IRON IS REQUIRED FOR THE INDUCTION OF ROOT FERRIC CHELATE REDUCTASE ACTIVITY IN IRON-DEFICIENT TOMATO

Two mutants of tomato and their corresponding wild-type genotypes, Tfer/TFER and chloronerva/Bonner Beste, were grown in nutrient solution under conditions leading to iron (Fe) deficiency. Iron deficiency caused decreases in growth, leaf chlorosis, and changes in the morphology of roots. Ferric chelate reductase activities of whole roots were generally lower in Fe-deficient plants than in control, Fe-sufficient plants. Plants grown for 7 days without Fe, however, had transient increases in whole root ferric chelate reductase activity after the addition of small amounts of Fe (2 μM) to the nutrient solution. Also, adding sequential 0.5 μM Fe pulses to the nutrient solution led to high whole root ferric chelate reductase activities. Similar results were obtained with a protocol using excised root tips instead of whole root systems to measure ferric chelate reductase activities. The protocol using root tips generally gave higher ferric chelate reductase rates than the method using whole roots, due to the localized expression of the enzyme in the distal root zones.     

Iron deficiency responses in roots of kiwi

Many dicotyledonous species respond to iron (Fe) deficiency by morphological and physiological changes at root level, which are usually defined as Strategy I. Particularly, these latter modifications include a higher acidification of the external medium and the induction of a high root Fe reductase activity. The aim of this work was to investigate the response of kiwi (Actinidia deliciosa cv. Hayward) plants, which often exhibit Fe chlorosis in the field, to Fe deficiency. Actinidia kept for two weeks in nutrient solution without Fe showed visual deficiency symptoms (leaf chlorosis). Moreover, upon prolonged micronutrient shortage shoot, and to a lesser extent, root dry weight accumulation was greatly impaired. Roots of Fe‐deficient Actinidia showed an increased capacity of net proton extrusion and higher ferric ethylenediaminetetraacetate [Fe(III)EDTA] reductase activity as compared to plants grown in the presence of 10 μM Fe(III)EDTA. Localization of the increased acidification and reductase capacity by means of agar‐technique revealed that these activities are both present in the sub‐apical region of the roots. Re‐supply of Fe after two weeks partially reversed the tendency of the roots to acidify the nutrient solution and to reduce Fe(III)EDTA.     

RESPONSES OF “NEWHALL” ORANGE TREES TO IRON DEFICIENCY IN HYDROPONICS: EFFECTS ON LEAF CHLOROPHYLL,PHOTOSYNTHETIC EFFICIENCY,AND ROOT FERRIC CHELATE REDUCTASE ACTIVITY

Orange (Citrus sinensis L. Osb. cv. ‘Newhall’) plants grafted on Citrange troyer rootstock were grown in nutrient solution with 0, 5, 10, or 20 μM iron (Fe), with and without calcium carbonate. Calcium carbonate was added in order to mimic the natural conditions in calcareous soils. Leaf chlorophyll concentration was estimated every 3–4 days using the portable instrument SPAD-502 meter. Chlorophyll fluorescence parameters, photosynthetic capacity estimated from oxygen evolution, leaf Fe concentrations, and root tip ferric chelate reductase activity were measured at the end of the experiment. Plants from the 0 and 5 μM Fe treatments showed leaf chlorosis and had decreased leaf chlorophyll concentrations. Leaves of plants grown in the absence of Fe in the solution had smaller rates of oxygen evolution both in the presence and absence of calcium carbonate, compared with plants grown in the presence of 10 μM Fe. In the absence of calcium carbonate the photosystem II efficiency, estimated from fluorescence parameters, was similar in all treatments. A slight decrease in photosystem II efficiency was observed in plants grown without Fe and in the presence of calcium carbonate. A 2.5-fold increase in root tip ferric chelate reductase activity over the control values was found only when plants were grown with low levels of Fe and in the presence of calcium carbonate.     

