Effects of ocean acidification on growth and physiology of Ulva lactuca (Chlorophyta) in a rockpool‐scenario

Rising atmospheric CO₂‐concentrations will have severe consequences for a variety of biological processes. We investigated the responses of the green alga Ulva lactuca (Linnaeus) to rising CO₂‐concentrations in a rockpool scenario. U. lactuca was cultured under aeration with air containing either preindustrial pCO₂ (280 μatm) or the pCO₂ predicted by the end of the 21st century (700 μatm) for 31 days. We addressed the following question: Will elevated CO₂‐concentrations affect photosynthesis (net photosynthesis, maximum relative electron transport rate (rETR(max)), maximum quantum yield (Fv/Fm), pigment composition) and growth of U. lactuca in rockpools with limited water exchange? Two phases of the experiment were distinguished: In the initial phase (day 1–4) the Seawater Carbonate System (SWCS) of the culture medium could be adjusted to the selected atmospheric pCO₂ condition by continuous aeration with target pCO₂ values. In the second phase (day 4–31) the SWCS was largely determined by the metabolism of the growing U. lactuca biomass. In the initial phase, Fv/Fm and rETR(max) were only slightly elevated at high CO₂‐concentrations, whereas growth was significantly enhanced. After 31 days the Chl a content of the thalli was significantly lower under future conditions and the photosynthesis of thalli grown under preindustrial conditions was not dependent on external carbonic anhydrase. Biomass increased significantly at high CO₂‐concentrations. At low CO₂‐concentrations most adult thalli disintegrated between day 14 and 21, whereas at high CO₂‐concentrations most thalli remained integer until day 31. Thallus disintegration at low CO₂‐concentrations was mirrored by a drastic decline in seawater dissolved inorganic carbon and HCO₃^?. Accordingly, the SWCS differed significantly between the treatments. Our results indicated a slight enhancement of photosynthetic performance and significantly elevated growth of U. lactuca at future CO₂‐concentrations. The accelerated thallus disintegration at high CO₂‐concentrations under conditions of limited water exchange indicates additional CO₂ effects on the life cycle of U. lactuca when living in rockpools.     

Optimization of the sucrose and ion concentrations for saikosaponin production in hairy root culture ofBupleurum falcatum

Saikosaponin productivity was examined in aBupleurum falcatum L. BFHR2 hairy root culture in response to changes in the sucrose content (2≈8%), nitrogen content (0≈250 mM NH₄NO₃), phosphate content (0≈12 mM NaH₂PO₄), and the potassium content (0≈87.2 mM KCl) of the culture media. We found that the conditions for maximal saikosaponin production differed from those for optimal root growth. Highest saikosaponin yield was achieved for 8% sucrose, 62 mM NH₄NO₃, 1.2 mM NaH₂PO₄, and 0.5 mM KCl.     

Phosphite accelerates programmed cell death in phosphate-starved oilseed rape (<Emphasis Type="Italic">Brassica napus</Emphasis>) suspension cell cultures

Phosphite (H₂PO₃^–, Phi) prevents the acclimation of plants and yeast to orthophosphate (Pi, HPO₄^2–) deprivation by specifically obstructing the derepression of genes encoding proteins characteristic of their Pi-starvation response. In this study, we report that prolonged (i.e., 3–4 weeks) culture of Brassica napus L. suspension cells in Pi-deficient (–Pi) media leads to programmed cell death (PCD). However, when the B. napus cells were subcultured into –Pi media containing 2 mM Phi, they initiated PCD within 5 days, with 95% cell death observed by day 9. Dying cells exhibited several morphological and biochemical features characteristic of PCD, including protoplast shrinkage, chromatin condensation, and fragmentation of nuclear DNA. Immunoblotting indicated that B. napus cells undergoing PCD upregulated a 30-kDa cysteine endoprotease that is induced during PCD in the inner integument cells of developing B. napus seeds. It is concluded that PCD in B. napus suspension cells is triggered by extended Pi starvation, and that Phi treatment greatly accelerates this process. Our results also infer that the adaptive value of acclimating at the molecular level to Pi-stress is to extend the viability of –Pi B. napus cell cultures by about 3 weeks.Abbreviations APase acid phosphatase (EC 3.1.3.2) - BnCysP B. napus cysteine proteinase - DAPI 4,6-diamidino-2-phenylindole - FDA fluorescein diacetate - PCD programmed cell death - Phi phosphite - +Pi and –Pi Pi-sufficient and -deficient, respectively - PI propidium iodide - PSI Pi-starvation inducible     

