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  • Life and Medical Sciences  (85)
  • Autoradiography  (6)
  • Photorespiration  (6)
  • Pisum  (6)
  • 1
    ISSN: 1432-2048
    Keywords: Glutamine synthetase ; Glutamate synthase ; Hordeum (photorespiration) ; Photorespiration ; Photorespiratory mutants (barley) ; Photosynthesis (electron transport, CO2 assimilation)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Heterozygous plants of barley (Hordeum vulgare L. cv. Maris Mink) with activities of chloroplastic glutamine synthetase (GS) between 47‰ and 97‰ of the wild-type and ferredoxin-dependent glutamate synthase (Fd-GOGAT) activities down to 63‰ of the wild-type have been used to study the control of photosynthetic fluxes. Rates of CO2 assimilation measured over a range of intercellular CO2 concentrations and photon flux densities (PFDs) were little different in the wild-type and a mutant with 47‰ GS, although total activities of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) decreased by about 20‰ with a decrease in GS to 50‰ of the wild-type. The quantum efficiencies of photosystem II electron transport (ΦPSII and CO2 assimilation ΦCO2) were determined. ΦPSII was lower than expected in mutants with 50‰ less GS under conditions which enhance the photorespiratory flux, but were identical to the wild-type under non-photorespiratory conditions, suggesting that at high rates of photorespiration the electron requirement for net CO2 assimilation declines in plants with decreased GS. This discrepancy in the electron requirement between the wild-type and the 47‰ GS mutant was enhanced at high temperatures and low CO2, conditions which favour oxygenation by Rubisco. Photochemical and non-photochemical chlorophyll a fluorescence quenching as well as the quantum efficiency of excitation-energy capture by open photosystem II reaction centres were differentially affected in mutants with less GS relative to the wild-type when CO2 was lowered or the PFD was varied. The quantum efficiencies of electron transport in photosystems I and II were closely correlated under a range of PFDs and CO2 concentrations, confirming that the rate of linear electron transport was much lower in plants with less GS. It is shown that GS exerts considerable control (flux control coefficients between 0.5 and 1.0) on the electron requirement for CO2 assimilation at high fluxes of photorespiration relative to CO2 assimilation. Apart from the control of GS on protein and Rubisco contents, GS in the wild-type has also some direct positive control on CO2 assimilation. However, negative control on CO2 assimilation was found in mutants with 50‰ less GS. These data, taken with the data on electron requirements for CO2 assimilation, suggest that CO2-fixing processes other than that catalysed by Rubisco, such as carboxylation of phosphoenolpyruvate, or an inhibition of photorespiration (e.g. glycine decarboxylation), may contribute to the observed CO2 exchange and photosystem II electron transport in plants with less GS. In the 63‰-Fd-GOGAT mutant, rates of CO2 assimilation were appreciably lower than in the wild-type under a range of PFDs and CO2 concentrations, which largely reflected lower contents of Rubisco in the Fd-GOGAT mutants. Assimilation of CO2 was inhibited appreciably at high CO2 concentrations. There was little difference in the electron requirement for CO2 assimilation between the wild-type and mutants with less Fd-GOGAT, although there were indications that a triose-phosphate/glycerate-3-phosphate shuttle or cyclic electron transport operates to balance ATP generation and NADP reduction. The latter was supported by a curvilinear relationship of photosystem I and II electron transport in the 63‰ Fd-GOGAT mutant. A positive control is exerted by Fd-GOGAT on the amounts of protein and Rubisco and on CO2 assimilation.
