Factors related to iron uptake by dicotyledonous and monocotyledonous plants III. Competition between root and external factors for Fe

In comparison studies (11, 12), monocotyledonous corn (Zea mays L.) and oats (Avena byzantina C. Koch) did not respond to Fe stress as effectively nor to the same degree as the dicotyledonous soybeans (Glycine max (L.) Merr.) or tomatoes (Lycopersicon esculentum Mill.). Both the Fe‐inefficient and Fe‐efficient corn and oats developed Fe chlorosis; the Fe‐efficient dicotyledonous plants were green. In the present study, the method of inducing Fe stress was changed to make it less severe. Instead of using only NO₃‐N and no Fe to induce Fe stress (11, 12), both NH₄‐N and NO₃‐N were used along with varied concentrations of Fe. Iron stress was induced with BPDS (4,7‐diphenyl‐l, 10‐phenan‐throline disulfonic acid) and phosphate; both competed with the plant for Fe. Phosphate also inhibits reduction of Fe³⁺ to Fe²⁺ (12). This method of inducing Fe stress in the plants was less severe than using only NO₃‐N and no Fe in the nutrient solutions and we were able to measure a difference in Fe‐stress response for all four plant species (Fe‐inefficient and Fe‐efficient). At the lower Fe treatments, the roots of Fe‐efficient plants usually reduced more Fe³⁺ to Fe²⁺ than did the roots of Fe‐inefficient plants. The ‘inefficient’ ys₁ corn and TAM 0–312 oat roots did not compete with BPDS or phosphate for Fe as effectively as did the ‘efficient’ WF9 corn and Coker 227 oat roots. The same type mechanism for solubilization, absorption, and transport of Fe seems to function in both monocotyledenous and dicotyledenous plants but it is more effective and more readily detected in the dicot than in the monocot plants. The reactions involved in reduction of Fe³⁺ to Fe²⁺ seemed to be confined inside or at the root surface for the inefficient genotypes; the efficient genotypes alter the ambient medium to a greater degree.     

Factors related to iron uptake by dicotyledonous and monocotyledonous plants. I. pH and reductant

Some plants respond to Fe‐deficiency stress by inducing Fe‐solubilizing reactions at or near the root surface. In their ability to solubilize Fe, dicotyledonous plants are more effective than monocotyledonous plants. In this study we determined how representative plants differ in their response when subjected to Fe‐deficiency stress in a calcareous soil and in nutrient solutions. Iron‐inefficient genotypes of tomato, soybean, oats, and corn all developed Fe chlorosis when grown in soil, whereas Fe‐efficient genotypes of these same species remained green. The same genotypes were grown in complete nutrient solutions and then transferred to nutrient solutions containing N (as NO₃ ^‐) and no Fe.

The T3238 FER tomato (Lycopersican esculentum Mill.) Fe‐efficient) was the only genotype that released significant amounts of H from the roots (the pH was lowered to 3.9) and concomitantly released reductants. Under similar conditions, Hawkeye soyhean [Glycine max (L.) Merr.] released reductants but the solution pH was not lowered. Both Fe‐inefficient and Fe‐efficient genotypes of oats (Avena sativa L.) and corn (Zea mays L.) released insufficient H or reductant from their roots to solubilize Fe; as a result, each of these genotypes developed Fe‐deficiency (chlorosis).

The marked differences observed among these genotypes illustrate the genetic variability inherent within many plant species. A given species or genotype may accordingly not be adapted to a particular soil. Conversely, a given species or genotype may be found (or developed) that is precisely suited for a particular soil. In this event, the need for soil amendments may be reduced or eliminated.     

Influences of ultra‐violet (UV)‐blue light radiation on the growth of cotton. I. Effect on iron nutrition and iron stress response

Cool white fluorescent (CWF) light reduces Fe³⁺ to Fe²⁺ while low pressure sodium (LPS) light does not. Cotton plants grown under CWF light are green, while those yrown under LPS light develop a chlorosis very similar to the chlorosis that develops when the plants are deficient in iron (Fe). It could be that CWF light (which has ultra violet) makes iron more available for plant use by maintaining more Fe²⁺ in the plant. Two of the factors commonly induced by Fe‐stress in dicotyledonous plants‐‐hydroyen ions and reductants released by the roots‐‐were measured as indicators of the Fe‐deficiency stress response mechanism in M8 cotton.

