Effects of pH and nitrogen on cadmium uptake in potato

This study investigated the effects of pH and nitrogen form and concentration on cadmium (Cd) uptake by potato (Solanum tuberosum L.) grown in hydroponic culture. Potato plants grown in a pH-buffered nutrient solution for 10 d were exposed for 24 h to 25 nM CdCl₂ labelled with ¹⁰⁹Cd. Plants showed a significantly higher Cd uptake and accumulation at pH 6.5 than at pH 4.5 and 5.5. Nitrogen supplied as nitrate (NO₃ ^?) generally resulted in a higher Cd uptake and accumulation than N supplied as ammonium (NH₄ ⁺). This effect was most pronounced at pH 6.5. The N concentration increasing from 6.5 to 26 mM resulted in a decreased Cd influx when either NO₃ ^? or NH₄ ⁺ was used. Cd translocation to the shoot was increased when NO₃ ^? was used as the sole N source. In conclusion, pH had a strong influence on Cd uptake by roots and N form is especially important for Cd translocation within the potato plant.     

Effects of pH on ammonium uptake by Typha latifolia L.

The effects of solution pH on NH₄⁺ uptake kinetics and net H⁺ extrusion by Typha latifolia L. were studied during short-term (days) and long-term (weeks) exposure to pH in the range of pH 3.5–8.0. The NH₄⁺ uptake kinetics were estimated from depletion curves using a modified Michaelis-Menten model. T. latifolia was able to grow in solution culture with NH₄⁺ as the sole N source and to withstand a low medium pH for short periods (days). With prolonged exposure (weeks) to pH 3.5, however, the plants showed severe symptoms of stress and stopped growing. The solution pH affected NH₄⁺ uptake kinetics. The affinity for NH₄⁺, as quantified by the half saturation constant (K_1/2) and C_min (the NH₄⁺ concentration at which uptake ceases), decreased with pH. K_1/2 was increased from 7.1 to 19.2 mmol m^?3 and C_min from 2.0 to 5.7 mmol m^?3 by lowering the pH in steps from 8.0 to 3.5. V_max was, however, largely unaffected by pH (～22 μmol h^?1 g^?1 root dry weight). Under prolonged exposure to constant pH, growth rates were highest at PH 5.0 and 6.5. At pH 8.0 growth was slightly depressed and at pH 3.5 growth completely stopped. NH₄⁺ uptake kinetics were similar at pH 5.0, 6.5 and 8.0 whereas at pH 3.5 NH₄⁺ uptake almost completely stopped. The ratio between net H⁺ extrusion and NH₄⁺ uptake decreased significantly at low pH. The adverse effects of low pH on NH₄⁺ uptake kinetics are probably a consequence of a reduced H⁺-ATPase activity and/or an increased re-entry of H⁺ at low pH, and the associated decrease in the electrochemical gradient across the plasma membranes of the root cells.     

Ammonium and nitrate uptake rates and rhizosphere pH in non-mycorrhizal roots of Norway spruce [Picea abies (L.) Karst.]

Summary Relationships between root zone temperature, concentrations and uptake rates of NH ₄ ⁺ and NO ₃ ^– were studied in non-mycorrhizal roots of 4-year-old Norway spruce under controlled environmental conditions. Additionally, in a forest stand NH ₄ ⁺ and NO ₃ ^– uptake rates along the root axis and changes in the rhizosphere pH were measured. In the concentration (C_min) range of 100–150 M uptake rates of NH ₄ ⁺ were 3–4 times higher than those of NO ₃ ^– The preference for NH ₄ ⁺ uptake was also reflected in the minimum concentration (C_min) values. Supplying NH₄NO₃, the rate of NO ₃ ^– uptake was very low until the NH ₄ ⁺ concentrations had fallen below about 100 M. The shift from NH ₄ ⁺ to NO ₃ ^– uptake was correlated with a corresponding shift from net H⁺ production to net H⁺ consumption in the external solution. The uptake rates of NH ₄ ⁺ were correlated with equimolar net production of H⁺. With NO ₃ ^– nutrition net consumption of H⁺ was approximately twice as high as uptake rates of NO ₃ ^– In the forest stand the NO ₃ ^– concentration in the soil solution was more than 10 times higher than the NH ₄ ⁺ concentration (<100 M), and the rhizosphere pH of non-mycorrhizal roots considerably higher than the bulk soil pH. The rhizosphere pH increase was particularly evident in apical root zones where the rates of water and NO ₃ ^– uptake and nitrate reductase activity were also higher. The results are summarized in a model of water and nutrient transport to, and uptake by, non-mycorrhizal roots of Norway spruce in a forest stand. Model calculations indicate that delivery to the roots by mass flow may meet most of the plant demand of nitrogen and calcium, and that non-mycorrhizal root tips have the potential to take up most of the delivered nitrate and calcium.     

