The Path of Carbon Flow during NO(3)-Induced Photosynthetic Suppression in N-Limited Selenastrum minutum

Nitrate addition to nitrate-limited cultures of Selenastrum minutum Naeg. Collins (Chlorophyta) resulted in a 70% suppression of photosynthetic carbon fixation. In ¹⁴CO₂ pulse/chase experiments nitrate resupply increased radiolabel incorporation into amino and organic acids and decreased radiolabel incorporation into insoluble material. Nitrate resupply increased the concentration of phosphoenolpyruvate and increased the radiolabeling of phosphoenolpyruvate, pyruvate and tricarboxylic acid cycle intermediates, notably citrate, fumarate, and malate. Furthermore, nitrate also increased the pool sizes and radiolabeling of most amino acids, with alanine, aspartate, glutamate, and glutamine showing the largest changes. Nitrate resupply increased the proportion of radiolabel in the C-4 position of malate and increased the ratios of radiolabel in aspartate to phosphoenolpyruvate and in pyruvate to phosphoenolpyruvate, indicative of increased phosphoenolpyruvate carboxylase and pyruvate kinase activities. Analysis of these data showed that the rate of carbon flow through glutamate (10.6 μmoles glutamate per milligram chlorophyll per hour) and the rate of net glutamate production (7.9 μmoles glutamate per milligram chlorophyll per hour) were both greater than the maximum rate of carbon export from the Calvin cycle which could be maintained during steady state photosynthesis. These results are consistent with the hypothesis that nitrogen resupply to nitrogen-limited microalgae results in a transient suppression of photosynthetic carbon fixation due, in part, to the severity of competition for carbon skeletons between the Calvin cycle and nitrogen assimilation (IR Elrifi, DH Turpin 1986 Plant Physiol 81: 273-279).     

Gradients of Intercellular CO(2) Levels Across the Leaf Mesophyll

Most current photosynthesis models, and interpretations of many wholeleaf CO₂ gas exchange measurements, are based on the often unstated assumption that the partial pressure of CO₂ is nearly uniform throughout the airspaces of the leaf mesophyll. Here we present measurements of CO₂ gradients across amphistomatous leaves allowed to assimilate CO₂ through only one surface, thus simulating hypostomatous leaves. We studied five species: Eucalyptus pauciflora Sieb. ex Spreng., Brassica chinensis L., Gossypium hirsutum L., Phaseolus vulgaris L., and Spinacia oleracea L. For Eucalyptus, maximum CO₂ pressure differences across the leaf mesophyll were 73 and 160 microbar when the pressures outside the lower leaf surface were 310 and 590 microbar, respectively. Using an approximate theoretical calculation, we infer that if the CO₂ had been supplied equally at both surfaces then the respective mean intercellular CO₂ pressures would have been roughly 12 and 27 microbar less than the pressures in the substomatal cavities in these cases. For ambient CO₂ pressures near 320 microbar, the average and minimum pressure differences across the mesophyll were 45 and 13 microbar. The corresponding mean intercellular CO₂ pressures would then be roughly 8 and 2 microbar less than those in the substomatal cavities. Pressure differences were generally smaller for the four agricultural species than for Eucalyptus, but they were nevertheless larger than previously reported values.     

Further evidence of oxygen diffusion as the determining factor in the relation between disk thickness and respiration of potato tissue

The effect of oxygen tension above atmospheric pO₂ on the development of respiratory capacity in potato disks has been examined. Raising the oxygen tension of the aqueous environment to 40% during the aging of 2.0 mm or 3.0 mm thick disks at 25° progressively increased the respiration rate of the tissue as shown by subsequent assay in 100% oxygen. Disks 3.0 mm thick showed a greater response to increased pO₂ than did 2.0 mm disks. A comparison of center 1.0 mm sections excised from 3.0 mm disks after aging, showed that the respiration rate of internal tissue from disks aged in high pO₂ was approximately 40% greater than such tissue aged with atmospheric pO₂. The characteristic inverse relationship between respiration rate and thickness in aged disks can be modified from a concave-downwards curve to a convex-downwards curve by pretreating the tissue with increased pO₂, thus indicating that raising the pO₂ during aging can increase the thickness threshold at which the transition from tissue manifesting the respiratory characteristics of thin disks to that manifesting the characteristics of thick disks, occurs. Similarly increased pO₂ during aging can modify the hyperbolic relationship obtaining between pretreatment temperature in the range 10° to 25° and respiratory capacity of aged 3.0 mm disks, to approximate to the linear relationship observed with 0.75 mm disks. It is concluded that the development of respiratory capacity in disks between 0.75 mm and 3.0 mm thick is restricted by oxygen dificiency and that the characteristic inverse relationship between respiration rate and thickness in aged disks is largely attributable to this factor, the influence of which is discernible both on the development of respiratory capacity and on its subsequent assay.     