The plants were grown under LPS and CWF light in nutrient solutions containing either NO₃‐N or NH₄‐N as the source of nitrogen, and also in a fertilized alkaline soil. Leaf chlorophyll concentration varied significantly in plants grown under the two light sources as follows: CWF+Fe > LPS+Fe > CWF‐Fe ≥ LPS‐Fe. The leaf nitrate and root Fe concentrations were significantly greater and leaf Fe was generally lower in plants grown under LPS than CWF light. Hydrogen ions were extruded by Fe‐deficiency stressed roots grown under either LPS or CWF light, but “reductants”; were extruded only by the plants grown under CWF light. In tests demonstrating the ability of light to reduce Fe³⁺ to Fe²⁺ in solutions, enough ultra violet penetrated the chlorotic leaf of LPS yrown plants to reduce some Fe³⁺ in a beaker below, but no reduction was evident through a yreen CWF grown leaf.

The chlorosis that developed in these cotton plants appeared to be induced by a response to the source of liyht and not by the fertilizer added. It seems possible that ultra violet liyht could affect the reduction of Fe³⁺ to Fe²⁺ in leaves and thus control the availability of this iron to biological systems requiring iron in the plant.     

Root responses of sterile‐grown onion plants to iron deficiency 1

Onion (Allium sativum) plants grown without iron (Fe) in sterile nutrient solutions readily developed chlorosis symptoms. Iron deficiency in the sterile‐grown plants stimulated the rates of root extracellular reduction of Fe³⁺, copper (Cu²⁺), manganese (Mn⁴⁺), and other artificial electron acceptors. While rapid reduction occurred with the synthetic chelate Fe³⁺HEDTA, no short‐term reduction occurred with the fungal siderophore Fe³⁺ferrioxamine B (FeFOB). In addition to the increased rate of extracellular electron transfer at the root surfaces, the Fe‐deficient plants showed greater rates of Fe uptake and translocation than the onion plants grown with Fe. The rates of uptake and translocation of Fe were sharply higher for the Fe‐deficient plants supplied with FeHEDTA than for similar plants supplied with FeFOB. Inhibition by BPDS of the Fe uptake by the Fe‐deficient onion plants further supported the importance of Fe³⁺ chelate reduction for the uptake of Fe into the roots. Rates of Fe uptake and translocation by Fe‐deficient onion plants supplied with ⁵⁵FeFOB were identical to the rates of uptake of ferrated [14C]‐FOD; a result that gives evidence of the uptake and translocation of the intact ferrated siderophore, presumably by a mechanism not involving prior extracellular Fe³⁺ reduction. Differences in the rates of transport of other micronutrients into the roots of the Fe‐deficient onion plants were evident by the significantly higher Zn and Mn levels in the shoots of the Fe‐deficient onion.     

Micronutrient deficiencies in rainfed calcareous soils of Pakistan. I. Iron chlorosis in the peanut plant

Abstract

Peanut (Arachis hypogaea L.) is susceptible to iron (Fe) chlorosis, however, plant analysis diagnostic criteria are lacking for determining the intensity of chlorosis in this crop. As total Fe content is a misleading index of Fe nutritional status of plants, determination of physiologically active Fe fraction (Fe²⁺) is suggested for the purpose. In a nutrient indexing survey of the chlorosis‐affected peanut crop grown in the rainfed Potohar plateau of Pakistan, o‐phenanthroline extractable Fe²⁺ concentration in plants decreased with increasing severity of chlorosis and thus proved an effective technique for determining the intensity of Fe chlorosis. Green plants contained 40.1 to 67.3 mg Fe²⁺/kg, mildly chlorotic 32.1 to 40.0 mg Fe²⁺/kg, moderately chlorotic 28.0 to 32.0 mg Fe²⁺/kg, and severely chlorotic <28.0 mg Fe²⁺/kg. The minimum Fe²⁺ requirement in green plants was estimated to be 40 mg/kg on dry weight basis. In rainfed field experiments on a calcareous Typic Hapludalfs soil, foliar sprays of 1% solution of sequestrene (NaFeEDDHA) proved superior to the foliar sprays of 0.5% FeSO₄.7H₂O in correcting Fe chlorosis in two cultivars of peanut. Maximum increase in pod yield with sequestrene was 42% in cv. BARD‐92 and 27% in cv. BARD‐699 over the respective control yields. Ferrous concentration in plants increased with both the Fe sources, however, a substantial increase was recorded only with sequestrene. As peanut is a low‐input high‐risk rainfed crop, correction of Fe chlorosis by using sequestrene may not be economically feasible. Thus, development and/or screening of peanut varieties tolerant to Fe chlorosis is suggested by employing Fe²⁺ analysis technique.     