RELATIVE SUSCEPTIBILITY OF SOME PRUNUS ROOTSTOCKS IN HYDROPONICS TO IRON DEFICIENCY

The susceptibility of eleven Prunus rootstocks to iron (Fe) deficiency was studied in hydroponics by growing them with 20 μM Fe, 0 μM Fe or 3 μM Fe+10 mM sodium bicarbonate. Based on the intensity of leaf chlorosis, the peach-almonds PR 204/84, Stylianidis K and KID2, produced at the Pomology Institute of Naoussa (Greece), showed the same or even greater tolerance than GF 677, the Greek peach-almond Retsou x Nemaguard, the plum-almond Myrandier 617 and the peaches GF 305, IDS 37, Greek wild peach seedling the greatest susceptibility whereas the plums St. Julien GF655/2 and Myrobalan 29C intermediate. Rootstocks without Fe presented significantly lower nitrogen and Fe whereas with bicarbonate significantly lower nitrogen, phosphorus, Fe and zinc. Root ferric chelate reductase activity was significantly increased in ?Fe rootstocks but negatively correlated with their tolerance; physiological and morphological changes were observed along a zone of a few centimeters length, 1–2 mm behind the root tip.     

Root and Leaf Ferric Chelate Reductase Activity in Pond Apple and Soursop

Abstract

Root and leaf ferric chelate reductase (FCR) activity in Annona glabra L. (pond apple), native to subtropical wetland habitats and Annona muricata L. (soursop), native to nonwetland tropical habitats, was determined under iron (Fe)-sufficient and Fe-deficient conditions. One-year-old seedlings of each species were grown with 2, 22.5, or 45 µM Fe in a nutrient solution. The degree of tolerance of Fe deficiency was evaluated by determining root and leaf FCR activity, leaf chlorophyll index, Fe concentration in recently mature leaves, and plant growth. Root FCR activity was generally lower in soursop than in pond apple. Eighty days after plants were put in nutrient solutions, leaf FCR activity of each species was lower in plants grown with low Fe concentrations (2 µM) than in plants grown with high (22.5 or 45 µM) Fe concentrations in the nutrient solution. Leaves of pond apple grown without Fe became chlorotic within 6 weeks. The Fe level in the nutrient solution had no effect on fresh and dry weights of soursop. Lack of Fe decreased the leaf chlorophyll index and Fe concentration in recently matured leaves less in soursop than in pond apple. The rapid development of leaf chlorosis in low Fe conditions and low root and leaf FCR activities of pond apple are probably related to its native origin in wetland areas, where there is sufficient soluble Fe for adequate plant growth and development. The higher leaf FCR activity and slower growth rate of soursop compared to pond apple may explain why soursop did not exhibit leaf chlorosis even under low Fe conditions.     

Strawberry recovers from iron chlorosis after foliar application of a grass‐clipping extract

Bare‐root transplants of strawberry (Fragaria × ananassa Duch. cv. Selva) were transferred to nutrient solutions with or without iron. After 35 d of growth, plants in the solution without iron became chlorotic and had morphological changes in roots typical of iron‐deficiency chlorosis (IDC). Acidification of the nutrient solution was also observed. We tested a grass‐clipping extract to correct IDC in strawberry plants by foliar application to some chlorotic plants. We also assessed the effects of this product on plant growth, Fe allocation, as well as morphological and physiological parameters related with IDC. After the second spray, leaf chlorophyll increased in the youngest expanded leaves. The total content of iron in plants increased from 1.93 mg to 2.37 mg per plant after three sprays, accounting for 80% of the total iron supplied by the extract. Newly formed roots from sprayed plants had a normal morphology (no subapical swollen zone) but a higher ferric chelate–reductase (FC‐R; EC 1.16.1.17) activity per root apex compared with roots from plants grown with iron or untreated chlorotic plants. Acidification of the nutrient solution continued in sprayed recovered plants. The results suggest an uncoupling of the regulation of morphological and physiological mechanisms related to IDC: FC‐R activity seems to be controlled by roots on their own or together with shoots, while morphological changes in roots are apparently regulated only by the level of iron in shoots.     