A pre-formed Pyrogenic Factor Released by Lipopolysaccharide Stimulated Macrophages

The aim of this study was to investigate the pyrogenic activity of factor(s) released by rat peritoneal macrophages following a brief stimulation with LPS. The effect of this factor on the number of circulating leukocytes and serum Fe, Cu and Zn levels, was also evaluated. The possibility that the content of interleukin (IL)-1beta, IL-6 and tumour necrosis factor (TNF) in the supernatant could explain the observations was investigated. Supernatant produced over a period of 1 h by peritoneal macrophages, following a 30 min incubation with LPS at 37 degrees C, was ultrafiltered through a 10 000 MW cut-off Amicon membrane, sterilized, and concentrated 2.5, 5, 10 and 20 times. The intravenous (i.v.) injection of this supernatant induced a concentration-dependent fever in rats with a maximal response at 2 h. The pyrogenic activity was produced by macrophages elicited with thioglycollate and by resident cells. The supernatants also induced neutrophilia and reduction in Fe and Zn 6 h after the injection. Absence of activity in boiled supernatants, or supernatants from macrophages incubated at 4 degrees C with LPS, indicates that LPS was not responsible for the activity. In vitro treatment with indomethacin (Indo), dexamethasone (Dex), or cycloheximide (Chx) did not modify the release of pyrogenic activity into the supernatant or its effects on the reduction in serum metal levels. Although Chx abolished the production of mediator(s) inducing neutrophilia, and Dex reduced the induction of IL-1beta, TNF and IL-6, injection of the highest concentration of these cytokines detected in the supernatants did not induce fever. In vivo treatment with Dex, but not Indo, abolished the fever induced by the supernatant. These results suggest that macrophages contain pre-formed pyrogenic mediator(s), not related to IL-1beta, IL-6 or TNF, that acts indirectly and independently of prostaglandtn. It also seems likely that the pyrogenic activity is related to the factor responsible for the reduction of serum Fe and Zn levels, but not the neutrophilia.     

The protein coats or ghosts or coliphage T2. III. Metabolic studies of Escherichia coli B infected with T2 bacteriophage ghosts

Assimilation of oxygen, inorganic phosphate, and ammonia nitrogen by normal T2 phage and T2 ghost-infected E. coli B was studied. The rate of oxygen and phosphorus uptake by ghost-infected bacteria is similar to that of normal and phage-infected cells. The R.Q. in glucose-salts medium remains approximately 1. Assimilation of ammonia nitrogen by ghost-infected bacteria is maintained at a rate approximately 80 per cent of normal. The inorganic phosphate which is assimilated was found to be incorporated into TCA-soluble compounds which were rapidly released into the medium. Within 5 minutes after absorption of the ghosts there was a loss from the cell of TCA-soluble constituents including organic phosphorus and compounds which absorb at 260 mmicro. No corresponding breakdown of nucleic acid present in the cell prior to infection could be detected. The incorporation of inorganic phosphate into organic linkages in the ghost-infected cell and its release into the medium were found to proceed at a rate approaching that of the incorporation of inorganic phosphorus into the nucleic acid of normal cells. The net increase in 260 mmicro absorbing compounds appeared to be inhibited.     

Effects of Iron Limitation on Photosystem II Composition and Light Utilization in Dunaliella tertiolecta

The effects of iron limitation on photosystem II (PSII) composition and photochemical energy conversion efficiency were studied in the unicellular chlorophyte alga Dunaliella tertiolecta. The quantum yield of photochemistry in PSII, inferred from changes in variable fluorescence normalized to the maximum fluorescence yield, was markedly lower in iron-limited cells and increased 3-fold within 20 h following the addition of iron. The decrease in the quantum yield of photochemistry was correlated with increased fluorescence emission from the antenna. In iron-limited cells, flash intensity saturation profiles of variable fluorescence closely followed a cumulative one-hit Poisson model, suggesting that PSII reaction centers are energetically isolated, whereas in iron-replete cells, the slope of the profile was steeper and the calculated probability of energy transfer between reaction centers increased to >0.6. Immunoassays revealed that in iron-limited cells the reaction center proteins, D1, CP43, and CP47, were markedly reduced relative to the peripheral light-harvesting Chl-protein complex of PSII, whereas the [alpha] subunit of cytochrome b559 was about 10-fold higher. Spectroscopic analysis established that the cytochrome b559 peptide did not contain an associated functional heme. We conclude that the photochemical conversion of absorbed excitation energy in iron-limited cells is limited by the number of photochemical traps per unit antenna.     

Is There a Role for the Apex in Shoot Geotropism?

Experiments with horizontal etiolated sunflower (Helianthus annuus L.) seedlings supported centrally such that both apical and basal ends are free to react to geostimulus, revealed that the apical end commences curvature 1 to 2 hours earlier than the basal end. The later curvature in the basal region is a consequence of the absence of growth in the initial period rather than merely slower growth. A comparison of zonal growth rates in a vertical and a horizontal seedling confirmed that geostimulus induces a renewal of growth in a region where growth had ceased. Removing the apical half of the hypocotyl showed that the curvature resulting from this growth initiation in the basal region is dependent on attachment to the apical region. Evidence that this dependence is unlikely to be due to energy deficiency is adduced. The prior response of the apical end to geostimulus and the apically dependent later initiation of new growth in the basal region are compatible with the delay inherent in message transport from apex to base and are considered as evidence for apical involvement in the totality of the seedling's georesponse.