INHIBITION OF CARBON UPTAKE AND STIMULATION OF NITRATE UPTAKE AT LOW SALINITIES IN FUCUS DISTICHUS (PHAEOPHYCEAE)1

Inorganic carbon and nitrate uptake were examined in whole plants of Fucus distichus L. (Powell) incubated in dilutions of synthetic ocean water and media with different concentrations of Na⁺, K⁺ and Cl^?. Reduction in salinity from normal seawater (33 ppt) decreased carbon uptake rate but increased nitrate uptake rate by 50% each. Substitution of K⁺ for Na⁺ at constant ionic strength decreased nitrate uptake. Substitution of K⁺ or mannitol for Na⁺ decreased carbon uptake. Neither the uptake of nitrate or carbon was changed by substituting SO₄^2- for Cl^?. Ionophores, valinomycin and monensin, inhibited both nitrate uptake and carbon fixation from 20 to 70% of control rates. The stimulation of nitrate uptake at low salinity may be beneficial to plants in estuarine tidal environments in which nitrate is supplied by the fresh water source.     

Nutrient uptake by intact and disturbed roots of loblolly pine seedlings

Most measurements of nutrient uptake use either hydroponic systems or soil-grown roots that have been disturbed by excavation. The first objective of this study was to test how root excavation affects nitrate uptake. Rates of NO₃^? uptake by mycorrhizal loblolly pine (Pinus taeda L.) seedlings were measured in intact sand-filled columns, hydroponics, and disturbed sand-filled columns. Total nitrate uptake in intact sand-filled columns was higher than in disturbed columns, indicating that disturbance lowers uptake. Transferring plants from the sand-filled columns to hydroponics had little effect on NO₃^? uptake beyond delaying uptake for an hour. The second objective of this study was to determine whether NH₄⁺, Ca²⁺, Mg²⁺ and K⁺ uptake could be studied using sand-filled columns, since previous studies had tested this method only for nitrate uptake. Uptake rates of NH₄⁺ and K⁺ were positive, while Ca²⁺ and Mg²⁺ uptake rates were negative in intact sand-filled columns, indicating that net efflux may occur even without physical disturbance to the root system. The sand-filled column approach has some limitations, but holds promise for conducting nutrient uptake studies with minimal disturbance to the root system.     

Isolation and characterization of a <Emphasis Type="Italic">Mastigocladus</Emphasis> species capable of growth,N<Subscript>2</Subscript>-fixation and N-assimilation at elevated temperature

A Mastigocladus species was isolated from the hot spring of Jakrem (Meghalaya) India. Uptake and utilization of nitrate, nitrite, ammonium and amino acids (glutamine, asparagine, arginine, alanine) were studied in this cyanobacterium grown at different temperatures (25°C, 45°C). There was 2–3 fold increase in the heterocyst formation and nitrogenase activity in N-free medium at higher temperature (45°C). Growth and uptake and assimilation of various nitrogen sources were also 2–3 fold higher at 45°C indicating that it is a thermophile. The extent of induction and repression of nitrate uptake by NO₃ ⁻ and NH₄ ⁺, respectively, differed from that of nitrite. It appeared that Mastigocladus had two independent nitrate/nitrite transport systems. Nitrate reductase and nitrite reductase activitiy was not NO₃ ⁻-inducible and ammonium or amino acids caused only partial repression. Presence of various amino acids in the media partially repressed glutamine synthetase activity. Ammonium (methylammonium) and amino acid uptake showed a biphasic pattern, was energy-dependent and the induction of uptake required de novo protein synthesis. Ammonium transport was substrate (NH₄ ⁺)-repressible, while the amino acid uptake was substrate inducible. When grown at 25°C, the cyanobacterium formed maximum akinetes that remained viable upto 5 years under dry conditions.     