Growth response of komatsuna (Brassica rapa var. peruviridis) to root and foliar applications of phosphite

Soil and hydroponic culture experiments were conducted to investigate the effects of phosphite (Phi) as phosphorus (P) fertilizer via root and foliar applications on the growth and P supply of komatsuna. In both experiments, root P treatments were combinations of Phi and phosphate (Pi) at different Pi:Phi ratios, for a total of high P level (92 mg P pot^?1; the soil experiment) or low P level (0.05 mM P; the hydroponic experiment). Foliar P treatments were deionized water (control), a Pi solution and a Phi solution at low concentration of 0.05% P₂O₅. In both experiments, shoot dry weight of plants significantly decreased as Pi:Phi ratio decreased. In the soil experiment, plants grew abnormally at a Pi:Phi ratio of 25:75 and died when P was applied to soil entirely as Phi form (0:100 treatment). In the hydroponic experiment, no visible damage was found in shoot but root growth was strongly inhibited with severe damage symptoms at low Pi:Phi ratios. Total P concentration in plant decreased significantly with decreasing Pi:Phi ratio, especially in the hydroponic experiment. Foliar application of Phi although greatly increased total P of plants compared to that of Pi in both experiments, it did not improve but further decreased plant growth at low Pi:Phi ratios in the soil experiment and at all Pi:Phi ratios in the hydroponic experiment. The results of this study clearly indicated that Phi could not be used as P fertilizer by komatsuna plants via both application methods and could not substitute P at any rate at either low or high level. No beneficial effect of Phi was detected even when it was applied at low rate or applied in combination with Pi at different ratios. The effects of Phi were strongly dependent on the P nutrition status of plants; and plants that were not sufficiently fertilized with Pi may become vulnerable to Phi even at low levels.     

PHOSPHATE UPTAKE KINETICS IN EUGLENA GRACILIS (Z) (EUGLENOPHYCEAE) GROWN ON LIGHT/DARK CYCLES.' I. SYNCHRONIZED BATCH CULTURES1

The characteristics of phosphate uptake in synchronized populations of Euglena gracilis Klebs (Z) were studied. The cells were grown autotrophically in batch culture and synchronized with a cycle of 14:10 LD. Incorporation of P was nonlinear with time for the first 2 h of incubation over a wide range of P concentrations and completely inhibited by darkness. The kinetics of P uptake as a function of P concentration were triphasic between 0 and 100 μM PO₄, obeying Michaelis-Menten kinetics over the 0–3 μM PO₄ range-only. Uptake velocity increased linearly with, concentration above 3 μM PO₄. The kinetics of P uptake varied with stage in the cell cycle. The half-saturation constant for uptake at the lower concentrations oscillated between 0.7 and 2.8 μM PO₄, reaching a peak immediately before the onset of cell division (beginning of the dark period). V_max was largest in the middle of the light period, as was the slope of the linear portion of the kinetic pattern. Further analysis of the kinetics suggests that changes in this slope are responsible for the oscillation in K_s values calculated for the lower concentrations. This analysis assumes 2 uptake mechanisms, one which saturates at low concentrations of phosphate, and one which is nonsaturable over the entire concentration range examined.     