Effect of bicarbonate on iron reduction by soybean roots 1

Release of reducing compounds by soybean (Glycine max (L.) Merr.] roots has been identified as an adaptive response mechanism to iron‐deficiency conditions which result in chlorosis. These compounds facilitate the conversion of Fe⁺³ to the metabolically active Fe⁺² form, allowing for increased uptake by roots in solution culture experiments. Degree of chlorosis is closely associated with HCO₃ concentration; however, the relationship between that ion and root reduction potential apparently has not been studied. We examined the effect of HCO₃‐ on root reduction potential of ten commercially‐grown soybean cultivars known to differ in chlorosis expression in the field. Root reduction potential was measured spectrophotometrically at 594 nm on samples of nutrient solution containing reduced Fe⁺² . Plants were grown with 5 mM NaHCO₃ or in HCO₃ ^‐‐free solutions. Averaged over cultivars, 0.205 umoles Fe⁺³ were reduced in the HCO₃ ^‐‐free solutions while only 0.009 umoles Fe⁺³ were reduced in the solutions containing HCO₃ ^‐. No significant differences were observed among cultivars for root reduction potential within either HCO₃ ^‐ treatment. Results from this study suggest that HCO₃ ^‐ may inhibit iron absorption by limiting the ability of roots to release reducing compounds which make available Fe⁺2 in the soil solution. This may partially explain the role of HCO₃ ^‐ in reducing chlorosis.     

Iron inefficient and efficient oat cultivars. II. Characterization of phytosiderophore released in response to Fe deficiency stress

Abstract

Iron‐inefficient TAM 0–312 and Fe‐efficient Coker 227 oats (Strategy II plants) differ in their release of phytosiderophore in response to iron‐deficiency stress—the Fe‐efficient Coker 227 releases a phytosiderophore whereas the Fe‐inefficient TAM 0–312 does not. The phytosiderophore released by Coker 227 oats in response to Fe‐deficiency stress does not appear to transport Fe into the plant as Fe phytosiderophore. When the Fe‐inefficient TAM 0–312 and Fe‐efficient Coker 227 oats were subjected to Fe supplied as Fe²⁺(BPDS)₃, Fe³⁺HEDTA, as Fe³⁺EDDHA, Coker 227 utilized the Fe more efficiently than TAM 0–312 in every case. Both cultivars reduced Fe³⁺ as FeCl₃ to form Fe²⁺(BPOS)₃ and responded better to this form of Fe than Fe supplied as the ferric chelate. Reduction of Fe³⁺ at the root appears to be a factor that facilitates iron uptake by Coker 227 oats and the release of a phytosiderophore appears to make more Fe available at the root that can be reduced and transported to plant tops.     

Tumorous crown gall response to iron‐deficiency stress in sunflower

Tumorous crown gall tissue in sunflower (Helianthus annus L.) initiates a mechanism for making Fe available to itself as evidenced by its ability to reduce Fe³⁺ to Fe²⁺. The objective of this study was to determine if a limited Fe supply to the plant might affect the growth, nutrition and reduction of Fe³⁺ to Fe²⁺ by the tumorous crown gall. Healthy green 14‐day‐old sunflower plants (cv mammoth Russian) were either stem‐inoculated with Agrobacterium tumefaciens to induce tumorous crown gall tissue development or were left uninoculated for comparison. The plants were grown in a modified Hoagland nutrient solution with treatments containing 0.0, 0.15, 0.6 and 2.0 mg Fe L^‐1. The 0 mg Fe L^‐1 treatment induced maximum Fe chlorosis, and consequently there was a release of hydrogen ions and of a yellow pigment by the roots, but there was no measureable release of ‘reductants’ by the roots. Iron‐deficiency stress (0 mg Fe L^‐1) also resulted in reduced tumorous crown gall growth, less reduction of Fe³⁺ to Fe²⁺, and lower levels of Fe in the tumorous tissue compared to tumorous tissues adequately supplied with Fe. The tumorous crown gall tissue on the stem reduced much more Fe³⁺ to Fe²⁺ than the nontumorous stem tissue regardless of Fe level in the treatment. Tumor tissue contained more Fe, Cu and P than the nontumorous stem tissues which may indicate a modified metabolism in this tissue. An abundant supply of Fe seems to enhance the development and growth of the tumorous crown gall tissue and a deficient supply of Fe retards its growth.     

