FURTHER ASPECTS OF THE RELATIONSHIP BETWEEN NICKEL TOXICITY AND IRON SUPPLY

Absorption of nickel by oat plants increased with increasing pH for a fixed iron supply. Nickel uptake and toxicity symptoms (necrosis and chlorosis) were both reduced when the concentration of iron in the nutrient solution was high. Nickel-iron ratio in the nutrient solution. For solutions with the same nickel-iron ratio, toxicity symptoms increased with increase in the absolute amount of nickel. There was a linear relationship between the degree of necrotic symptoms and the nickel-iron ratio in the plant.
Nickel consistently reduced the iron content of roots and tops. In the absence of nickel, the iron content of the roots but not of the tops, increased with iron supply. In nickel-toxic plants, the magnesium, calcium and phosphorus contents of the tops and the potassium, calcium and phosphorus contents of the roots were higher than in healthy plants, but the potassium content of the tops and the magnesium content of the roots were lower.
Similar results were found with tomato.     

TRACE-ELEMENT TOXICITIES IN OAT PLANTS

Excessive amounts of nickel, cobalt, chromium, copper, zinc, manganese, molybdenum and aluminium in nutrient solutions supplied to oat plants in sand culture produce ( a ) chlorosis and ( b ) other symptoms specific to the element involved. The specific symptoms are distinct for each metal, although those of cobalt and nickel might be confused.
The toxic effects of nickel, cobalt, copper, zinc, manganese and molybdenum are associated with high concentrations of the element in the leaf tissue, but this is not always so with chromium and aluminium.
The toxic effects of nickel, chromium, copper and molybdenum are associated with a reduced nitrogen content of the plant. Nickel, cobalt, chromium, zinc and manganese increase the concentration of phosphorus in the tissue whilst aluminium decreases it, probably to a deficiency level.
Aluminium reduces the intensity of toxic symptoms produced by nickel—probably by reducing the uptake of nickel and phosphorus. Copper effectively reduces the leaf necrosis produced by nickel, but not the nickel content of the leaf tissue; it is suggested that one factor in nickel toxicity may be inhibition of one or more functions of copper. The other elements slightly increase chlorosis and some increase necrosis.
The order of activitjl of the elements in producing chlorosis is found to be Ni>Cu>Co>Cr>Zn>Mo>Mn. This order, which is related to that giving yield reduction and is similar to the order of stability of metal complexes, is discussed in relation to induced iron deficiency.     

Effect of iron chlorosis-inducing factors on the pH of the cytoplasm of sunflower (Helianthus annuus)

Summary Growth chamber experiments with sunflower in nutrient solution were performed to investigate the effect of phosphorus and bicarbonate in inducing iron chlorosis.Iron chlorosis as proved by lower dry matter yield and reduced chlorophyll content was induced by bicarbonate alone and more pronounced by a combination of bicarbonate and phosphate, but not by phosphate alone.Iron content of roots and aerial plant parts was reduced by bicarbonate in all experiments, but only in one experiment by phosphate alone.Bicarbonate in the nutrient medium increased the pH of the cytoplasm in leaf cells, while phosphate had no effect.A daily adjustment of the pH in the nutrient medium to a value comparable to that in the bicarbonate trial, did not affect the pH of the cytoplasm.It is concluded that the pH of the cytoplasm plays an important role in establishing plant resistance or susceptibility to Fe chlorosis.     