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  • 2
    ISSN: 1432-2048
    Keywords: Key words: Hordeum (mutants) ; Glycine decarboxylase ; Glyoxylate ; Mutant (barley) ; Photosynthesis ; Photorespiration
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract. A mutant (LaPr 87/30) of barley (Hordeum vulgare L.) deficient in glycine decarboxylase (GDC; EC 2.1.2.10) was crossed with wild-type plants to generate heterozygous plants with reduced GDC activities. Plants of the F2 generation were grown in air and analysed for reductions in GDC proteins and GDC activity. The leaves of heterozygous plants contained reduced amounts of H-protein, and when the content of H-protein was lower than 60% of the wild-type, the P-protein was also reduced. The contents of the other two proteins of the GDC complex, T-protein and L-protein were not affected. Glycine decarboxylase activities, measured as the decarboxylation of [1-14C]glycine by intact mitochondria released from protoplasts, were between 47% and 63% of the wild-type activity in heterozygous plants and between 86% and 100% in plants with normal contents of H-protein. The enzyme activity was linearly correlated with the relative content of H-protein. Plants with reduced GDC activities developed normally and did not show major pleiotropic effects. In air, the reduction in GDC activity had no effect on the leaf metabolite content or photosynthesis, but under conditions of enhanced photorespiration (low CO2 and high light), glycine accumulated and the rates of photosynthesis decreased compared to the wild-type. The accumulation of glycine did not lead to a depletion of amino donors or to the accumulation of glyoxylate. The lower rates of photosynthesis were probably caused by an impaired recycling of carbon in the photorespiratory pathway. It is concluded that GDC has no control over CO2 assimilation under normal growth conditions, but appreciable control by GDC becomes apparent under conditions leading to higher rates of photorespiration.
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  • 3
    ISSN: 1432-2048
    Keywords: Apical senescence ; Gibberellins ; Photomorphogenesis ; Pisum ; Senescence
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract In an early-flowering line of pea (G2) apical senescence occurs only in long days (LD), while growth in short days (SD) is indeterminate. In SD, G2 plants are known to produce a graft-transmissible substance which delays apical senescence in related lines that are photoperiod-insensitive with regard to apical senescence. Gibberellic acid (GA3) applied to the apical bud of G2 plants in LD delayed apical senescence indefinitely, while N6-benzyladenine and α-naphthaleneacetic acid were ineffective. Of the gibberellins native to pea, GA9 had no effect whereas GA20 had a moderate senescence-delaying effect. [3H]GA9 metabolism in intact leaves of G2 plants was inhibited by LD and was restored by placing the plants back in SD. Leaves of photoperiod-insensitive lines (I-types) metabolized GA9 readily regardless of photoperiod, but the metabolites differed qualitatively from those in G2 leaves. A polar GA9 metabolite, GAE, was found only in G2 plants in SD. The level of GA-like substances in methanol extracts from G2 plants dropped about 10-fold after the plants were moved from SD to LD; it was restored by transferring the plants back to SD. A polar zone of these GA-like materials co-chromatographed with GAE. It is suggested that a polar gibberellin is synthesized by G2 plants in SD; this gibberellin promotes shoot growth and meristematic activity in the shoot apex, preventing senescence.