Influence of Ca concentration on micronutrient imbalances in in vitro propagated prunus rootstock

In vitro propagated plums of St. Julien GF 655–2 [Prunus insititia (L.)] (655–2), Damas GF 1869 [Prunus domestica (L.)] (D1869), and Clark Hill Redleaf [Prunus salicina (Until.) X Prunus cerasifera (Ehrh.)] (CHR), were grown in the greenhouse in nutrient solutions of 2, 6, 22, 66, 202, and 404 μM Ca for 96 days. 655–2 plants became severely chlorotic in Ca treatments of 66, 202, and 404 μM concentration after 86 days of growth. D1869 plants in 202 and 404 μM Ca exhibited slight interveinal chlorosis of new growth, while CHR exhibited no chlorosis at any Ca concentration. The best tissue nutrient indicator of chlorosis among rootstocks was foliar P/Fe and P/Zn ratios. 655–2 plants absorbed more P at higher Ca concentrations than did the other rootstock, resulting in the highest stem and leaf P/Fe, and P/Zn ratios. CHR plum may provide an easily propagated, chlorosis‐resistant rootstock for use on calcareous soils.     

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.     

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.     

Response to iron‐deficiency stress of pear and quince genotypes 1

Roots of iron (Fe)‐efficient dicots react to Fe‐deficiency stress by strongly enhancing the ferric (Fe³⁺)‐reductase system and by lowering the rhizo‐sphere pH. In this study, we tested whether such adaptation mechanisms characterize pear and quince genotypes known to have differential tolerance to calcareous and alkaline soils. Two trials were performed using micropagated plants of three quince rootstocks (BA29, CTS212, and MC), three Pyrus communis rootstocks (OHxF51 and two selections obtained at the Bologna University: A28 and B21) and of two pear cultivars (Abbé Fétel and Bartlett, own‐rooted). In the first trial, plants were grown in a nutrient solution with [Fe(+)] and without [Fe(‐)] Fe for 50 days. Their root Fe‐reducing capacity was determined colorimetrically using ferrozine and FeEDTA, and Fe uptake of Fe(+) plants was estimated. In the second trial, the rhizosphere pH of plants grown in an alkaline soil was measured by a micro‐electrode. With the only exception of pears OHxF51 and A28, whose Fe‐reduction rates were similar in Fe(+) and Fe(‐) plants, the Fe‐deficiency stress resulted in a significant decrease in Fe reduction. Among the Fe(‐) plants, the two pear cultivars, OHxF51 and A28, had a higher Fe‐reducing capacity than the quince rootstocks and the cv. Abb6 F. When plants were pre‐treated with Fe, reduction rate was highest in the P. communis rootstocks, intermediate in the own‐rooted cultivars, and lowest in the quinces. Root Fe‐reducing capacity of Fe(+) plants proved to be linearly and positively correlated with Fe uptake and root proton release. Rhizosphere pH was highest in quince MC, intermediate in the other two quinces and in the cv. Abbe F., and lowest in the pear rootstocks and in the cv. Bartlett. Our results indicate that roots of pear and quinces do not increase their ability to reduce the Fe under Fe‐deficiency stress. The genotypical differential tolerance to Fe chlorosis likely reflects differences in the standard reductase system and in the capacity of lowering the pH at the soil/root interface. The determination of the root Fe‐reducing capacity is a promising screening technique for selecting pear root‐stocks efficient in taking up Fe.     

Activities of Iron‐Containing Enzymes in Leaves of Two Tomato Genotypes Differing in Their Resistance to Fe Chlorosis

Abstract

Two tomato (Lycopersicon esculentum Mill., cvs. Pakmor and Target) genotypes differing in resistance to iron (Fe) deficiency were grown in nutrient solution under controlled environmental conditions over 50 days to study the relationships between severity of leaf chlorosis, total concentration of Fe, and activities of Fe‐containing enzymes in leaves. The activities of Fe‐containing enzymes ascorbate peroxidase, catalase, and guaiacol peroxidase, and additionaly the activity of glutathione reductase, an enzyme that does not contain Fe, were measured. Plants were supplied with 2 × 10^?7 M (Fe deficient) and 10^?4 M (Fe sufficient) FeEDTA, respectively. Leaf chlorosis appeared more rapidly and severely in Target (Fe deficiency senstive genotype) than Pakmor (Fe deficiency resistant genotype). On day 50, Pakmor had 2‐fold more chlorophyll than Target under Fe deficiency, while at adequate supply of Fe the two genotypes were very similar in chlorophyll concentration. Despite distinct differences in development of leaf chlorosis and chlorophyll concentrations, Pakmor and Target were very similar in concentrations of total Fe under Fe deficiency. In contrast to Fe concentration, activities of Fe‐containing enzymes were closely related to the severity of leaf chlorosis. The Fe‐containing enzymes studied, especially catalase, showed a close relationship with the concentration of chlorophyll and thus differential sensitivity of tomato genotypes to Fe deficiency. Glutathione reductase did not show relationship between Fe deficiency chlorosis and enzyme activity. The results confirm that measurement of Fe‐containing enzymes in leaves is more reliable than the total concentration of Fe for characterization of Fe nutritional status of plants and for assessing genotypical differences in resistance to Fe deficiency. It appears that Fe deficiency‐resistant genotype contains more physiologically available Fe in tissues than the genotype with high sensitivity to Fe deficiency.     