Soybean response to iron‐deficiency stress as related to iron supply in the growth medium

The Fe‐inefficient T203 and the Fe‐efficient A7 and Pioneer 1082 (P1082) soybeans (Glycine max (L.) Merr.) were grown hydroponically with no (0 mg Fe L^‐1 ; ‐Fe) and a minute level (0.025 mg Fe L^‐1 ; +Fe) of Fe to (a) compare their responses to Fe‐deficiency stress and (b) relate Fe‐efficiency in soybeans to their ability to initiate the Fe‐stress‐response mechanism at low levels of Fe. With no Fe in solution, P1082 released similar levels of H⁺ ions, but released less reductant from their roots and there was less reduction of Fe³⁺ to Fe²⁺ by their roots than by A7 roots. These responses were also one day later and occurred after a more severe chlorosis and a lower leaf Fe had developed in P1082 than in A7. With 0.025 mg L^‐1 of solution Fe, it was not necessary for the Fe‐stress response mechanism to be fully activated to make Fe available in A7 soybean, whereas a strongly enhanced Fe stress response was observed in P1082. Increased Fe uptake and regreening of leaves immediately succeeded initiation of the Fe stress response in both cultivars and at both levels of Fe. Thus, P1082 was slightly less efficient than A7 soybean, but would be classed more efficient than the previously studied soybean cultivars A2, Hawkeye, Bragg, Pride, Anoka, and T203. These results support the hypothesis that the most efficient soybeans are those which can initiate the Fe‐stress response mechanism with little or no Fe in the growth medium. The near simultaneous occurrence of the factors in the Fe‐stress response mechanism (H ion and reductant release, reduction of Fe to Fe by roots), and the immediate increase in leaf Fe and chorophyll contents following that response suggest that all these factors act in concert, not independently, to aid in the absorption and transport of Fe to plant tops.     

Iron treatment of lime‐induced chlorosis: Implications for chlorophyll,Fe2 +, Fe3 + and K+ in leaves 1

This study addressed some complementary aspects related to plant Fe nutrition. A field and a greenhouse experiment were conducted to monitor changes in chlorophyll, Fe³⁺, Fe²⁺, Ca²⁺ and K⁺ along with the progressive evolution of lime‐induced chlorosis, and following soil (Fe‐EDDHA, Fe‐EDTA, Fe‐DTPA, DTPA) and foliar (Fe‐EDDHA, FeSO₄, “Fe‐Metalosate") treatments, in a chlorosis‐susceptible ornamental plant, Hydrangea macrophylla, over a year's growing period. Though soil Fe‐EDDHA was the most effective compound in alleviating chlorosis symptoms, it became less so with time and was only partly effective as a foliar spray. Leaf analysis showed that as chlorosis intensified and chlorophyll content decreased, phenanthroline ‐ Fe (Fe²⁺) decreased with corresponding increases in total iron (Fe³⁺) and K⁺ concentrations. The reliability of these chlorosis‐indicators was confirmed as the reverse changes occurred upon chlorosis plant recovery.     