PHOTOSYNTHETIC PROPERTIES OF PROTOPLASTS,AS COMPARED WITH THALLI,OF ULVA FASCIATA (CHLOROPHYTA)1

Protoplasts were prepared from Ulva fasciata Delile, and their photosynthetic performance was measured and compared with that of thalli discs. These protoplasts maintained maximal rates of photosynthesis as high as those of thalli (up to 300 μmol O₂·mg chlorophyll^?1·h^?1) for several hours after preparation and were therefore considered suitable for kinetic studies of inorganic carbon utilization. The photosynthetic K_1/2(inorganic carbon) at pH 6.1 was 3.8 μM and increased to 67, 158, and 1410 μM at the pH values 7.0, 7.9, and 8.9, respectively. Compared with these protoplasts, thalli had a much lower affinity for CO₂ but approximately the same affinity for HCO₃^?. Comparisons between rates of photosynthesis and the spontaneous dehydration of HCO₃^? (at 50 μM inorganic carbon) revealed that photosynthesis of both protoplasts (which lacked apparent activity of extracellular/surface-bound carbonic anhydrase) and thalli (which were only 25% inhibited by the external carbonic anhydrase inhibitor acetazolamide) could not be supported by CO₂ formation in the medium at the higher pH values, indicating HCO₃^? uptake. Since both protoplasts and thalli were sensitive to 4,4′-diisothiocyanostilbene-2,2′-disulfonate, we suggest that HCO₃^? transport was facilitated by the membrane-located anion exchange protein recently reported to function in certain Ulva thalli. These findings suggest that the presence of a cell wall may constitute a diffusion barrier for CO₂, but not for HCO₃^?, utilization under natural seawater conditions.     

Disruption of the phosphate-starvation response of oilseed rape suspension cells by the fungicide phosphonate

The influence of the anti-fungal agent phosphonate (Phi) on the response of oilseed rape (Brassica napus L. cv. Jet Neuf ) cell suspensions to inorganic phosphate (Pi) starvation was examined. Subculture of the cells for 7 d in the absence of Pi increased acid phosphatase (APase; EC 3.1.3.2) and pyrophosphate (PPi)-dependent phosphofructokinase (PFP; EC 2.7.1.90) activities by 4.5- and 2.8-fold, respectively, and led to a 19-fold increase in V _max and a 14-fold decrease in K _m (Pi) for Pi uptake. Addition of 2 mM Phi to the nutrient media caused dramatic reductions in the growth and Pi content of the Pi-starved, but not Pi-sufficient cells, and largely abolished the Pi-starvation-dependent induction of PFP, APase, and the high-affinity plasmalemma Pi translocator. Immunoblotting indicated the cells contain three APase isoforms that are synthesized de novo following Pi stress, and that Phi treatment represses this process. Phosphonate treatment of Pi-starved cells significantly altered the relative extent of in-vivo ³²P-labelling of polypeptides having M_rs of 66, 55, 45 and 40 kDa. However, Phi had no effect on the total adenylate pool of Pi-starved cells which was about 32% lower than that of Pi-sufficient cells by day 7. Soluble protein levels, and activities of pyruvate kinase (EC 2.7.1.40) and ATP-dependent phosphofructokinase (EC 2.7.1.11) were unaffected by Pi starvation and/or Phi treatment. The effects of Phi on the growth, and APase and PFP activities of Pi-starved B. napus seedlings were similar to those observed in the suspension cells. The results are consistent with the hypothesis that a primary site of Phi action in higher plants is at the level of the signal transduction chain by which plants perceive and respond to Pi stress at the molecular level. Received: 30 December 1996 / Accepted: 19 February 1997     

Growth characteristics of two bloom‐forming synurophytes (Mallomonas elongata and Synura petersenii) at different nitrate and phosphate concentrations

Two dominant planktonic bloom‐forming algal species in a small shallow eutrophic pond were identified as Mallomonas elongata and Synura petersenii by electron microscopy. Their growth requirements were investigated as uni‐algal cultures in a laboratory study. The maximum population growth and maximum growth rate of M. elongata occurred at concentrations of 24 μM nitrate (NO₃) and 5 μM phosphate (PO₄) at a temperature of 15°C and a pH of 6. Synura petersenii grew maximally and exhibited the highest growth rate at a NO₃ concentration of 24 μM and a PO₄ concentration of 2 μM. Mallomonas elongata and S. petersenii had similar nutrient requirements for optimum growth, suggesting that the biomass of these two species can be controlled by nutrient gradients.     