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  • 4
    Electronic Resource
    Electronic Resource
    Springer
    Planta 142 (1978), S. 211-219 
    ISSN: 1432-2048
    Keywords: Auxin transport ; Auxin uptake ; Pisum
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract 1. The uptake of indol-3-yl acetic acid ([1-14C]IAA, 0–2.0 μM) into light-grown pea stem segments was measured under various conditions to investigate the extent to which mechanisms of auxin transport in crown gall suspension culture cells (Rubery and Sheldrake, Planta 118, 101–121, 1974) are also found in a tissue capable of polar auxin transport. — 2. IAA uptake increased as the external pH was lowered. IAA uptake was less than that of benzoic acid (BA), naphthylacetic acid (NAA) or 2,4 dichlorophenoxyacetic acid (2,4D) under equivalent conditions. TIBA enhanced net IAA uptake through inhibition of efflux, and to a lesser extent, also increased uptake of NAA and 2,4D while it had no effect on BA uptake. — 3. Both DNP and, at higher concentrations, BA, reduced IAA uptake probably because of a reduction of cytoplasmic pH. However, low concentrations of both BA and DNP caused a slight enhancement of IAA net uptake, possibly through a reduction of carrier-mediated IAA efflux. In the presence of TIBA, the inhibitory effects of DNP and BA were more severe and there was no enhancement of uptake at low concentrations. — 4. Non-radioactive IAA (10 μM) reduced uptake of labelled IAA but further increases in concentration up to 1.0 mM produced first an inhibition (0–10 min) of labelled IAA uptake, followed by a stimulation at later times. Non-radioactive 2,4 D decreased, but was not observed to stimulate, uptake of labelled IAA. In the presence of TIBA labelled IAA uptake was inhibited by non-radioactive IAA regardless of its concentration. — 5. Sulphydryl reagents PCMB and PCMBS promoted or inhibited IAA uptake depending, respectively, on whether they penetrated or were excluded from the cells. The penetrant PCMB also reduced the promotion of labelled IAA uptake by TIBA or by high concentrations of added non-labelled IAA. — 6. Our findings are interpreted as being consistent with the diffusive entry of unionised IAA into cells together with some carrier-mediated uptake. Auxin efflux from the cells also appears to have a carrier-mediated contribution, at least part of which is inhibited by TIBA, and which has a capacity at least as great as that of the uptake carrier. The data indicate that pea stem segments contain cells whose mechanisms of trans-membrane auxin transport fit the model of polar auxin transport proposed from experiments with crown gall suspension cells, although differences, particularly of carrier specificity, are apparent between the two systems.
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  • 5
    ISSN: 1432-2048
    Keywords: Hordeum ; Glutamine synthetase ; Glyoxylate ; Glyoxylate metabolism ; Photorespiration ; Ribulose-1,5-bisphosphate carboxylase-oxygenase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Heterozygous mutants of barley (Hordeum vulgare L. cv. Maris Mink) with decreased activities of chloroplastic glutamine synthetase (GS) between 97 and 47% of the wild type and ferredoxin dependent glutamate synthase (Fd-GOGAT) down to 64% of the wild type have been used to study aspects of glyoxylate metabolism and the effect of glyoxylate on the activation state of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) in vivo. In the leaf, the extractable activities of serine:glyoxylate aminotransferase decreased with a decrease in GS whereas activities of glutamate and alanine:glyoxylate aminotransferase increased, pointing to a re direction of amino donors from serine to glutamate and alanine. Under ambient conditions, the leaf contents of glutamate and alanine declined continuously with a decrease in GS, in parallel with the decrease in total amino acids. Glycine, serine and asparagine contents decreased with a decrease in GS to approximately 70% of the wild type, but increased again with a further decrease in GS. At high irradiances and at low CO2 concentrations, glyoxylate contents exhibited a pronounced minimum between 60% and 80% GS. With a further decrease in GS, glyoxylate contents recovered and approached values similar to the wild type. The activation state of Rubisco showed a negative correlation with glyoxylate contents, indicating that a decrease in GS feeds back on the first step of carbon assimilation and photorespiration. The activation state of stromal fructose-1,6-bisphosphatase was unaffected by a decrease in GS or Fd-GOGAT, whereas the activation state of NADP dependent malate dehydrogenase changed in a complex manner. The CO2photocompensation point, Γ*, was appreciably increased in mutants with 47% GS. ‘Mitochondrial respiration’ in the light (Rd) was reduced with a decrease in GS. Relative rates of CO2 release into CO2-free air between the wild type and the 47%-GS mutant correlated with determinations of Γ*. These data are consistent with the view that when GS is decreased there is an increased oxidative decarboxylation of glyoxylate resulting from a decreased availability of amino donors for the transamination of glyoxylate to glycine, and that when GS activities are lower than 70% of the wild type an additional mechanism operates to reduce the photorespiratory loss of ammonia.