Light‐induced reduction of FE3+ as related to causes of chlorosis in cotton

In growth chambers, cotton (Gossypium hirsutum L. genetic selection ‘M8') was grown in a synthetic growth medium under four light regimes: low pressure sodium (LPS), LPS + Incandescent (Inc), cool white fluorescent (CWF) and CWF + Inc lamps at 22 C under LPS lamps. Less chlorosis developed at 26 C than at 22 C and less under LPS + Inc than under LPS lamps. All plants were green under CWF and CWF + Inc light. Green and chlorotic plant tissue contained about the same concentrations of Fe. The proposed hypothesis was that chlorotic tissue’ contained less Fe²⁺ than green tissue. Chlorotic leaves treated with FeSO₄ turned a green color. Enough CWF + Inc light passed through an intact leaf to reduce Fe³⁺ to Fe²⁺ in vitro. Also in vitro, Fe³⁺ was reduced by CWF, by Inc, but not by LPS light. The amount of Fe³⁺ reduced during an illumination period was directly proportional to the quantity of light used. In vitro, citrate and malate enhanced Fe³⁺ reduction, whereas phosphate, pyrophosphate, OH^‐, Cu²⁺, Ni²⁺, Mn²⁺, Zn²⁺, and F^‐ all inhibited Fe³⁺ reduction by light. Orthophosphate was about 8 times as effective as organic P in decreasing Fe³⁺ reduction. Citrate largely alleviated the inhibitory effects of Pi and pH (up to pH 6). The data also provide a possible explanation of a role for many of the elements known to induce or aggravate Fe chlorosis (inhibit Fe³⁺ reduction). Quantity and quality of light apparently play key roles in plant growth as related to reduction of Fe³⁺ to Fe²⁺ in plant tops.     

Tumorous crown gall response to iron‐deficiency stress in Fe‐efficient T3238fer and Fe‐inefficeent T3238fer tomatoes

Abstract

Although sunflower (Helianthus annus L.) is an Fe efficient plant, tumorous crown gall tissue development and tissue ability to reduce Fe³⁺ to Fe²⁺ were both diminished by Fe‐deficiency stress. Crown gall also develops readily on Fe‐efficient and Fe‐inefficient tomato cultivars (Lycopersicon esculentum Mill.). The objective of this study was to determine if the effect of a limited Fe supply on the growth, nutrition and reduction of Fe³⁺ to Fe²⁺ by tumorous crown gall would differ between Fe‐efficient T3238FER and Fe‐inefficient T3238fer tomato. Healthy green 25‐day‐old plants were either stem‐inoculated with Agrobacterium tumefaciens to induce tumorous crown gall tissue development or were left uninoculated for comparison. Plants were grown in modified Hoagland nutrient solutions containing 0.0, 0.15, 0.6 and 2.0 mg Fe L^?1. Yield of tumorous crown gall tissue was not diminished by low solution Fe in T3238FER, but was in T3238fer. This was attributed to inability of the T3238fer tomato to make Fe available to itself. Tumor tissue from both cultivars contained more Fe, Cu and P than normal stem tissues, which confirms a modified metabolism in these tissues previously observed in sunflower. An abundant supply of Fe enhances the development and growth of the tumorous crown gall tissue, but a deficient supply of Fe retards its growth.     

Modified procedures for commercial adaptation of root iron‐reducing capacity as a screening technique

Abstract

Genotypic evaluation is critical to development of soybean [Glycine max (L.) Merr.] cultivars with genetic resistance to Fe‐deficiency chlorosis. Root Fe³⁺ reducing activity is correlated with genotypic resistance to Fe chlorosis measured in field nurseries, and thus may be a reliable method for identifying chlorosis‐resistat genotypes. However, to develop methods useful for large‐scale screening, several modifications of the previously published procedure for measuring root Fe³⁺ reducing activity were investigated. Several hydroponic experiments were conducted to test proposed modifications. It was determined that: (a) different genotypes may be grown together in the same nutrient solution without affecting Fe³⁺ reduction, (b) genotype separation is maximized by growth in CaCO₃ buffered solution (37.5 mg L^?1), (c) a labor‐intensive elongation step can be eliminated, and (d) denotype evaluation can be accomplished without introducing Fe into the hydroponic solutions. These refinements to the procedure should allow its adaptation and use in soybean breeding programs.     

