THE RELATIONSHIP BETWEEN NICKEL TOXICITY AND IRON SUPPLY

The influence of varying levels of iron and substrate pH on the uptake of nickel and the intensity of toxicity symptoms in oat plants have been investigated using sand-and water-culture techniques.
Nickel-toxicity symptoms (both necrosis and chlorosis) are less severe when the concentration of iron in the nutrient solution is high. The reduction in degree of necrosis is related to a reduced content of nickel in the leaf blades, whilst that of chlorosis is related to the Ni/Fe ratio in the leaf blades—an internal antagonism being indicated in the latter case.
A reciprocal relationship exists between the nickel and iron contents of the leaf blades; the nickel content is materially reduced by high concentrations of iron in the nutrient solution, and the iron content by nickel, the former being the more pronounced effect.
Uptake of nickel increases with increasing pH for a constant iron level in the substrate, although the degree of necrotic symptoms is similar over pH range 4–7. Iron uptake is reduced by both nickel and increasing pH and results in chlorosis at pH values of 5·5 and above.
For a constant level of iron supply the phosphate content of the stem extracts is higher the greater the degree of nickel toxicity; the phosphorus status of the plant may be a factor in producing nickel toxicity but if so, it has to be considered in relation to other factors.     

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.     

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     

STUDIES ON THE UPTAKE AND TRANSLOCATION OF STREPTOMYCIN BY PEACH SEEDLINGS

Measurements of the antibiotic activity in the foliage of peach seedlings, kept with their roots in mineral nutrient solutions containing streptomycin, showed that streptomycin passed into the foliage and sometimes reached concentrations above those of the solutions surrounding the roots. The amount of the antibiotic in leaves varied with the time that roots were kept in the solutions and with the streptomycin concentrations of the solutions. Lower leaves contained much more than upper leaves.
Adding macerated leaf tissue to streptomycin solutions decreased the amount of streptomycin detectable in the supernatant liquid more than could be accounted for by dilution or alteration of pH.
Streptomycin first affected plants by decreasing apical growth, but at higher concentrations it caused chlorotic and necrotic areas on leaves and stems.     

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.     

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.     

Selenium delays leaf senescence in oilseed rape plants

Effect of selenium on leaf senescence was studied in oilseed rape plants treated with 10 μM Na₂SeO₄ at a rosette growth stage. In addition to developmental senescence, N deficiency and leaf detachment were used for induction of senescence. Nonphotochemical quenching declined in old leaves as senescence became more advancing but rose progressively in the plants supplied by Se. The total carbohydrate and protein pools decreased with leaf age, while increased by the Se treatment. However, during senescence induced by N deficiency, Se did not change remarkably the C and N metabolism, but delayed senescence mainly through protection of plants from photoinhibitory effects. After detachment, untreated leaves became chlorotic and necrotic, while the Se-treated ones remained fairly green. Our results demonstrated that Se delayed leaf senescence by a maintaining or even improving photochemical activities. During developmental senescence, the Se effect on the extending life span of the leaves was additionally linked to the metabolic regulation of senescence.     

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.     

Wheat Resistance to Bacterial Leaf Streak Mediated by Silicon

Leaf streak, caused by Xanthomonas translucens pv. undulosa, is the major bacterial disease of wheat in Brazil and other countries worldwide. This study aimed to evaluate the effect of silicon (Si) on disease development and the biochemical mechanisms possibly involved in resistance potentialized by this element. Plants of cv. BR‐18, susceptible to leaf streak, were grown in plastic pots containing Si‐deficient soil amended with either calcium silicate (+Si) or calcium carbonate (?Si). The content of Si increased (P ≤ 0.05) by 96.5% for the +Si when compared with ?Si treatment. There was no difference (P ≥ 0.05) between Si treatments for calcium content on leaf tissue, so variations in Si accounted for differences in the level of resistance to leaf streak. There was no difference (P ≥ 0.05) between Si treatments for incubation period, latent period, necrotic leaf area, and severity estimated by the software quant . However, chlorotic leaf area was reduced (P ≤ 0.05) by 50.2% for the +Si when compared with ?Si treatment. There was no difference (P ≥ 0.05) between Si treatments for the bacteria population on leaf tissue; however, the values seemed to be somewhat lower in the +Si treatment from 4 to 8 days after inoculation (d.a.i.) on leaves from plants supplied with Si. There was no difference (P ≥ 0.05) between Si treatments for electrolyte leakage. The concentration of total soluble phenolics and lignin‐thioglycolic acid (LTGA) derivatives did not show any apparent signs of increase during the course of infection and seemed to be slightly higher on plants not supplied with Si at the most advanced stages of bacterial infection. Chitinase activity was high at the most advanced stages of bacterial infection on leaves from +Si treatment and probably affected bacterial growth on leaf tissue. Peroxidase activity following bacterial infection was not increased by Si, but can be linked with the highest concentration of LTGA derivatives at 12 d.a.i. of plants supplied with Si. Polyphenoloxidase activity did not affect wheat resistance to leaf streak regarding of the Si treatments. The results clearly suggest that supplying Si to wheat plants can increase resistance to leaf streak possibly through an increase in tissue lignification and the participation of chitinases and peroxidases.     