Nitrogen and base cation uptake in seedlings of Acer pseudoplatanus and Calamagrostis villosa exposed to an acidified environment

The effects of solution acidity and form of nitrogen on net nutrient uptake rates in Acer pseudoplatanus and Calamagrostis villosa seedlings were examined as part of a complex ecological study. Uptake rates were measured by the depletion method under controlled conditions (temperature 20 °C, irradiance 400 mol m^–2 s^–1 PAR) from a nutrient solution containing 1.5 mM nitrogen in the form of nitrate or ammonium or an equimolar mixture of both. The solution acidity was kept constant at pH 5.5 (control treatment), 4.5 or 3.5 (low pH treatments). Strongly acid pH decreased or stopped the uptake rates of NO₃ ^–, Mg²⁺ and Ca²⁺, but the uptake of NH₄ ⁺ was not changed in both species. Ammonium ions reduced the uptake rate of NO₃ ^– in Acer but increased the uptake rate in Calamagrostis. Ammonium as the sole source of nitrogen had a strong negative impact on the uptake rates of calcium and magnesium and this effect was independent of the media acidification usually connected with NH₄ ⁺ uptake and assimilation. However, the negative effect of ammonium ions on the base cation uptake was more pronounced at low pH values.     

Extra- and Intracellular pH and Membrane Potential Changes Induced by K, Cl, H(2)PO(4), and NO(3) Uptake and Fusicoccin in Root Hairs of Limnobium stoloniferum

Short-term ion uptake into roots of Limnobium stoloniferum was followed extracellularly with ion selective macroelectrodes. Cytosolic or vacuolar pH, together with the electrical membrane potential, was recorded with microelectrodes both located in the same young root hair. At the onset of chloride, phosphate, and nitrate uptake the membrane potential transiently decreased by 50 to 100 millivolts. During Cl⁻ and H₂PO₄⁻ uptake cytosolic pH decreased by 0.2 to 0.3 pH units. Nitrate induced cytosolic alkalinization by 0.19 pH units, indicating rapid reduction. The extracellular medium alkalinized when anion uptake exceeded K⁺ uptake. During fusicoccin-dependent plasmalemma hyperpolarization, extracellular and cytosolic pH remained rather constant. Upon K⁺ absorption, FC intensified extracellular acidification and intracellular alkalinization (from 0.31 to 0.4 pH units). In the presence of Cl⁻ FC induced intracellular acidification. Since H⁺ fluxes per se do not change the pH, recorded pH changes only result from fluxes of the stronger ions. The extra- and intracellular pH changes, together with membrane depolarization, exclude mechanisms as K⁺/A⁻ symport or HCO₃⁻/A⁻ antiport for anion uptake. Though not suitable to reveal the actual H⁺/A⁻ stoichiometry, the results are consistent with an H⁺/A⁻ cotransport mechanism.     

Effects of pH,phosphate and ammonia on the rate of uptake of nitrate and ammonia by freshwater phytoplankton

The effect of phosphate (PO₄ ⁺³) and pH in regulating nitrate (NO₃) and ammonia (NH₃ ⁺) uptake by phytoplankton was investigated in two Oklahoma lakes using ¹⁵N tracers. Addition of PO₄ ⁺³ above ambient concentrations had a negligible effect on the rate of uptake of NO₃ ^– or NH₃ ⁺. Manipulation of pH of lake water had little effect on uptake of either NO₃ ^– or NH₃ ⁺. A correlation analysis suggested that NO₃ ^– is not used by phytoplankton when NH₃ ⁺ concentrations exceed about 210 µg NH₃ ⁺-N(1)^–1.     

Effect of nickel on certain physiological and biochemical behaviors of an acid tolerantChlorella vulgaris