Phosphorylcholine: a specific promoter of reparin and serum -lipoprotein interaction

The mechanism of formation of heparin-β-lipoprotein complex was studied. Precipitation of fatty acid labelled β-lipoproteins and optical density changes of the reaction were used as measures for the formation of insoluble complex. Equimolar amounts of Ca-phosphorylcholine were found to be more effective than CaCl₂ to form the insoluble complex. In presence of 10 mM CaCl₂, minute amounts (2–16 μM) of Ca or Na-phosphorylcholine increased the interaction between heparin and serum β-lipoprotein. Equimolar amounts of choline, ethanolamine, acetylcholine, phosphoserine, phosphothreonine, α-glycerophosphate, α-glycerophosphate plus choline, NaH₂PO₄, galactose, glucosamine and N-acetylglucosamine had no effect on the reaction. The results suggested that phosphorylated quaternary nitrogen groups play a role in the interaction of β-lipoprotein with heparin and divalent cation.     

LIPID CLASS,CAROTENOID, AND TOXIN DYNAMICS OF KARENIA BREVIS (DINOPHYCEAE) DURING DIEL VERTICAL MIGRATION1

The internal lipid, carotenoid, and toxin concentrations of Karenia brevis (C. C. Davis) Gert Hansen and Moestrup are influenced by its ability to use ambient light and nutrients for growth and reproduction. This study investigated changes in K. brevis toxicity, lipid class, and carotenoid concentrations in low‐light, nitrate‐replete (250 μmol quanta · m^?2 · s^?1, 80 μM NO₃); high‐light, nitrate‐replete (960 μmol quanta · m^?2 · s^?1, 80 μM NO₃); and high‐light, nitrate‐reduced (960 μmol quanta · m^?2 · s^?1, <5 μM NO₃) mesocosms. Reverse‐phase HPLC quantified the epoxidation state (EPS) of the xanthophyll‐cycle pigments diadinoxanthin and diatoxanthin, and a Chromarod Iatroscan thin layer chromatography/flame ionization detection (TLC/FID) system quantified changes in lipid class concentrations. EPS did not exceed 0.20 in the low‐light mesocosm, but increased to 0.65 in the high‐light mesocosms. Triacylglycerol and monogalactosyldiacylglycerol (MGDG) were the largest lipid classes consisting of 9.3% to 48.7% and 37.3% to 69.7% of total lipid, respectively. Both lipid classes also experienced the greatest concentration changes in high‐light experiments. K. brevis increased EPS and toxin concentrations while decreasing its lipid concentrations under high light. K. brevis may mobilize its toxins into the surrounding environment by reducing lipid concentrations, such as sterols, limiting competition, or toxins are released because lipids are decreased in high light, reducing any protective mechanism against their own toxins.     

LUXURY PHOSPHATE UPTAKE AND VARIATION OF INTRACELLULAR METAL CONCENTRATIONS IN HETEROSIGMA AKASHIWO (RAPHIDOPHYCEAE)1

The effectr of phosphate starvation and subsequent uptake on distribution and concentration of phosphate metabolic intermediates and metals were studied in Heterosigma akashiwo (Hada) Hada by ³¹P-NMR spectroscopy, neutron activation analysis and ESR spectroscopy. Excess orthophosphate (4.5 μM P_i, as NaH₂PO₄) added to a medium with P-depleted H. akashiwo cells was rapidly taken up resulting in an increase in P cell quota (q_p)from 68.2 to 99.6 fmol. cell-¹in 2 h and to 156.3 fmol. cell-¹in 6 h. After three days, q_p approached about 190 fmol. cell^?1. Polyphosphate (PP_i) rapidly increased from 0 to 11.4 fmol· cell^?1in 2 h and to 24.7 fmol·cell^?1in 6 h. Diel variation of cell quota indicated that cellular P_i increase was synchronized with cellular PP_i decrease and vice versa. The average chain length of PP_i increased from ca. 0 to ca. 10.2 phosphate residues in 2 h after addition of P_i and one day later, from ca. 9.8 to ca. 12.5. The cell quota of Mn (q_Mn), and to a lesser extent Co, increased rapidly from 4.87 fg. cell^?1in the P- starved condition to 50.48 fg·cell^?12 h afer addition of P_i but decreased to 8.63 fg. Cell^?1by 6 h. Concentrations of Zn, As, Hf, Cu and sometimes Al, Mg, K, and Ca changed in a manner opposite to that of Mn and Co. The excretion of these cations, which was synchronized with the uptake of Mn and Co, may be important for a charge balancing in the cells. The ESR spectra showed that the high cellular Mn observed at 2 h after P addition was Mn²⁺which was taken up by the cells rather than adsorbed on the cell surface. These data combined with PP_i data suggested that the behavior of q_Mn is synchronized with the behavior of average chain length of PP_i.     