Nitrate does not result in iron inactivation in the apoplast of sunflower leaves

It has been hypothesized that nitrate (NO(3)(-)) nutrition might induce iron (Fe) deficiency chlorosis by inactivation of Fe in the leaf apoplast (H.U. Kosegarten, B. Hoffmann, K. Mengel [1999] Plant Physiol 121: 1069-1079). To test this hypothesis, sunflower (Helianthus annuus L. cv Farnkasol) plants were grown in nutrient solutions supplied with various nitrogen (N) forms (NO(3)(-), NH(4)(+) and NH(4)NO(3)), with or without pH control by using pH buffers [2-(N-morpholino)ethanesulfonic acid or 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid]. It was shown that high pH in the nutrient solution restricted uptake and shoot translocation of Fe independently of N form and, therefore, induced Fe deficiency chlorosis at low Fe supply [1 micro M ferric ethylenediaminedi(O-hydroxyphenylacetic acid)]. Root NO(3)(-) supply (up to 40 mM) did not affect the relative distribution of Fe between leaf apoplast and symplast at constant low external pH of the root medium. Although perfusion of high pH-buffered solution (7.0) into the leaf apoplast restricted (59)Fe uptake rate as compared with low apoplastic solution pH (5.0 and 6.0, respectively), loading of NO(3)(-) (6 mM) showed no effect on (59)Fe uptake by the symplast of leaf cells. However, high light intensity strongly increased (59)Fe uptake, independently of apoplastic pH or of the presence of NO(3)(-) in the apoplastic solution. Finally, there are no indications in the present study that NO(3)(-) supply to roots results in the postulated inactivation of Fe in the leaf apoplast. It is concluded that NO(3)(-) nutrition results in Fe deficiency chlorosis exclusively by inhibited Fe acquisition by roots due to high pH at the root surface.     

THE RELATIONSHIP BETWEEN NICKEL-TOXICITY SYMPTOMS AND THE ABSORPTION OF IRON AND NICKEL

During a 70-day experimental period from germination to maturity, the iron content of oat plants that showed symptoms of nickel toxicity changed little, but the nickel content increased rapidly for about 30 days and then decreased slowly. Necrosis varied little with time, while chlorosis increased in severity for 40 days, then decreased until unfolding young leaves were no longer chlorotic. This change in chlorotic symptoms was correlated with the nickel-iron ratio in the plant.
Autoradiographs of leaves from plants supplied with radioactive iron showed that necrotic areas in the leaf matched areas in the autoradiograph having a very low content. Chlorotic areas were found to correspond with areas whose iron content was lower than that of healthy tissue. More iron was found in the veins than in the interveinal tissue, and its distribution was the same whether supplied as ferric citrate or in a chelated form.
The concentration of iron in mature leaves from oat plants growing in a nickel toxic soil was lowest in the necrotic areas of the leaf, suggesting a migration of nutrients out of this dying tissue.     

Iron deficiency response in relation to iron uptake in two cultivars and a local strain ofMentha arvensisL.

The tolerance to iron-deficiency stress and the iron uptake were studied in two Japanese mint(Mentha arvensis L.) cultivars MAS1 and MS77 and their local strain MA2. A considerable reduction of pH of the nutrient medium in MAS1 and MS77 treatments associated with different degree of chlorosis was found. A rapid recovery from chlorosis was found only in MS77 and to some degree in MAS1, but not in the MA2. The results indicated that iron uptake and translocation were inversely related to iron stress tolerance.     

Mechanism of Fe uptake by the leaf symplast: Is Fe inactivation in leaf a cause of Fe deficiency chlorosis?

The mechanism of iron (Fe) uptake from the leaf apoplast into leaf mesophyll cells was studied to evaluate the putative Fe inactivation as a possible cause of Fe deficiency chlorosis. For this purpose, sunflower (Helianthus annuus L.) and faba bean plants (Vicia faba L.) were precultured with varied Fe and bicarbonate (HCO ₃ ^- ) supply in nutrient solution. After 2–3 weeks preculture, Fe^III reduction and ⁵⁹Fe uptake by leaf discs were measured in solutions with Fe supplied as citrate or synthetic chelates in darkness. The data clearly indicate that Fe^III reduction is a prerequisite for Fe uptake into leaf cells and that the Fe nutritional status of plants does not affect either process. In addition, varied supply of Fe and HCO ₃ ^- to the root medium during preculture had no effect on pH of the xylem sap and leaf apoplastic fluid. A varied pH of the incubation solution had no significant effect on Fe^III reduction and Fe uptake by leaf discs in the physiologically relevant pH range of 5.0–6.0 as measured in the apoplastic leaf fluid. It is concluded that Fe inactivation in the leaf apoplast is not a primary cause of Fe deficiency chlorosis induced by bicarbonate. This revised version was published online in June 2006 with corrections to the Cover Date.     