This study concerns the inhibitory effects of acid pH and nickel on growth, nutrient (NO₃ ^- and NH₄ ⁺) uptake, carbon fixation, O₂ evolution, electron transport chain and enzyme (nitrate reductase and ATPase) activities of acid tolerant and wild-type strains of Chlorella vulgaris. Though a general reduction in all these variables was noticed with decreasing pH, the tolerant strain was found to be metabolically more active than the wild-type. A reduced cation (NH₄ ⁺, Na⁺, K⁺ and Ca²⁺) uptake, coupled with a facilitated influx of anions (NH₄ ⁺, PO₄ ^3- and HCO₃ ^-), suggested the development of a positive membrane potential in acid tolerant Chlorella. Nevertheless, a tremendous increase in ATPase activity at decreasing pH revealed the involvement of superactive ATPase in exporting H⁺ ions and keeping the internal pH neutral. A difference in Na⁺ and K⁺ efflux of the two strains at decreasing pH suggests there is a difference in membrane permeability. The low toxicity of Ni in the acid tolerant strain may be due to the low Ni uptake brought about by a change in membrane potential as well as in permeability. Hence, the development of superactive ATPase and a change in both membrane potential and permeability not only offers protection against acidity, but also co-tolerance to metals.     

Changes in haemolymph ion concentrations of Astacus astacus L. and Pacifastacus leniusculus (Dana) after exposure to low pH and aluminium

During exposure to soft water, acidified to pH 4.0, the haemolymph concentrations of Na⁺, K⁺, and Cl^– decreased whereas the Ca²⁺ concentration fluctuated in Astacus astacus. The haemocyte content of K⁺ decreased from 9% to 2% of the total haemolymph K⁺ content after exposure to pH 3.7 for 3 days. Within 14 days, 250 µg Al³⁺ l^–1, as Al₂(SO₄)₃ at pH 5.0, reduced the haemolymph Na⁺ content in Astacus astacus and Pacifastacus leniusculus, however, the effects were less pronounced than earlier reported for fish. Disturbed ion regulation, mainly depending on low pH, is thought to contribute to the absence of these species in acid waters.     

Decrease in nitrate uptake and increase in proton release in zinc deficient cotton,sunflower and buckwheat plants

The effect of varied Zn supply on the pH of the nutrient solution and uptake of cations and anions was studied in cotton (Gossypium hirsutum L.), sunflower (Helianthus annuus L.) and buckwheat (Fagopyrum esculentum Moench) plants grown under controlled environmental conditions in nutrient solutions with nitrate as source of nitrogen. With the appearance of visual Zn deficiency symtoms, the pH of the nutrient solutions decreased from 6 to about 5 whereas the pH increased to about 7 when the plants were adequately supplied with Zn. In Zn deficient plants the pH decrease was associated with a shift in the cation-anion uptake ratio in favour of cation uptake. Of the major ions, uptake of Ca²⁺ and K⁺ was either not affected or only slightly lowered whereas NO₃ ^- uptake was drastically decreased in Zn deficient plants. Although the Zn nutritional status of plants hardly affected the NO₃ ^- concentrations in the plants, the leakage of NO₃ ^- from roots of Zn deficient plants into a diluted CaCl₂ solution was nearly 10 times higher than that of plants adequately supplied with Zn. In contrast to Zn deficiency, Mn deficiency in cotton plants neither affected NO₃ ^- uptake nor the pH of the nutrient solution.The results indicate that, probably as a consequence of the role of Zn in plasma membrane integrity and nitrogen metabolism, when Zn is deficient in dicotyledonous species net uptake of NO₃ ^- is particularly depressed which in turn results in an increase in cation-anion uptake ratio and a corresponding decrease in external pH. The ecological relevance of this rhizosphere acidification is discussed.     

The role of intracellular pH in the regulation of cation exchanges in yeast

1. When yeast oxidizes propan-2-ol in the presence of KCl no uptake of K⁺ occurs. 2. When propionate is added to suspensions containing propan-2-ol, or if the suspensions are bubbled with CO₂, a considerable uptake of K⁺ occurs. 3. Maximum K⁺ uptake occurs at a propionate concentration of 2mm. 4. The addition of 20mm-propionate to the suspension lowers the intracellular pH of the yeast from a resting value in the region of 6.2 to approx. 5.6. 5. When K⁺ uptake is measured in the presence of 20mm-propionate, progressive changes in the rate of K⁺ uptake and intracellular pH occur. The optimum rate of K⁺ uptake occurs at an intracellular pH of 5.70. 6. The effect of both intra- and extra-cellular pH on K⁺–K⁺ exchange was studied and an optimum rate was found at an extracellular pH of 5.35, the corresponding intracellular pH being 6.44. 7. When a Na⁺-loaded yeast oxidizes propan-2-ol in the presence of KCl, a steady efflux of Na⁺ and influx of K⁺ occurs. The addition of 10mm-propionate to the suspension markedly inhibited the Na⁺ efflux but only slightly decreased the K⁺ influx. 8. The effect of both extra- and intra-cellular pH on Na⁺ efflux was studied with propan-2-ol and with glucose. The results can be best interpreted in terms of intracellular pH changes, and an optimum was obtained at approx. pH6.40.     