VIRUSES CAUSING ROSETTE AND OTHER DISEASES IN GROUNDNUTS

From plants with a form of groundnut rosette disease, characterized by discrete areas of green and chlorotic tissue on the leaflets and here designated 'mosaic rosette', a virus was separated that produced only a mild mottle or sometimes a mottle with rare chlorotic flecks. It was separated by leaf grafts, by mechanical inoculation and by Aphis craccivora .
Plants inoculated simultaneously with the mottle virus and normal rosette virus usually developed the mosaic-rosette symptoms. When the mottle virus was introduced 14–35 days before the rosette virus, the plants failed to develop the severe chlorosis of rosette; the mottle virus thus protected the plant from rosette, and this was true whether the rosette virus was inoculated by aphids or by grafting.
Plants showing two other forms of mild mottle were collected in the field; viruses from them were readily transmitted by grafting or by mechanical inoculation, but not by A. craccivara . In plant-protection tests with one of these, it failed to protect plants from developing chlorotic symptoms when later inoculated with the rosette virus, although a form of interaction was evident and the doubly-infected plant was less severely chlorotic and less stunted than one infected with the rosette virus alone.     

Growth of Pseudomonas phaseolicola in susceptible and in resistant bean plants

Race 1 of Pseudomonas phaseolicola introduced into leaves of susceptible Canadian Wonder bean plants multiplied logarithmically for 3–5 days, reaching final populations about 10⁵–10⁶ times the original. In resistant Red Mexican, Race 1 multiplied less rapidly to give final populations about 10²–10³ times the original. Race 2 behaved in susceptible Red Mexican as did Race 1 in Canadian Wonder. Macroscopic symptoms appeared in leaves when bacterial numbers reached their maxima. When introduced into the cotyledonary node Race 1 moved more rapidly upwards than downwards, and more rapidly and farther in Canadian Wonder than in Red Mexican. But even in Canadian Wonder the bacterium appeared only sporadically above the node of the first compound leaf. It could be isolated only rarely from chlorotic haloes around necrotic areas in leaves, or from chlorotic leaves not carrying lesions. Fewer lesions developed and the bacteria multiplied less in older than in younger leaves. Addition of glucose and casein hydrolysate to inocula of Race 1, separately or together, had little effect on growth in Canadian Wonder or Red Mexican, and the bacterium grew equally well in extracts of susceptible and of resistant plants. Preinoculation of leaves with an avirulent race reduced the number of lesions caused by a virulent race inoculated later, and also reduced growth of this race in leaves of a susceptible variety.     

Some problems associated with determining iron in plants

Summary 1. Iron which is shown to be present as a contaminant on leaf surfaces of tomato can be removed completely by washing the leaves for 30 sec in 0.3% teepol and water followed by a 30 sec rinsing in distilled water.2. By using radioactive iron it is shown that there was no loss of the metal from within the plants during leaf washing. Agreement between chemical and radioassay data for iron showed that the metal was completely removed by the wash procedure.3. In tomato grown in the glasshouse consistent differences between the iron contents of normal and chlorotic leaves were obtained after leaf washing. When the plants were grown in a dustproof chamber differences in iron status of green and chlorotic leaves were obtained without washing.4. Added radioactive and carrier iron was fully recovered from acid digests of a variety of plant material, specially chosen to represent a range of silica contents.     

Diagnosis of iron deficiency in groundnut,Arachis hypogaea L.

Summary Investigations into iron deficiency have been hindered by the lack of a satisfactory diagnostic tissue test, which in turn results from the total iron content of plant tissue commonly being an unreliable index of the iron status. Our measurements of chlorotic and normal leaves of field grown groundnut (Arachis hypogaea L.) showed that total iron was unsatisfactory as the measure of iron status of plant tissue. It was found that iron status was better assessed from an estimate of the ferrous iron content of fresh leaf materials obtained by extraction with o-phenanthroline. Extractable iron content increased with leaf age. Chlorotic buds or the first fully opened leaf always contained less than 6μg extractable-Fe/g fresh tissue. Approved for publication as ICRISAT Journal Article No. 307.     