The Influence of Light and Oxygen on Nitrogenase Activity in the Lichen Stereocaulon paschale

Thalli of the lichen Slereocaulon paschale (L.) Fr. were prctreated in the light (light activated) or in the dark (dark starved). In short-time experiments with both light activated arid dark starved thalli, the nitrogenasc activity was higher in the light than in the dark, Light activated thalli had a very much higher rale of C₂H₂ reduction than dark starved thalli, both in the light and in the dark. The dark starved lhalli showed increasing nilrogenase activity when incubated in the light. Either light or oxygen was necessary for nitrogenase activity in light activated thalli. and up to about 10kPa oxygen they showed additive effects. Both in the light and in the dark the nitrogenase activity decreased when the oxygen partial pressure was lower than in normal air. The experimental data thus showed a short-term effect of light on nilrogenase activity by provision of ATP and reductant, and a long term effect probably by build up of reserves that were later utilized. Any immediate effect of photorespiration on nitrogenase activity could not be found in light activated thalli.     

DECREASED RATE OF GLUCOSE UTILIZATION BY RAT BRAIN IN VIVO AFTER EXPOSURE TO ATMOSPHERES CONTAINING HIGH CONCENTRATIONS OF CO2

—(1) The effects of exposure of rats to increased atmospheric concentrations of CO₂ on brain metabolism in vivo were studied. (2) After 2·5 min exposure to an atmosphere of 20% CO₂, the rate of glucose utilization by brain decreased from 0·61 μmol/min per g to 0·32 μmol/min per g and remained between 0·3 and 0·4 μmol/min per g for 60 min, the longest interval studied. O₂ utilization, calculated from the arteriovenous difference of O₂ across the brain and blood flow, was 3·5 μmol/min per g in controls and was 4·7 μmol/min per g after 5 min in the 20% CO₂ atmosphere. (3) The concentrations of glucose, glucose 6-phosphate and aspartate were increased during the first 10 min of CO₂ exposure whereas the concentrations of other glycolytic intermediates, tricarboxylic acid cycle intermediates and glutamate were decreased. The amount of endogenous substrate which disappeared during the first 10 min was sufficient, if used to supplement glucose as a fuel, to maintain the O₂ consumption at, or slightly above, the control level. Glutamate and lactate were quantitatively the most important energy sources. (4) The mechanism whereby‘CO₂ decreased the rate of glucose utilization is uncertain. The initial rise in glucose 6-phosphate and fall in fructose 1,6-diphosphate concentrations suggested that an inhibition of phosphofructokinase was responsible. However, after 60 min in 20% CO₂, the concentrations of both of these metabolites returned to normal while the rate of glucose utilization remained depressed.     

Nonformaldehyde durable press finishing of cotton fabrics using the combination of maleic acid and sodium hypophosphite

Polycarboxylic acids have been used as nonformaldehyde crosslinking agents for cotton with sodium hypophosphite (NaH₂PO₂) as the catalyst to replace the formaldehyde-based dimethyloldihydroxyethleneurea (DMDHEU). Maleic acid (MA), an α, β-unsaturated bifunctional carboxylic acid, can esterify cotton but is not able to form crosslinking between two cellulose molecules by itself. In this research, we discovered that the wrinkle resistance of the cotton fabric treated with MA and NaH₂PO₂ was significantly increased and phosphorus was bound to cotton when the treated fabric was exposed to temperatures higher than that required for esterification of cotton by MA. Elevation of the fabric wrinkle resistance and increase in quantity of the phosphorus bound to cotton had similar dependency on curing temperature, on MA concentration, and on NaH₂PO₂ concentration. All the data support the hypothesis that H-P-(residual of NaH₂PO₂) added to >CC< of the MA already bound to cotton by esterification, thus forming a new crosslink between two cotton cellulose molecules. The cotton fabrics treated by MA/NaH₂PO₂ showed fabric wrinkle resistance similar to that treated with DMDHEU, but the breaking strength and tearing strength of the MA-treated cotton fabrics were significantly improved.     