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  • 6
    ISSN: 1432-2048
    Keywords: Catalase deficiency ; Hordeum (mutant) ; Mutant (barley) ; Photorespiration
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A mutant line of barley, R(othamsted)-Pr 79/4, has been isolated which grows poorly in natural air, but normally in air enriched to 0.2% CO2. Analysis of the products of 14CO2 fixation showed that there was no major block in photosynthetic or photorespiratory carbon metabolism in the mutant and that rates of CO2 fixation were only slightly lower than those measured in the wild type (c.v. Maris Mink). Leaves of the mutant line contained only 10% of the catalase (EC 1.11.1.6) activity found in the wild type; and the two major bands of catalase activity detected after starch-gel electrophoresis of extracts of normal leaves were missing from similar extracts of RPr 79/4. Peroxisomes isolated from mutant leaves contained negligible catalase activity, but normal levels of other enzymes involved in photorespiration. Genetic analysis has shown that the mutation is recessive and that both air-sensitivity and catalase-deficiency segregate together in F2 plants derived from a cross between the mutant and the cultivar Golden Promise. [1-14C]Glycollate was not converted to 14CO2 faster in the mutant leaves than in the normal leaves. Thus there was no evidence that photorespiratory CO2 may be obtained by the chemical action of H2O2 on glyoxylate or hydroxypyruvate.
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  • 7
    ISSN: 1432-2048
    Keywords: Ferredoxin ; Glutamate synthase ; Greening (leaves) ; Hordeum ; Pisum
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The activity of NAD(P)H-dependent glutamate synthase (E.C. 1.4.1.14) has been demonstrated in extracts from etiolated shoots of pea (Pisum sativum L.) and barley (Hordeum vulgare L.). This activity does not significantly alter upon greening of the etiolated shoots, and is at a similar level in light-grown material. Ferredoxin-dependent glutamate synthase (E.C. 1.4.7.1) has low activity in etiolated shoots but increases rapidly on greening. In light grown leaves ferredoxin-dependent activity is 30–40-fold higher than NAD(P)H-dependent activity. It is not considered that the NAD(P)H-dependent glutamate synthase plays an important role in ammonia assimilation in the photosynthetic tissue of higher plants.
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  • 8
    ISSN: 1432-2048
    Keywords: Amino acid metabolism ; Glutamine synthetase ; Glutamate synthase ; Hordeum (photorespiration) ; Photorespiration ; Photorespiratory mutants (barley)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Wild-type and mutant plants of barley (Hordeum vulgare L. cv. Maris Mink) lacking activities of chloroplastic glutamine synthetase (GS) and of ferredox-in-dependent glutamate synthase (Fd-GOGAT) were crossed to generate heterozygous plants. Crosses of the F2 generation containing GS activities between 47‰ and 97‰ of the wild-type and Fd-GOGAT activities down to 63‰ of the wild-type have been selected to study the control of both enzymes on photorespiratory carbon and nitrogen metabolism. There were no major pleiotropic effects. Decreased GS had a small impact on leaf protein and the total activity of ribulose-1,5-bisphosphate carboxylase-oxygenase (Rubisco). The activation state of Rubisco was unaffected in air, but a decrease in GS influenced the activation state of Rubisco in low CO2. In illuminated leaves, the amino-acid content decreased with decreasing GS, while the content of ammonium rose, showing that even small reductions in GS limit ammonium re-assimilation and may bring about a loss of nitrogen from the plants, and hence a reduction in protein and Rubisco. Leaf amino-acid contents were restored, and ammonium and nitrate contents decreased, by leaving plants in the dark for 24 h. The ratios of serine to glycine decreased with a decrease in GS when plants were kept at moderate photon flux densities in air, suggesting a possible feedback on glycine decarboxylation. This effect was absent in high light and low CO2. Under these conditions ammonium contents exhibited an optimum and amino-acid contents a minimum at a GS activity of 65‰ of the wild-type, suggesting an inhibition of ammonium release in mutants with less than 65‰ GS. The leaf contents of glutamate, glutamine, aspartate, asparagine, and alanine largely followed changes in the total amino-acid contents determined under different environmental conditions. Decreased Fd-GOGAT resulted in a decrease in leaf protein, chlorophyll, Rubisco and nitrate contents. Chlorophyll a/b ratios and specific leaf fresh weight were lower than in the wild-type. Leaf ammonium contents were similar to the wild-type and total leaf amino-acid contents were only affected in low CO2 at high photon flux densities, but mutants with decreased Fd-GOGAT accumulated glutamine and contained less glutamate.
