Control of photosynthate partitioning in spinach leaves

Experiments were carried out to estimate the elasticity coefficients and thence the distribution of control of sucrose synthesis and photosynthate partitioning between cytosolic fructose-1,6-bisphosphatase and sucrose-phosphate synthase (SPS), by applying the dualmodulation method of Kacser and Burns (1979, Biochem. Soc. Trans. 7, 1149–1161). Leaf discs of spinach (Spinacia oleracea L.) were harvested at the beginning and end of the photoperiod and illuminated at five different irradiances to alter (i) the extent of feedback inhibition and (ii) the rate of photosynthesis. The rate of CO₂ fixation, sucrose synthesis and starch synthesis were measured and compared with the activation of SPS, and the levels of fructose-2,6-bisphosphate (Fru2,6bisP) and metabolites. Sucrose synthesis increased progressively with increasing irradiance, accompanied by relatively large changes of SPS activity and Fru2,6bisP, and relatively small changes of metabolites. At each irradiance, leaf discs harvested at the end of the photoperiod had (compared with leaf discs harvested at the beginning of the photoperiod) a decreased rate of sucrose synthesis, increased starch synthesis, decreased SPS activity, increased Fru2,6bisP, a relatively small (20%) increase of most metabolites, no change of the glycerate-3-phosphate: triose-phosphate ratio, a small increase of NADPmalate dehydrogenase activation, but no inhibition of photosynthesis. The changes of sucrose and starch synthesis were largest in low light, while the changes of SPS and Fru2,6bisP were as large, or even larger, in high light. It is discussed how these results provide evidence that the control of sucrose synthesis is shared between SPS and fructose-1,6-bisphosphatase, and provide information about the in-vivo response of these enzymes to changes in the levels of their substrates and effectors. At low fluxes, feedback regulation is very effective at altering partitioning. In high light, changes of SPS activation and Fru2,6bisP can be readily overriden by increasing levels of metabolites.     

Regulation of circulating chromogranin B levels in heart failure

Background: Chromogranin B (CGB) regulates B-type natriuretic peptide (BNP) production. Circulating CGB levels are elevated in heart failure (HF) animal models and HF patients, but also increase in healthy individuals in response to physical activity. Therefore, CGB seems to integrate information from myocardial stress and systemic neuro-endocrine activation. Substantial gaps remain in our understanding of CGB regulation in HF.

Methods and results: We conducted a retrospective registry study including 372 patients. CGB and N-terminal pro-BNP (NT-proBNP) plasma levels were assessed in acute HF and chronic valvular HF patients and controls. CGB levels were significantly increased in acute HF and chronic valvular HF, but significantly higher in the latter. Patients in chronic valvular HF with severe mitral regurgitation (cHF-MR) showed significantly higher CGB levels than patients in chronic valvular HF with severe aortic stenosis. CGB levels progressively increased with worsening NYHA functional status and were moderately correlated to NT-proBNP, but independent of left ventricular (LV) ejection fraction (LVEF), LV mass, age and body weight. Finally, cHF-MR patients showed significant reductions of CGB levels after interventional mitral valve repair.

Conclusion: CGB is a promising emerging biomarker in HF patients with unique potential to integrate information from myocardial stress and neuro-endocrine activation.     

The influence of the parasitic angiosperm Striga gesnerioides on the growth and photosynthesis of its host, Vigna unguiculata

Cowpea plants (Vigna unguiculata) infected with the root hemiparasiticangiosperm Striga gesnerioides accumulated less biomass thanuninfected plants over a growth period of 60 d. The allometricrelationship between shoot and root dry weight was similar inparasitized plants relative to control plants, as was the proportionof dry matter partitioned into leaf, stem and root tissue. However,infected plants failed to make any significant investment ofdry matter in pods. The rate of photosynthesis of the youngestfully expanded leaf of parasitized plants was significantlylower than for control plants. The lower rates of photosynthesiswere not attributable to stomatal limitation, a loss of chlorophyllor to an accumulation of carbohydrate. The depression of photosynthesisin the young leaves was transient. As control leaves aged, photosynthesisdeclined. This also occurred in Striga infected plants, butto a lesser extent resulting in higher rates of photosynthesisin mature leaves when compared to those of uninfected plants.The foliar nitrogen content of parasitized plants was higherthan control plants consistent with the slower rate of photosyntheticdecline of older leaves. The data are discussed with respectto the influence of parasitic weeds on host growth and photosynthesis. Key words: Cowpea, hemiparasite, allometry, nitrogen     

Influence of Drought on Graminicide Phytotoxicity in Wild Oat (Avena fatua) Grown under Different Temperature and Humidity Conditions