Regulation of Nitrate Uptake in Penicillium chrysogenum by Ammonium Ion

A nitrate uptake system is induced (along with nitrate reductase) when NH₄⁺-grown Penicillium chrysogenum is incubated with inorganic nitrate in synthetic medium in the absence of NH₄⁺. Nitrate uptake and nitrate reduction are probably in steady state in fully induced mycelium, but the ratios of the two activities are not constant during the induction period. Substrate concentrations of ammonium cause a rapid decay of nitrate uptake and nitrate reductase activity. The two activities are differentially inactivated (the uptake activity being more sensitive). Glutamine and asparagine are as effective as NH₄⁺ in suppressing nitrate uptake activity. Glutamate and alanine were about half as effective as NH₄⁺. Cycloheximide interferes with the NH₄⁺-induced decay of nitrate uptake activity. The ammonium transport system is almost maximally deinhibited (or derepressed) in nitrate-grown mycelium.     

Sodium/potassium selectivity and pleiotropy in stl2, a highly salt-tolerant mutation of Ceratopteris richardii

The roles of Na⁺ and K⁺ (Rb⁺) uptake were further studied in a NaCl-tolerant strain of Ceratopteris richardii containing the stl2 mutation by direct comparison with the wild-type strain. In addition to Na⁺ tolerance, stl2 also confers tolerance to Mg²⁺ and sensitivity to K⁺. In addition to higher K⁺ (Rb⁺) uptake at concentrations commonly associated with low-affinity K⁺ transport, stl2 maintained higher uptake down to 0·1 mol m^–3 Rb⁺. Up to a 25-fold excess of Na⁺ had little effect in either genotype on K⁺ (Rb⁺) uptake at low concentrations, i.e. 0·2 and 0·5 mol m^–3 RbCl. Pretreatment with K⁺ (20 mol m^–3) inhibited uptake of K⁺ (Rb⁺) in the wild type, whereas concurrent inclusion of K⁺ inhibited uptake of Rb⁺ more in stl2. In the absence of K⁺, Na⁺ uptake (0·01–60 mol m^–3) was nearly identical in the wild type and stl2. K⁺ inhibited Na⁺ uptake more effectively in stl2 than the wild type, especially at 60 mol m^–3 Na⁺. Greater inhibition of K⁺ uptake in stl2 occurred with MgCl₂ or TEA (tetraethylammonium chloride) preincubation or with simultaneous inclusion of Al³⁺ (Al₂SO₄). The higher effective velocity of K⁺ uptake at a wide range of concentrations and the enhanced selectivity for K⁺ and against Na⁺ contribute to the preservation of higher cytosolic K⁺ and lower Na⁺ under salinity stress.     

The uptake and metabolism of methylamine by N2-fixing cyanobacteria

The cyanobacteria Anabaena variabilis and Nostoc CAN showed a biphasic pattern of ¹⁴CH₃NH ₃ ⁺ uptake at external pH values of 7.0 and 9.0. The initial phase of uptake, which was independent of metabolism of ¹⁴CH₃NH ₃ ⁺ , was attributed to uptake via a CH₃NH ₃ ⁺ (NH ₄ ⁺ ) transport system at pH 7.0 and probably to passive diffusion of uncharged CH₃NH₂ and trapping by protonation at pH 9.0. The second slower phase of uptake was attributed to metabolism of CH₃NH ₃ ⁺ via glutamine synthetase to form -methylglutamine which accumulates. Anabaena cylindrica showed an initial rapid uptake at pH 7.0 and pH 9.0 but metabolism of ¹⁴CH₃NH ₃ ⁺ was undetectable at pH 7.0 and was barely detectable at pH 9.0. Pretreatment of A. variabilis with l-methionine-d,l-sulphoximine to inactivate glutamine synthetase, inhibited the second phase of ¹⁴CH₃NH ₃ ⁺ uptake at both pH 7.0 and pH 9.0 and the accumulation of -methylglutamine but had no effect on the first phase of uptake. Following transfer of A. variabilis to darkness the initial phase of ¹⁴CH₃NH ₃ ⁺ uptake at pH 7.0 and 9.0 was unaffected but the subsequent metabolism via glutamine synthetase was inhibited.Abbreviations MSX l-methionine-d,l-sulphoximine - GS glutamine synthetase     