Water Stress in Leaves of Pbaseolus vulgaris Infected with Xanthomonas campestris pv. phaseoli

Infection of bean leaves ( Phaseolus vulgaris ) by Xanthomonas campestris pv. phaseoli in the field frequently resulted in the appearance of isolated flaccid areas in green leaf tissue adjacent to necrotic and chlorotic lesions. The flaccid leaf areas had significantly higher stomatal resistances compared to nearby turgid areas on the same leaf, and the turgid areas had stomatal resistances that were the same or only moderately elevated compared to those of healthy leaves. The flaccid tissues also had significantly lower relative water contents than turgid tissues on the same leaf demonstrating that pathogeninduced water stress was localized. The levels of free proline, another indicator of water stress, were directly correlated (r²= 0.556) with disease severity. The change in free proline content implied that water stress increased in direct proportion with the amount of tissue infected. Water stress may be due to the disruption of xylem elements by the invasion of X. c. phaseoli from nearby lesions. One result of xylem invasion could have been severe water deficits which were sufficient to cause stomatal closure and leaf flaccidity; however, this effect was highly localized and the remainder of the diseased leaf was either significantly less water stressed or not affected.     

Effect of Fe 3+, Zn 2+ and Mn 2+ on ferric reducing capacity and regreening process of the peach rootstock Nemaguard (Prunus persica (L.) Batsch)

The peach rootstock Nemaguard is susceptible to lime-induced iron deficiency chlorosis. Under field conditions, application of ferric chelates to the soil is effective in correcting the Fe-deficiency symptoms. The objectives of this work were to study the induction of the root ferric reducing capacity and the relationship between chlorosis and leaf Fe concentration of plants grown hydroponically under different treatments. Results showed that bicarbonate-treated plants grown with a low Fe concentration increased their reducing capacity if they received additional Fe or Zn for a short period (15 h). However, the addition of Mn had no effect. When these Mn-treated plants were changed to nutrient solution with no bicarbonate and sufficient Fe, regreening was retarded several days in relation to the other treatments. In plants grown without bicarbonate, the reducing capacity was higher in plants grown with a low amount of Fe than in plants grown with either 0 Fe or sufficient Fe. In plants grown with bicarbonate and low Fe, the leaves became chlorotic and had low Fe concentration. When these plants received higher Fe supply, the regreening of the old leaves was not complete, though they had even higher Fe concentrations that the new developing leaves which were completely green. Results are discussed in relation to the Fe or Zn requirements of plants to induce reducing capacity and the incapacity of the cells from chlorotic leaves to absorb Fe and repair metabolic and structural damages.     

THE INDUCTION OF LEAF TUMORS BY AGROBACTERIUM TUMEFACIENS

Using carborundum as an abrasive and light rubbing with a culture of Agrobacterium tumefaciens, leaves of various species of bean and tobacco develop tumors on the leaf lamina. The induction of these tumors requires wounding, the presence of a virulent strain of the bacterium and is due to the bacterium, not substances released into the bacterial culture medium during growth. Observations of the histology and cytology of these tumors on the primary leaves of pinto bean show no significant differences from the more commonly studied stem tumors. The tumors on pinto beans first appear as chlorotic nests of dividing cells which gradually accumulate chlorophyll, eventually becoming dark green in color as opposed to the surrounding leaf tissue which is completely chlorotic at this stage. Tumor development is enhanced by a dark period following inoculation while growth of the leaf is essentially stopped. The tumors thus exhibit a pattern of growth and development independent of that of the normal leaf. The number of tumors obtained on pinto bean leaves was found to depend on the concentration of bacteria in the inoculum and on the age of the plants. A sharp peak in response was observed at about 7 days from planting. Best results were obtained by adding the bacterium at the time of wounding. The tumors were shown to differ from IAA-induced leaf proliferations with respect to their point of origin on the leaf, morphology, physiology and development.     