Varietal differences in mungbean (Vigna radiata) for growth, yield, toxicity symptoms and cadmium accumulation

Increased cadmium (Cd) contamination of soils resulting from industrial activities is critical to crop production. The objective of this study was to find varietal differences for foliar chlorosis and necrosis, growth and Cd accumulation in mungbean (Vigna radiata). Despite substantial varietal differences, increased Cd levels reduced the shoot and root dry weight and the number and area of leaves at different growth stages. Applied Cd stress produced the foliar symptoms such as marginal and intervein chlorosis and scattered necrotic spots on younger leaves while accelerating the senescence of older leaves. Slope of regression equation and correlations of shoot Cd content with foliar Cd toxicity revealed that leaf chlorosis was more damaging than necrosis. At maturity, number of pods per plant and seeds per pod were maximally reduced to 37% and 26%, while 100‐seed weight, seed yield and harvest index showed 61%, 79% and 54% reduction, respectively, as a result of Cd toxicity. Results suggested that although varietal difference exists, the accumulated Cd is mainly toxic to the mesophyll tissue, most probably by interfering with the uptake of essential nutrients, thereby reducing growth and yield at various stages. Therefore, selection programmes based on foliar toxicity criteria may be beneficial for better utilisation of Cd‐polluted soils.     

INVESTIGATIONS REGARDING THE NATURE OF THE INTERACTION BETWEEN IRON AND MOLYBDENUM OR VANADIUM IN NUTRIENT SOLUTIONS WITH AND WITHOUT A GROWING PLANT

Iron offset the toxicity of molybdenum or vanadium in nutrient solutions more effectively when it was supplied at the same time as the molybdenum or vanadium than when it was given separately in alternate 3-day periods.
Allowing nutrient solutions of pH 4.6 containing high concentrations of iron, with or without vanadium, to stand for 4 days before use did not delay the restoration of colour to chlorotic plants, but even z days' standing reduced the iron content of their roots and the vanadium content of both shoot and root. The presence of vanadium had little effect on iron uptake.
In parallel experiments with molybdenum, standing the solutions for 7–9 days before use delayed colour recovery, but shorter periods had no effect. Standing for z days or longer greatly reduced the iron content of the root, but the molybdenum content was unaffected or increased. High molybdenum greatly increased the iron in the root, but had little effect on that in the shoot.
Precipitation of iron in the nutrient solution was delayed by high concentrations of either ammonium or sodium molybdate if the initial pH was 4.6, but not if it was 6.6. Vanadium had no influence on the precipitation of iron at pH 4.6.
At least part of the compensating action of iron on molybdenum or vanadium toxicity would appear to take place outside the plant.     

A genetically related response to iron deficiency stress in muskmelon

A mutant muskmelon (Cucumis melo L.) with characteristic Fe-deficiency chlorosis symptoms was compared to related cultivars in its ability to obtain Fe via the widely known Fe-stress response mechanisms of dicotyledonous plants. The three cultivars (fefe, the Fe-inefficient mutant; Mainstream and Edisto, both Fe efficient plants) were grown in nutrient solution in either 0 or 3.5 mg L^-1 Fe as FeCl₃. None of the three cultivars released reductants or phytosiderophores, but both Edisto and Mainstream produced massive amounts of H+ ions to reduce and maintain the pH of nutrient solutions below pH 4.0. The roots of these two Fe-efficient cultivars were also capable of reducing Fe³⁺ to Fe²⁺. These responses maintained green plants, resulted in high leaf Fe in both Edisto and Mainstream, and produced Mn toxicity in Mainstream. The lack of Fe-deficiency stress response in fefe not only affected leaf Fe concentration and chlorosis, but also resulted in reduced uptake of Mn. The importance of reduced Fe (Fe²⁺) to the Fe-efficient cultivars was confirmed by growing the cultivars with BPDS (4, 7-diphenyl-1, 10-phenanthroline disulfonic acid, a ferrous chelator) and EDDHA [ethylene-diamine di (0-hydroxphenylacetic acid)] (a ferric chelator), and observing increased chlorosis and reduced Fe uptake in BPDS grown plants. The Fe-deficiency response observed in these cultivars points out the diversity of responses to Fe deficiency stress in plants. The fefe mutant has a limited ability to absorb Fe and Mn and perhaps could be used to better understand Mn uptake in plants.     