Competitive Al Inhibition of Net Mg Uptake by Intact Lolium multiflorum Roots : II. Plant Age Effects

Rhizotoxicity of Al is more pronounced in younger plants. Effects of Al on nutrient uptake by plants of different age are poorly understood. The depletion technique was used to monitor net Mg²⁺ uptake from nutrient solutions by intact 15- and 35-day-old plants of two ryegrass (Lolium multiflorum Lam.) cultivars. Lowering the pH from 6.0 to 4.2 decreased the maximum net ion influx without affecting K_m. Aluminum at 6.6 micromolar Al³⁺ activity increased K_m indicating competitive inhibition. The effects of pH and 6.6 micromolar Al³⁺ on net Mg²⁺ uptake were much larger in 15- than in 35-day-old plants. Aluminum at 26 micromolar Al³⁺ activity competitively inhibited net Mg²⁺ uptake by 35-day-old plants, while causing time- and external Mg²⁺ activity-dependent net Mg²⁺ efflux from 15-day-old plants. The equilibrium constant (K_i) of a reversible combination of postulated plasmalemma Mg²⁺ transporter and Al³⁺ was calculated to be 2 and 5 micromolar Al³⁺ activity for 15-day-old plants of Wilo and Gulf ryegrass, respectively, and 21 micromolar Al³⁺ activity for 35-day-old plants of both cultivars. The Al³⁺-mediated increase in K_m was larger for 15-day-old plants of the Al-sensitive cultivar `Wilo' than of the more Al-tolerant cultivar `Gulf,' while Al³⁺ affected 35-day-old plants of both cultivars to the same extent.     

Methylammonium uptake by Escherichia coli: evidence for a bacterial NH4+ transport system.

Energy-dependent concentrative uptake of ¹⁴CH₃NH₃⁺ by cells of $\text{Escherichia coli}$ provides preliminary evidence for one or more transport systems for NH₄⁺ uptake. NH₄⁺, but not glutamic acid, inhibited the uptake of ¹⁴CH₃NH₃⁺. Varying the pH for the uptake assays exposed two apparent systems: one maximally functioning at pH 7 that was strongly inhibited by cyanide or by the uncoupler $\text{m}$-chlorophenyl carbonylcyanide hydrazone and another maximally functioning at pH 9 and resistant to cyanide or $\text{m}$-chlorophenyl carbonylcyanide hydrazone. Kinetic analysis showed considerable experimental variability from day to day. Often simple Michaelis-Menten kinetics were not followed, but NH₄⁺ was reproducibly a stronger inhibitor of uptake of ¹⁴CH₃NH₃⁺ than was nonradioactive CH₃NH₃⁺.     

Effects of sodium on mineral nutrition in rose plants

The effects of sodium (Na⁺) ion concentration on shoot elongation, uptake of ammonium (NH₄⁺) and nitrate (NO₃^?) and the activities of nitrate reductase (NR) and glutamine synthetase (GS) were studied in rose plants (Rosa hybrida cv. “Lambada”). The results showed that shoot elongation was negatively correlated with sodium concentration, although no external symptoms of toxicity were observed. Nitrate uptake decreased at high sodium levels, specifically at 30 meq litre4 of sodium. As flower development was normal under high saline conditions, this could suggest that nitrogen was being mobilised from shoot and leaf reserves. Ammonium uptake was not affected by any of the salt treatments applied probably because it diffuses through the cell membrane at low concentrations. Nitrate reductase activity was reduced by 50% at 30 meq litre 1 compared with control treatment, probably due to a decrease in the free nitrate related to nitrate uptake pattern. None of the salt treatments used affected total leaf GS activity (both chloroplastic and cytosolic isoforms) or leaf NPK mineral contents. Nitrate reductase activity in leaves increased at 10 meq litre^?1 of sodium and GS activity in roots (cytosolic isoform only) followed the same pattern as NR. It is suggested that the activation of both enzymes at low salt level could be attributed to the beneficial effect of increased sulphur in the nutrient solutions.