Incorporation of extracellular fatty acids by a fatty acid kinase‐dependent pathway in Staphylococcus aureus

Acyl‐CoA and acyl‐acyl carrier protein (ACP) synthetases activate exogenous fatty acids for incorporation into phospholipids in Gram‐negative bacteria. However, Gram‐positive bacteria utilize an acyltransferase pathway for the biogenesis of phosphatidic acid that begins with the acylation of sn‐glycerol‐3‐phosphate by PlsY using an acyl‐phosphate (acyl‐PO₄) intermediate. PlsX generates acyl‐PO₄ from the acyl‐ACP end‐products of fatty acid synthesis. The plsX gene of Staphylococcus aureus was inactivated and the resulting strain was both a fatty acid auxotroph and required de novo fatty acid synthesis for growth. Exogenous fatty acids were only incorporated into the 1‐position and endogenous acyl groups were channeled into the 2‐position of the phospholipids in strain PDJ39 (ΔplsX). Extracellular fatty acids were not elongated. Removal of the exogenous fatty acid supplement led to the rapid accumulation of intracellular acyl‐ACP and the abrupt cessation of fatty acid synthesis. Extracts from the ΔplsX strain exhibited an ATP‐dependent fatty acid kinase activity, and the acyl‐PO₄ was converted to acyl‐ACP when purified PlsX is added. These data reveal the existence of a novel fatty acid kinase pathway for the incorporation of exogenous fatty acids into S. aureus phospholipids.     

Effects of NaCI stress on nitrogen and phosphorous metabolism in a true mangrove Bruguiera parviflora grown under hydroponic culture

The influence of varying levels of salinity (0, 100, 200 and 400 mM) on the activities of nitrate reductase (NR, E.C. 1.6.6.1), acid phosphatase (ACP, E.C. 3.1.3.2), and alkaline phosphatase (ALP, EC 3.1. 3.1) as well as on nitrate and phosphate uptake and total nitrogen levels in leaves of a true mangrove Bruguiera parviflora was investigated under hydroponic culture conditions. NR activity increased in 100 mM NaCl treated plants, whereas it decreased gradually in 200 and 400 mM treated plants, relative to the controls. Decreased activity of NR by NaCl stress was also accompanied by a decrease in total nitrogen level and nitrate uptake. Decreases in NR activity, nitrate (NO₃⁻), and total nitrogen level due to high salinity may be responsible for a decrease in growth and biomass production in this plant. However, salinity caused an increase in both ACP and ALP activity. Activity staining of ACP by native polyacrylamide gel electrophoresis revealed three isoforms: ACP-1, ACP-2, and ACP-3. We observed a preferential enhancement in the ACP-3 isoform by salinity. In order to understand whether the salinity-induced increase in phosphatase activity was due to inhibition in phosphate uptake, we monitored phosphate (Pi) levels in leaves and noted that phosphate levels decreased significantly under salinity. These results suggest that the induction of acid and ALP under salt stress may be due to a phosphorous deficiency.     