Factors effecting the toxicity of nitrite nitrogen to tomatoes

Experiments were done to study the effects of nitrite nitrogen on nutrient absorption and organic acid content of tomatoes (Keystone) grown in sand culture. The effects of root aeration, magnesium and iron supply on the symptoms of nitrite toxicity were also studied. Nutrient solutions were standardised to pH 4.5 and contained from 0–250 ppm nitrite nitrogen. Increasing the concentration of nitrite nitrogen decreased dry matter yields, total acidity, the concentration of nitrogen, phosphorus and potassium in tomato plants, and increased the chlorosis of leaves and the lignification of roots. Shortage of iron, magnesium, and poor root aeration caused toxicity symptoms to appear at a smaller concentration of nitrite nitrogen and increased the severity of the symptoms.     

Relationship between iron chlorosis and alkalinity in Zea mays

Mengel, K. and Geurtzen, G. 1988. Relationship between iron chlorosis and alkalinity in Zea mays . - Physiol. Plant. 72: 460–465.
Maize ( Zea mays L. cv. Anjou 21) grown in nutrient solution with Fe-EDTA and with nitrate as the sole nitrogen source showed typical Fe-chlorosis symptoms after a growth period of 14–21 days. Alkalinity in roots, stems and leaves of the chlorotic plants was high. Transferring the chlorotic plants from the nitrate-containing nutrient solution to a solution of (NH₄)₂SO₄ resulted in a regreening of leaves within 2–3 days which was associated with a decrease in solution pH, a decrease in alkalinity of plant parts, a translocation of Fe from roots to tops and a release of Fe into the outer solution. Similar effects were obtained when Fe chlorotic plants were transferred to a dilute HO solution with pH 3.5.
Spraying chlorotic leaves with indoleacetic acid or with fusicoccin led also to a regreening of leaves without having a major effect on leaf alkalinity.
Interpretation of the experimental results is based on the assumption that nitrate as sole N source leads to a high pH level in the apoplast resulting in the precipitation of Fe compounds, probably Fe oxide hydrate. Ammonium nutrition has the reverse effect since it lowers the apoplast pH and this can result in the dissolution of Fe compounds. Application of indoleacetic acid as well as fusicoccin supposedly stimulates the proton pumps in the plasmalemma of the leaf tissue. The resulting decrease in apoplast leaf pH in the microenvironment also leads to a dissolution of Fe compounds in the apoplast and thus promotes the uptake of Fe by the symplasm.     

Effects of the Environment on the Symptom Pattern of Nickel Toxicity in the Oat Plant

Under conditions of nickel toxicity in oats (Avena byzantina),a predisposing condition for the development of chlorosis isinduced in areas of the leaf before emergence from the coleoptileor the enclosing leaf sheath. These areas give rise to chloroticbands which develop on the emerged leaf a little over 24 h afteremergence of the tissue. Alternating light and dark are essentialfor the development of chlorotic bands. The evidence indicatesthat the potentially green tissue is that which is developingunder the coleoptile during the day and emerges later in theday or early in the night, while the potentially chlorotic tissueis that which develops under the coleoptile during the nightand emerges from the top of the coleoptile later in the nightor during the following daylight hours before midday. Plants containing high levels of nickel contain higher levelsof protochlorophyll and lower levels of of chlorophyll thancontrol plants. The visual symptom of nickel toxicity is influenced by the lengthof the light and dark periods. The nature of these effects isdiscussed. Avena byzantina, oat, nickel toxicity symptoms, chlorosis     

Does high bicarbonate supply to roots change availability of iron in the leaf apoplast?

