparations.The stress response was more pronounced (26-38%) with PGA as substrate in the presence of 0.67 molar sorbitol than the inhibition found with uncoupled and coupled linear electron flow. In addition, substrate availability and ATP generated by cyclic photophosphorylation evaluated by addition of Antimycin A were found not to be mediating the full osmotic inhibition of PGA-supported O(2) evolution. In a reconstituted (thylakoids plus stromal protein) chloroplast system to which a substrate level of PGA was added, O(2) evolution was only slightly (7.8%) inhibited by increased osmolarity (0.33-0.67 molar sorbitol) indicating that the level of osmotic inhibition above that contributed by adverse effects on electron flow can be attributed to the functioning of the photosynthetic carbon reduction cycle within the intact chloroplasts.

Journal Article↗
PubMed1982

Effect of osmotic stress on photosynthesis studied with the isolated spinach chloroplast : site-specific inhibition of the photosynthetic carbon reduction cycle.

The effects of reduced osmotic potential on the photosynthetic carbon reduction cycle were investigated by monitoring photosynthetic processes of spinach (Spinacia oleracea L. var. Long Standing Bloomsdale) chloroplasts exposed to increased assay medium sorbitol concentrations. CO(2) assimilation was found to be inhibited at 0.67 molar sorbitol by about 60% from control rates at 0.33 molar sorbitol. This level of stress inhibition was greater than that affecting the reductive phase of the cycle; glycerate 3-phosphate reduction was inhibited at 0.67 molar by 27 to 40%. Sorbitol (0.67 molar) inhibited the rate of O(2) evolution at saturating and limiting concentrations of NaHCO(3), and extended the lag phase of O(2) evolution. This indicated that factors which are rate-limiting to the photosynthetic process are adversely affected by reduced osmotic potential.Analysis of photosynthetic products following CO(2) fixation in 0.33 molar sorbitol and 0.67 molar sorbitol indicated that reduced osmotic potential facilitated increases in the levels of fructose 1,6-bisphosphate and triose phosphates with reductions in glucose 6-phosphate and fructose 6-phosphate, implicating fructose 1,6-bisphosphatase as a site of osmotic stress. Osmotic inhibition of the reductive portion (glycerate 3-phosphate to triose phosphate) of the photosynthetic carbon reduction cycle was partially attributed to feedback inhibition by the product, triose phosphate, on glycerate 3-phosphate reduction. A saturating concentration of ribose 5-phosphate partially overcame osmotic inhibition of CO(2)-supported O(2) evolution, indicating another but apparently less severe site of stress inhibition in the sequence of ribose 5-phosphate to glycerate 3-phosphate.

Journal Article↗