Characterization of phosphate absorption in maize root cortex segments

Uptake of phosphate ions by 1 mm segments of isolated maize root cortex layers was studied. Cortex segments (from roots of 8 days old maize plants) absorb phosphate ions from 1 mM KH₂PO₄ in 0.2 mM CaSCO₄ at the average rate of 34.3 ±3.2 μg P_i g^?1 (fr. m.) h^?1,i.e. 0.35± 0.02 μmol P_i g^?1 (fr. m.) h^?1. Phosphate uptake considerably increases after a certain period of “augmentation”,i.e. washing in aerated 0.2 mM CaSO₄. This increase is completely blocked by the presence of 10 μg ml^?1 cycloheximide. The relation of uptake rate to phosphate concentration in the medium was shown to have 3 phases in the concentration range of 0.02 - 40 mM. Transition points were found between 0.8–1 mM and 10–20 mM. Following K_m and V_max values were found: K_m[mM] : 0.37 - 3.82 - 27.67 V_max[μg P_i g^?1 (fr. m.) h^?1] : 3.33 - 39.40 - 66.67 We have found no sharp pH optimum for phosphate uptake. It proceeds at almost constant rate till pH 6.0 and then the uptake rate drops with increasing pH. At low phosphate concentrations (1 mM) the lowest uptake rate was found at 5 and 13 °C, while the uptake is higher at 5 °C than at 13 °C at phosphate concentrations higher than 1 mM. At these concentrations uptake rate at 35 °C is lower than at 25 °C. Phosphate uptake considerably decreased in anaerobic conditions. DNP and iodoacetate (0.1 mM) completely blocked phosphate uptake from 1 mM KH₂PO₄, while uptake from 5 and 10 mM KH₂PO₄ was left unaffected by these substances. The inhibitors of active - SH groups NEM and PCMB inhibited phosphate uptake: 10^?3 M NEM by 81.6%, 10⁴ M NEM by 42% and 10^?4 M PCMB by 42%.     

Growth,nutrient dynamics,and efficiency responses to carbon dioxide and phosphorus nutrition in soybean

Plant mineral nutrients such as phosphorus may exert major control on crop responses to the rising atmospheric carbon dioxide (CO₂) concentrations. To evaluate the growth, nutrient dynamics, and efficiency responses to CO₂ and phosphorus nutrition, soybean (Glycine max (L.) Merr.) was grown in controlled environment growth chambers with sufficient (0.50 mM) and deficient (0.10 and 0.01 mM) phosphate (Pi) supply under ambient and elevated CO₂ (aCO₂, 400 and eCO₂, 800 µmol mol^?1, respectively). The CO₂ × Pi interaction was detected for leaf area, leaf and stem dry weight, and total plant biomass. The severe decrease in plant biomass in Pi-deficient plants (10–76%) was associated with reduced leaf area and photosynthesis (P_net). The degree of growth stimulation (0–55% total biomass) by eCO₂ was dependent upon the severity of Pi deficiency and was closely associated with the increased phosphorus utilization efficiency. With the exception of leaf and root biomass, Pi deficiency decreased the biomass partitioning to other plant organs with the maximum decrease observed in seed weight (8–42%) across CO₂ levels. The increased tissue nitrogen (N) concentration in Pi-deficient plants was accredited to the lower biomass and increased nutrient uptake due to the larger root to shoot ratio. The tissue P and N concentration tended to be lower at eCO₂ versus aCO₂ and did not appear to be the main cause of the lack of CO₂ response of growth and P_net under severe Pi deficiency. The leaf N/P ratio of >16 was detrimental to soybean growth. The tissue P concentration needed to attain the maximum productivity for biomass and seed yield tended to be higher at eCO₂ versus aCO₂. Therefore, the eCO₂ is likely to increase the leaf critical P concentration for maximum biomass productivity and yield in soybean.     

The effects of methionine and cobalt ions on sex expression and development of <Emphasis Type="Italic">Anemia phyllitidis</Emphasis> gametophytes

One of the prime precursor for ethylene synthesis — L-methionine and the inhibitor of 1-aminocyclopropane-1-carboxylic acid oxidase (ACO) — Co²⁺-were tested for their effects on sex expression and development of Anemia phyllitidis fern gametophytes. Five concentrations of both chemicals (0, 10, 25, 50, 100 μM) were analysed with reference to antheridia and archegonia formation, number and size of cells as well as thalli length using the three-zone model of gametophyte structure. Both substances, however at different concentrations, enhanced the number of GA₃-induced antheridia and similarly stimulated the cell number and inhibited thalli length. Both of them at 100 μM concentrations without GA₃ induced meristematic area formation while methionine also induced archegonia in the apical parts of gametophytes. These findings correspond with the previous observations concerning the important role of ethylene synthesis precursor (ACC) in controlling gibberellic acid-induced male sex expression in ferns and broaden the knowledge about the mechanisms of fern gametophyte development.