Controlled environmental experiments were carried out to determine the phytotoxicity of several graminicides on wild oat (Avena futua L.) as influenced by combination of drought and temperature stress or drought and low relative humidity. Compared with unstressed conditions (20/15°C plus adequate soil moisture), imazamethabenz phytotoxicity to wild oat was reduced significantly when plants were exposed to a combination of drought and high temperature (30/20°C) stress. Imazamethabenz phytotoxicity was reduced almost as much by high temperature stress alone as by a combined temperature and drought stress. When herbicides were applied to wild oat plants subjected to drought alone or to drought plus high temperature, the observed reduction in phytotoxicity from greatest to least was: fenoxaprop = diclofop > flamprop > imazamethabenz. Fenoxaprop performance was most inhibited by the combination of drought plus high temperature, although drought alone and to a lesser degree, high temperature alone, inhibited fenoxaprop action. High temperature had an adverse effect on the efficacy of fenoxaprop at lower application rates. Raising fenoxaprop application rates to 400 g ha⁻¹ overcame the inhibition caused by high temperature alone but only partially alleviated the effect of drought combined with high temperature. When plants were grown under a low temperature regimen the imposition of drought stress had little effect on imazamethabenz phytotoxicity but did reduce fenoxaprop phytotoxicity. At 25/15°C drought reduced the phytotoxicity of fenoxaprop and diclofop greatly but had no significant impact on the performance of any of the herbicides examined, regardless of soil moisture regimen. Received April 14, 1997; accepted September 22, 1997     

Vitamin Q

A tentatively proposed new vitamin (Q) is based on: (1) the observation that the phospholipids of the soybean contain a factor which can substitute for platelets or the clotting factor therein, as measured by the prothrombin consumption test, and (2) the finding that, with the supplementation of the diet with these phospholipids, patients having a hereditary bleeding state characterized by a moderate thrombocytopenia, a markedly defective prothrombin consumption time and a prolonged bleeding time, show a distinct improvement in the prothrombin consumption time and a normalization of the bleeding time.     

Ribulose-1,5-bisphosphate carboxylase-oxygenase,other Calvin-cycle enzymes,and chlorophyll decrease when glucose is supplied to mature spinach leaves via the transpiration stream

The inhibition of photosynthesis after supplying glucose to detached leaves of spinach (Spinacia oleracea L.) was used as a model system to search for mechanisms which potentially contribute to the sink regulation of photosynthesis. Detached leaves were supplied with 50 mM glucose or water for 7 d through the transpiration stream, holding the leaves in low irradiance (16 mol photons · m^–2 · s^–1) and a cycle of 9 h light/15 h darkness to prevent any endogenous accumulation of carbohydrate. Leaves supplied with water only showed marginal changes of photosynthesis, respiration, enzyme levels or metabolites. When leaves were supplied with 50 mM glucose, photosynthesis was gradually inhibited over several days. The inhibition was most marked when photosynthesis was measured in saturating irradiance and ambient CO₂, less marked in saturating irradiance and saturating CO₂, and least marked in limiting irradiance. There was a gradual loss of ribulose-1,5-bisphosphate carboxylase-oxygenase (Rubisco) protein, fructose-1,6-bisphosphatase, NADP-glyceraldehyde-3-phosphate dehydrogenase and chlorophyll. The inhibition of photosynthesis was accompanied by a large decrease of glycerate-3-phosphate, an increase of triose-phosphates and fructose-1,6-bisphospate, and a small decrease of ribulose-1,5-bisphosphate. The stromal NADPH/NADP ratio increased (as indicated by increased activation of NADP-malate dehydrogenase), and the ATP/ADP ratio increased. Chlorophyll-fluorescence analysis indicated that thylakoid energisation was increased, and that the acceptor side of photosystem II was more reduced. Similar results were obtained when glucose was supplied by floating leaf discs in low irradiance on glucose solution, and when detached spinach leaves were held in high light to produce an endogenous accumulation of carbohydrate. Feeding glucose also led to an increased rate of respiration. This was not accompanied by any changes of pyruvate kinase, phosphofructokinase, or pyrophosphate: fructose-6-phosphate phosphotransferase activity. There was a decrease of phosphoenolpyruvate, glycerate-3-phosphate and glycerate-2-phosphate, an increase of pyruvate and triose-phosphates, and an increased ATP/ADP ratio. These results show (i) that accumulation of carbohydrate can inhibit photosynthesis via a long-term mechanism involving a decrease of Rubisco and other Calvin-cycle enzymes and (ii) that respiration is stimulated due to an unknown mechanism, which increases the utilisation of phosphoenolpyruvate.Abbreviations and Symbols C_i CO₂ concentration in the air space within the leaf - F_m fluorescence yield with a saturating pulse in dark-adapted material - F_o ground level of fluorescence using a weak non-actinic modulated beam in the dark - Fru1,6bisP fructose-1,6-bisphosphate - Fru1,6Pase fructose-1,6-bisphosphatase - Fru2,6bisP fructose-2,6-bisphosphate - IRGA infrared gas analyser - NAD-MDH NAD-dependent malate dehydrogenase - NADP-MDH NADP-dependent malate dehydrogenase - NADP-GAPDH NADP-dependent glyceraldehyde-3-phosphate dehydrogenase - PEP phosphoenolpyruvate - PFK phospho-fructokinase - PFP pyrophospate: fructose-6-phosphate-phosphotransferase - 3-PGA glycerate-3-phospate - P_i inorganic phosphate - Ru1,5bisP ribulose 1,5-bisphosphate - Rubisco ribulose-1,5-bisphosphate carboxylase-oxygenase - triose-phosphates sum of glyceraldehyde-3-phosphate and dihydroxyacetone phosphate This research was supported by the Deutsche Forschungsgemeinschaft (SFB 137).     