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  • 9
    ISSN: 1432-2048
    Keywords: Key words:Amaranthus (mutant) ; C1-tetrahydrofolate synthase pathway ; 10-Formyltetrahydrofolate synthetase ; Glycine decarboxylase ; Hordeum (mutant) ; Photorespiration
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract. Glycine-accumulating mutants of barley (Hordeum vulgare L.) and Amaranthus edulis (Speg.), which lack the ability to decarboxylate glycine by glycine decarboxylase (GDC; EC 2.1.2.10), were used to study the significance of an alternative photorespiratory pathway of serine formation. In the normal photorespiratory pathway, 5,10-methylenetetrahydrofolate is formed in the reaction catalysed by GDC and transferred to serine by serine hydroxymethyltransferase. In an alternative pathway, glyoxylate could be decarboxylated to formate and formate could be converted into 5,10-methylenetetrahydrofolate in the C1-tetrahydrofolate synthase pathway. In contrast to wild-type plants, the mutants showed a light-dependent accumulation of glyoxylate and formate, which was suppressed by elevated (0.7%) CO2 concentrations. After growth in air, the activity and amount of 10-formyltetrahydrofolate synthetase (FTHF synthetase; EC 6.3.4.4), the first enzyme of the conversion of formate into 5,10-methylenetetrahydrofolate, were increased in the mutants compared to the wild types. A similar increase in FTHF synthetase could be induced by incubating leaves of wild-type plants with glycine under illumination, but not in the dark. Experiments with 14C showed that the barley mutants incorporated [14C]formate and [2-14C]glycollate into serine. Together, the accumulation of glyoxylate and formate under photorespiratory conditions, the increase in FTHF synthetase and the ability to utilise formate and glycollate for the formation of serine indicate that the mutants are able partially to compensate for the lack of GDC activity by bypassing the normal photorespiratory pathway.
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  • 10
    ISSN: 1432-0878
    Keywords: LH-cells ; Prolactin cells ; Immunocytochemistry ; Estrogen ; Autoradiography ; Guinea pig ; Hamster ; Gerbil
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Nuclear uptake and retention of3H-estradiol by luteinizing hormone (LH) and prolactin (PRL) cells was examined in three species of rodents (guinea pigs, hamsters and gerbils) using the combined techniques of immunocyto-chemistry and autoradiography. Castrated animals were injected with3H-estradiol and decapitated 1.5 h later. The pituitary glands were processed for thaw-mount autoradiography followed by conventional immunocytochemical staining for LH and PRL.3H-estradiol accumulated in more than 80% of the anterior pituitary cells in the gerbils, while only 33 and 22% of the cells accumulated3H-estradiol in the hamsters and guinea pigs, respectively. A varying percentage of immunoreactive LH and PRL cells in all three species were found also to contain binding sites for estradiol. Some LH and PRL cells in hamsters and guinea pigs and only some in PRL cells of gerbils were found to be devoid of grains. Quantitative analysis revealed that the number of grains per nucleus differed considerably from cell to cell. LH cells of guinea pigs accumulated much larger amounts of3H-estradiol than did the PRL cells, while the LH cells in the hamsters and gerbils accumulated only slightly more3H-estradiol than the PRL cells. These results confirm the previous observations in rats and baboons that demonstrated tremendous species differences in percentage of cells in the anterior pituitary gland that accumulated3H-estradiol. Also, these data suggest that there are functionally heterogeneous cell types among the LH and PRL cells in hamsters, guinea pigs and gerbils as has been previously demonstrated in rats and baboons.
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