柑桔根系对缺铁的生理反应

柑桔在缺铁条件下,幼苗根系分泌H⁺和还原Fe³⁺的能力增加,与植株吸收铁素营养的能力相一致。摘除缺铁失绿的新叶和顶芽,抑制根系分泌H⁺和Fe³⁺还原物;但随着新叶的重新长出,根系分泌H⁺和Fe³⁺还原物的速率逐渐提高。说明缺铁失绿的组织能调控根系分泌H⁺和Fe³⁺还原物。在营养液中加入0.1nmol/L生长素(IAA),能解除因缺少失绿组织对根系分泌H～+产生的抑制作用。此外,在缺铁条件下,吸铁能力较强的酸橙根尖发生膨胀,有利于铁的接触吸收。     

Chelation effects on the iron reduction and uptake by low‐iron stress tolerant and non‐tolerant citrus rootstocks

The relative rates of ferric‐iron (Fe³⁺) reduction and uptake by two citrus rootstocks were measured for a series of synthetic Fe³⁺ chelates and microbial siderophores. The rates of Fe³⁺ reduction by the citrus seedlings followed the order: FeHEDTA >> FeDPTA > FeCDTA. No reduction occurred for FeDFOB (ferrioxamine B) and FeTAF (ferric triacetylfusagen). Low rates of Fe³⁺ reduction occurred for Fe2RA3 (ferric rhodotorulic acid). The levels of ⁵⁵Fe taken up the citrus seedlings showed good correlations with the reduction rates. These results indicate the importance of Fe³⁺ reduction in the Fe uptake by citrus rootstocks. The immobility of a large percent of the ⁵⁵Fe taken up by the roots is attributed to the accumulation of Fe in the root apoplasts.     

Iron deficiency studies of sugar beet using an improved sodium bicarbonate‐buffered hydroponic growth system

A sodium bicarbonate (NaHCO₃)‐buffered hydroponic growth system was developed that simulates alkaline soil growth conditions necessary to screen sugar beet genotypes for iron (Fe) efficiency character. Three genotypes (NB1, NB4, and F, hybrid, NB 1xNB4) with differing capacities for Strategy I Fe responses were phenotyped successfully using this system. Genotypes NB1 and NB1xNB4 are Fe efficient, while NB4 is Fe inefficient. It was demonstrated that 5 mM NaHCO₃ provided buffering within an optimal range (pH 7.3 ‐ pH 6.3) for the duration of ‐Fe treatments, promoted enhanced H⁺ extrusion, and increased the in vivo capacity for Fe³⁺‐chelate reduction (Fe³⁺‐chelate reductase [FCR] activity), especially in the roots of the Fe efficient genotypes. The same concentrations of NaHCO₃ did not interfere with Fe supply to +Fe control plants of any genotype. The in vivo capacity for Fe³⁺‐chelate reduction increased over fivefold in both Fe efficient genotypes (NB1 and NB 1xNB4), but just under twofold in the Fe inefficient genotype (NB4). Localization and duration of enhanced Fe³⁺‐chelate reduction capacity were dependent upon the Fe efficiency character of each genotype.     

An iron chelating compound released by barley roots in response to Fe‐deficiency stress

The excretion of phytosiderophores by barley (Hordeum vulgare L.) has recently been documented and a major difference in the Fe‐stress response of gramineous species and dicotyledonous species proposed. However, currently used methods of quantifying and measuring phytosiderophore are tedious or require specialized equipment and a cultivar easily accessible to U.S. scientists is needed. The objectives of this study were (a) to determine if “Steptoe”; and “Europa”; (used as a control cultivar) barleys would release Fe³⁺ solubilizing compounds in response to Fe‐deficiency stress and (b) to develop a technique to determine the efficiency of solubilization of Fe(OH)₃ by the released chelating substances. Two cultivars of barley were place under Fe‐stressed (‐Fe) and nonstressed (+Fe) conditions in modified Hoagland solutions (14 L). The solutions were periodically monitored for H⁺ and reductant release from the roots and plants were rated daily for chlorosis development. Periodic (6 or 7 harvests) evaluation of the release of Fe³⁺ solubilizing substances was performed as herein described. Neither H⁺ nor reductant extrusion occurred with either cultivar during Fe stress. However, Fe³⁺ solubilizing substances were released by both cultivars at relatively high levels under Fe‐stress conditions compared to the nonstressed plants. A convenient technique was developed to measure the release of Fe solubilizing substances released by barley roots.     