The Association of "Pathogenesis-related" Proteins with Viral Induced Necrosis in Nicotiana sylvestris

The association of “pathogenesis-related” (PR) proteins with protection from superinfection, systemic acquired resistance and production of localized necrotic lesions was examined with a system using tobacco mosaic virus (TMV) and Nicotiana sylvestris. Leaves of N. sylvestris with a mosaic from earlier inoculation with a systemically infecting strain of TMV (TMV-C) and control plants were challenged with a necrotizing strain of TMV (TMV-P), RNA of TMV-P and turnip mosaic virus (TuMV). TMV-P virions produced localized necrotic lesions only in the dark green areas of the mosaic of TMV-C infected plants. Both RNA of TMV-P and TuMV produced localized necrotic lesions in both light green and dark green areas of the mosaic of TMV-C infected plants. All three challenge inocula produced localized necrotic lesions in previously uninoculated plants. Six days after challenge inoculation proteins were extracted from separated dark green and light green mosaic leaf tissue, and leaf material from control plants. Proteins were separated by electrophoresis in a 5 % polyacrylamide spacer gel and 10 % polyacrylamide running gel. PR proteins were found in tissue where localized necrotic lesions were produced as a result of challenge inoculation, but not in tissue that was not superinfected. PR proteins were not found in light green or dark green mosaic leaf tissue as a result of TMV-C inoculation. No PR proteins were evident in protein extracts from light green tissue challenged with TMV-P, although PR proteins were produced in dark green tissue, where necrosis occurred, from the same leaves. Systemic acquired resistance (reduction in size of lesions formed by a challenge inoculation) to TuMV or RNA of TMV-P and PR protein concentration was measured at various times in light green areas of mosaic leaves where dark green areas of the mosaic leaves had been inoculated with TMV-P. No quantitative or temporal relationship between the onset of resistance and PR protein production was found. It is concluded that PR proteins are a result of pathogen induced necrosis and not significantly involved in the mechanism(s) of viral induced resistance.     

The effect of early blight disease caused by Alternaria solani on shoot growth of young tomato plants

The effect of increasing spore concentration of Alternaria solani (Early blight disease) on the shoot growth of young tomato plants was analysed. Changes in growth were related to the severity of infection which increased with increasing inoculum. Leaf production was not affected but dry weights and especially leaf expansion were decreased. The effective leaf areas of the five inoculated leaves (L1-L5 numbered from the plant base) were drastically decreased by expanding necrotic lesions and, to a lesser extent, by premature leaf fall. Healthy leaves expanding soon after inoculation (L6, L7) were markedly affected by the disease on the lower leaves and had decreased specific leaf areas (ratio of leaf area to leaf dry weight) but later formed (from L8) leaves were less affected and had greater specific leaf areas than equivalent leaves on uninoculated plants.     

TOXIC EFFECTS OF HEAVY METALS ON CROP PLANTS GROWN IN SOIL CULTURE

A number of pot experiments were made on various crops, to study the effects on the chemical composition of the plants of treating the soil with cobalt, copper or nickel sulphates, or with the cupric or ferric salts of ethylenediamine-tetraacetic acid (EDTA).
The tops of copper sulphate-treated plants usually contained higher concentrations of copper, manganese, calcium and magnesium and lower concentrations of potassium and phosphorus, than those of control plants.
The most marked effect of cobalt on oat was to increase the concentration of Ca+Mg in the tops; nickel had a profound effect on the composition of barley tops.
Total iron concentration was reduced in oat by cobalt and copper, and in barley by copper and nickel application. Symptoms of iron deficiency appeared only in the oat plants, their incidence being correlated with the reduction in iron concentration in the tops.
The ferric salt of EDTA tended to reduce the incidence of iron deficiency symptoms produced in oat by copper sulphate, but also gave rise to distinct toxic effects. Plants given the cupric salt of EDTA contained unusually large amounts of iron.     

The effect of calcium bicarbonate on iron absorption and distribution byChrysanthemum morifolium, (Ram.)

Summary Plants grown for two weeks in high-bicarbonate nutrient solution with iron became chlorotic, absorbed less iron, and translocated a lower percentage of absorbed iron than did green plants grown under low bicarbonate with iron. Chlorotic plants, pretreated in low-bicarbonate solutions lacking iron, absorbed more iron and translocated a higher percentage to leaves than the green plants. Plants induced to chlorosis by high bicarbonate absorbed less iron after transfer to low-bicarbonate solution containing iron than did chlorotic plants pretreated with low-carbonate solution lacking iron. Initial localization of iron occurred in the roots. A considerable amount of the iron initially found on the roots was translocated to developing shoots over a nine-week period unless the plants were grown in high bicarbonate solutions. More iron was translocated from roots of plants in minus-iron solutions following initial absorption than when iron was supplied in the nutrient solutions. Journal Series Paper736. University of Georgia, College of Agriculture Experiment Stations, College Station, Athens, Ga. 30601.