The role of the leaf apoplast in iron (Fe) uptake into the leaf symplast is insufficiently understood, particularly in relation to the supposed inactivation of Fe in leaves caused by elevated bicarbonate in calcareous soils. It has been supposed that high bicarbonate supply to roots increases the pH of the leaf apoplast which decreases the physiological availability of Fe in leaf tissues. The study reported here has been carried out with sunflower plants grown in nutrient solution and with grapevine plants grown on calcareous soil under field conditions. The data obtained clearly show that the pH of the leaf apoplastic fluid was not affected by high bicarbonate supply in the root medium (nutrient solution and field experiments). The concentrations of total, symplastic and apoplastic Fe were decreased in chlorotic leaves of both sunflower (nutrient solution experiment) and grapevine plants in which leaf expansion was slightly inhibited (field experiment). However, in grapevine showing severe inhibition of leaf growth, total Fe concentration in chlorotic leaves was the same or even higher than in green ones, indicative to the so-called `chlorosis paradox'. The findings do not support the hypothesis of Fe inactivation in the leaf apoplast as the cause of Fe deficiency chlorosis since no increase was found in the relative amount of apoplastic Fe (% of total leaf Fe) either in the leaves of sunflower or grapevine plants. It is concluded that high bicarbonate concentration in the soil solution does not decrease Fe availability in the leaf apoplast.     

THE INFLUENCE OF THE pH OF THE NUTRIENT SOLUTION AND THE FORM OF IRON SUPPLY ON THE COUNTERACTION OF IRON DEFICIENCY IN PEAS, SOYBEAN AND FLAX BY HIGH CONCENTRATIONS OF MOLYBDENUM

The prevention of chlorosis in flax by high concentrations of molybdenum in a nutrient solution was associated with a delay in the precipitation of iron from ferric citrate, a slower drift of pH towards alkalinity and an increase in the iron content of the root. These effects were greater with ammonium than with sodium molybdate and occurred with solutions started at pH 4.6 but not at pH 6.6.
When FeEDTA was the source of iron, a similar delay in pH drift in the solution and accumulation of iron in the root occurred, but there was no chlorosis or precipitation of iron in the control treatment, so the effect of high molybdenum could not be fully determined.
When ferric chloride was used, high molybdenum did not prevent chlorosis nor delay iron precipitation or cause accumulation of iron in the root, though the rate of pH drift resembled that of solutions containing the organic forms of iron.
Similar results were obtained with peas and soybeans receiving high molybdenum treatment, but suppression of chlorosis was only temporary.
It is suggested that the capacity of molybdenum to offset chlorosis is due to the formation, in acid solution, of a complex with phosphorus which renders iron more available by delaying the formation of ferric phosphate. This seems to occur only when iron is supplied in the organic form.     

Iron availability in plant tissues-iron chlorosis on calcareous soils

The article describes factors and processes which lead to Fe chlorosis (lime chlorosis) in plants grown on calcareous soils. Such soils may contain high HCO₃ ^- concentrations in their soil solution, they are characterized by a high pH, and they rather tend to accumulate nitrate than ammonium because due to the high pH level ammonium nitrogen is rapidly nitrified and/or even may escape in form of volatile NH₃. Hence in these soils plant roots may be exposed to high nitrate and high bicarbonate concentrations. Both anion species are involved in the induction of Fe chlorosis.Physiological processes involved in Fe chlorosis occur in the roots and in the leaves. Even on calcareous soils and even in plants with chlorosis the Fe concentration in the roots is several times higher than the Fe concentration in the leaves. This shows that the Fe availability in the soil is not the critical process leading to chlorosis but rather the Fe uptake from the root apoplast into the cytosol of root cells. This situation applies to dicots as well as to monocots. Iron transport across the plasmamembrane is initiated by Fe^III reduction brought about by a plasmalemma located Fe^III reductase. Its activity is pH dependent and at alkaline pH supposed to be much depressed. Bicarbonate present in the root apoplast will neutralize the protons pumped out of the cytosol and together with nitrate which is taken up by a H⁺/nitrate cotransport high pH levels are provided which hamper or even block the Fe^III reduction.Frequently chlorotic leaves have higher Fe concentrations than green ones which phenomenon shows that chlorosis on calcareous soils is not only related to Fe uptake by roots and Fe translocation from the roots to the upper plant parts but also dependent on the efficiency of Fe in the leaves. It is hypothesized that also in the leaves Fe^III reduction and Fe uptake from the apoplast into the cytosol is affected by nitrate and bicarbonate in an analogous way as this is the case in the roots. This assumption was confirmed by the highly significant negative correlation between the leaf apoplast pH and the degree of iron chlorosis measured as leaf chlorophyll concentration. Depressing leaf apoplast pH by simply spraying chlorotic leaves with an acid led to a regreening of the leaves.     