Correction of iron chlorosis in peanut (arachis hypogea shulamit) by ammonium sulfate and nitrification inhibitor

Peanut (Arachis hypogea cv. Shulamit) grown on very high calcium carbonate (CaCO₃) content soils is showing iron (Fe) chlorosis symptoms. Supplying the plant with ammonium sulphate ((NH₄)₂SO₄) in the presence of nitrapyrin (N‐Serv) for preventing nitrification reduced Fe chlorosis. Nitrate (NO^‐ ₃) developed in the soil with time, even with nitrapyrin present. When ammonium (NH⁺ ₄) was even less than 20% of the total mineral N in the soil, no Fe‐stress could be observed, suggesting that the NH⁺ ₄ uptake by the plant and the consequence of hydrogen (H⁺) efflux occurs from the root to the rhizosphere, resulting in a decrease of redox potential near the root, and solubilizing enough Fe near the root to overcome the chlorosis.     

Physiological responses of grapevine callus cultures to iron deficiency

Grapevine is considered a ‘Strategy I’ plant because it performs some peculiar biochemical and physiological responses when grown under iron (Fe) deficiency stress conditions. Callus cultures were started from leaf and internode cuts of micropropagated plantlets of two grapevine genotypes well known for their Fe‐chlorosis characteristic: Vitis riparia a very susceptible genotype and Vitis berlandieri a resistant one. Modification of NADH: ferric (Fe³⁺) reductase activity was spectrophotometrically evaluated by following the formation of the complex ferrous (Fe²⁺)‐(BPDS)₃, while the malic and citric acid production were determined in callus cultures grown both in the presence (+Fe) and absence (‐Fe) of Fe. Moreover, a microsomal fraction was isolated from the calli to evaluate the H⁺‐ATPase and the Fe³⁺‐EDTA reductase activities. As expected, calli of the Fe‐efficient genotype (V. berlandieri) was able to enhance Fe³⁺‐EDTA reductase activity when growing under Fe deficiency while the Fe‐chlorosis susceptible V. riparia could not or did it with lower efficiency. Therefore, the H⁺‐ATPase assay showed a higher enzymatic activity in the microsomal fraction isolated from Vitis berlandieri grown without Fe with respect to its control (+Fe). Organic acid determination gave quite contradictory results, specially regarding malic acid which, under our study conditions, seemed not to be linked with the strategies of response to Fe deficiency.     

Factors in iron‐stress response mechanism enhanced by Zn‐deficiency stress in Sanilac,but not Saginaw navy bean

Zinc‐inefficient Sanilac and Zn‐efficient Saginaw navy bean (Phaseolus vulgaris L.) differ in their susceptibility to Zn‐deficiency stress. Sanilac accumulates Fe under Zn‐deficiency stress and Saginaw does not. These two navy bean cultivars were grown at 0, 0.006 and 0.12 mg/L Zn in modified Hoagland nutrient solution. Various Fe‐stress response mechanisms were quantified periodically over a 12‐day experimental period to determine if known factors in the Fe‐stress response mechanism were enhanced by Zn‐deficiency stress. Visual Zn‐deficiency symptoms were more severe in Sanilac than Saginaw navy bean under equivalent Zn treatments. Sanilac contained lower leaf Zn than Saginaw when Zn was present in solution (0.006 and 0.12 mg/L Zn), but the two cultivars were similar in leaf Zn in the absence of Zn (0 mg/L Zn). Sanilac accumulated more leaf Fe than Saginaw when under Zn stress (0 and 0.006 mg/L Zn). The higher levels of leaf Fe in Sanilac than Saginaw were closely associated with enhanced release of reductants and increased reduction of Fe³⁺ to Fe²⁺ by roots of Sanilac. Saginaw navy bean roots reduced Fe³⁺ to Fe²⁺ similarly to Sanilac with adequate Zn present in solution (0.12 mg/L), but experienced minuscule levels of Fe³⁺ reduction under Zn deficiency. Zinc deficiency stimulated the initiation of the Fe‐stress response mechanism in Sanilac, but not Saginaw, which may have enhanced the development of Zn‐deficiency symptoms in Sanilac due to the increased uptake of Fe by this cultivar. The common Fe‐deficiency stress response associated primarily with grasses (release of phytosiderophore) was not found in either navy bean cultivar.