Ontogeny of two vitamin A-metabolizing enzymes and two retinol-binding proteins present in the small intestine of the rat.

The patterns of expression of cellular retinol-binding protein (CRBP), cellular retinol-binding protein, type two [CRBP(II)], lecithin: retinol acyltransferase (LRAT), and microsomal retinal reductase were examined for rat small intestine during the perinatal period. CRBP was present (15 pmole per mg soluble protein) at the earliest time examined, the 16th day of gestation, declining by 70% by birth, maintained to adulthood. In contrast, CRBP(II) appeared 2-3 days before birth, rising to its highest level (500 pmole per mg soluble protein) by day 3 after birth, then declining by 50% during the late suckling period to the adult level. Immunohistochemistry revealed that CRBP(II) initially appeared in the epithelial cell layer in a patchy manner, resolving by birth into an even staining of all villus-associated enterocytes. In contrast, CRBP was evenly expressed in the epithelial cell layer at day 17/18 but was absent by birth. Intestinal LRAT activity increased rapidly in the 2 days prior to birth, then declined at weaning to the adult level. Microsomal retinal reductase was measurable in the intestine at birth, but not detected during the early suckling period, reappearing at day 21. Considerable increase was then observed coincident with weaning, when carotenes, from which retinal is derived, became an important source of vitamin A. The pattern of appearance of these elements appears to prepare the intestine for the necessary processing of vitamin A required after birth.     

The impact of decreased Rubisco on photosynthesis, growth, allocation and storage in tobacco plants which have been transformed with antisense rbcS

Transgenic tobacco plants tranformed with antisense to rbcS to decrease expression of ribulose-1,5–bisphosphate carboxylase-oxygenase (Rubisco) have been used to investigate (a) whether Rubisco is limiting for photosynthesis and plant growth and (b) whether biomass allocation and storage of carbohydrate and nitrogen are regulated in response to decreased rate of photosynthesis. The rate of photosynthesis (measured in growth conditions) and plant growth were not strongly inhibited until almost half of the Rubisco was removed. When Rubisco was decreased further there was a large decrease of photosynthesis and plant growth. When photosynthesis decreased in the ‘antisense’ plants there was an increase in the shoot/root ratio and the specific leaf area. As a result, the leaf area ratio (leaf area per g plant dry weight) increased 3–4–fold. This shows that tobacco compensates for decreased photosynthesis by maximizing leaf area. The decrease of photosynthesis also resulted in lower starch and free hexose in the leaf, but the volume of the diurnal starch turnover was largely maintained. This indicates that partitioning to starch is regulated to decrease non-productive accumulation of starch, but still maintain a pool of transient starch for export during the night. The decrease of photosynthesis was also accompanied by a large increase of the nitrogen/ carbon balance, due to a large accumulation of nitrate in the leaf. This shows that assimilation of nitrate is inhibited in response to low availability of photo-synthate.     

Properties of soluble rat brain histone lysine methyltransferase.

Histone-lysine methyltransferase has been solubilized from rat brain chromatin by repeated extraction with distilled water. The enzyme was further purified by chromatography on DEAE-cellulose and gel filtration. With chromosomal-bound histones as substrates, the enzyme methylated only the lysyl residues in histones H3 and H4. The ratio of N epsilon-mono-: N epsilon-di-: N epsilon-trimethyllysine in histone H3 was 1.8:1.0:0.45 and the ratio of N epsilon-mono-: N epsilon-dimethyllysine in histone H4 was 0.7:1.0. The enzyme loses specificity with soluble histones as substrates; however, histones H3 and H4 were still the best methyl acceptors. The pH optima for the enzyme with soluble histones H3 and H4 as substrates were 8.2 to 8.7 and 7.2 to 8.0, respectively. S-Adenosyl-L-homocysteine, one of the products of the reaction, was a competitive inhibitor with respect to S-adenosyl-L-methionine.