Xylary pH and Reduction Potential Levels of Iron-stressed Silver Maple (Acer saccharinum L.)

Xylary fluid pH and reduction potentials were measured on silver maple (Acer saccharinum L.) grown under Fe and pH stress. Although pH and reduction potential (millivolt/59.2) varied significantly in the nutrient solution, xylary pH and reduction potential remained constant. It was concluded that changes in the pH and reduction potential in the xylary fluid of silver maple are not responsible for iron chlorosis.     

Zinc, Iron, and Chlorophyll Metabolism in Zinc-toxic Corn

Zinc toxicity and Zn-Fe interactions were studied in corn (Zea mays L. var. Barbecue hybrid) grown in hydroponic culture. High Zn greatly reduced the root and shoot fresh weights; increasing Fe largely, but not completely, restored normal growth. Correlation analyses of root and leaf Zn and Fe contents suggested that Zn may interfere with the translocation of Fe; however, Zn toxicity was not associated with a diminished leaf Fe content. Fe did appear to retard both the absorption and the translocation of Zn. The chlorosis of Zn-toxic plants is not attributable to diminshed total leaf Fe; however, this chlorosis is relieved by increasing nutrient Fe. Zn and Fe probably do interact at some site.     

A physiological study ofTeucrium scorodonia ecotypes which differ in their susceptibility to lime-induced chlorosis and iron-deficiency chlorosis

Summary Edaphic ecotypes ofTeucrium scorodonia have been shown which differ in their susceptibility to lime-induced chlorosis. Plants especially resistant or susceptible to lime-induced chlorosis were found to be similarly resistant or susceptible to iron-deficiency chlorosis. Differences were found in the chlorophyll-iron and dry weight-chlorophyll relationships of the leaves of green and chloroticTeucrium plants, similar effects being produced by growth on a calcareous soil, in iron-deficient culture or by bicarbonate treatment. Chlorotic leaves had less chlorophyll per unit iron but had a greater dry weight per unit chlorophyll than green material. Chlorotic leaves were found to be reduced in both leaf area and dry weight compared with green ones, the reduction in dry weight being the greatest. Common root abnormalities were noted in chlorotic material induced by the above three methods.Evidence was produced which suggested that the difference between chlorosis-resistant and susceptible plants lay in qualitative differences in their iron transport compounds produced within the rootstock. Differential iron uptake was not suggested as a cause of the differences in behaviour. There was, however, evidence of a key role of the root iron pool in population differences in chlorosis susceptibility.Bicarbonate was found to suppress first iron uptake and then iron translocation. A possible causal role of the bicarbonate ion in lime-induced chlorosis was suggested through these effects and through its possible effect on the production of iron-transport compounds.     

PHOTOSYNTHETIC AND CHLOROPLAST ULTRASTRUCTURAL CONSEQUENCES OF MANGANESE DEFICIENCY IN SOYBEAN

Soybean plants (Glycine max ‘Harasoy’) grown in horticultural perlite with a manganese-deficient nutrient solution, developed typical deficiency symptoms of interveinal chlorosis and necrosis. A decrease in photosynthetic rate, dry leaf weight, leaf area, chlorophyll content, and chloroplast number was observed in plants displaying the deficiency. Transmission electron microscopy of manganese-deficient leaf tissue revealed disorganization in the chloroplast lamellar network, nonhomogeneous distribution of the stroma, and a reduction in quantities of starch, which became increasingly more acute from nodes three to five in plants 28 days postgermination. The data suggest that manganese functions both as a requirement in the photosynthetic apparatus and as a structural component in the lamellar membrane.