Physiological responses of tomato roots grown in organ culture to iron-deficiency stress

Roots of the Fe-efficient tomato (Lycopersicon esculentum Mill., cultivar Floradel) were cultured in an inorganic medium supplemented with glycine, thiamine, pyridoxine, and nicotinic acid, with sucrose as an energy and carbon source. Iron was supplied as ferric hydroxyethylethylenediaminetriacetic acid (FeHEDTA) and the initial PH was 5.5. Root growth was limited when less than 40 μm FeHEDTA was supplied. Roots grown at lower Fe concentrations decreased the pH of the FCR assay medium to a greater extent than did roots grown at higher Fe concentrations. Cultured roots grown with 10 μm FeHEDTA had increased levels of ferric chelate reductase (FCR) activity compared to roots grown with either lower or higher concentrations of FeHEDTA. Low FCR activity of roots grown at 2.5 or 5 μm FeHEDTA was attributed either to impaired metabolism due to Fe-deficiency or the lack of sufficient Fe for enhanced FCR formation. Roots of hydroponically grown tomato plants exhibited typical increases in FCR activity with Fe-deficiency. Based on these preliminary results, cultured roots were found to exhibit similar Physiological responses to Fe-deficiency stress as intact root systems. Cultured roots should provide a useful system for the investigation of the role of the root in plant Fe-deficiency stress responses as previously suggested by Bienfait et al.(Plant Physiol., 83, 244–247, 1987).     

Response of ungrafted rootstocks and rootstocks grafted with wine grape varieties (Vitis sp.) to ‘lime-induced chlorosis’

The ungrafted rootstocks 41B, 1103P, 110R and 140Ru, the grafted combinations of 41B, 1103P and 110R with Xinomavro (one of the most important red wine grape varieties in Greece), as well as those of 1103P, 110R and 140Ru with Chardonnay, were evaluated for 'lime-induced chlorosis' tolerance by growing them with a) basic nutrient solution (BNS), b) BNS + 10 mM bicarbonate, c) BNS without iron (Fe) and d) BNS without zinc (Zn), in hydroponics. The ungrafted 140Ru followed by 41B under high bicarbonate presented the lowest degree of chlorosis; however only 41B presented non-differentiated biomass production and root/shoot ratio. Chlorotic symptoms in combination with plant growth parameters should be used as a tool for grapevine rootstock lime-tolerance screening whereas leaf Fe concentration and root ferric chelate reductase (FCR) activity should not. Lime-stress conditions affected plant mineral nutrition by depressing leaf nitrogen (N), phosphorus (P), calcium (Ca), magnesium (Mg) and increasing potassium (K), and zinc (Zn).     

Effects of Several Metals on Both Fe(III)‐ and Cu(II)‐Reduction by Roots of Fe‐Deficient Cucumber Plants

Abstract

The ferric‐chelate reductase induced by Fe deficiency is also able to reduce other ions such as Cu²⁺. This Cu(II)‐reduction has been less studied and it has been suggested that Cu²⁺ ion rather than Cu²⁺‐chelate serves as the substrate. Ferric‐chelate reductase activity is inhibited by some metals, but the mechanisms implicated are not known. In the present work we use Fe‐deficient cucumber seedlings to study the interactions of Cu²⁺, Ni²⁺, Mn⁴⁺, and Fe³⁺ on both Fe(III)‐reduction and Cu(II)‐reduction activities. The response of Cu(II)‐reduction activity to Cu concentration, in the presence or absence of citrate, was also studied. Results showed that inhibition of the ferric‐chelate reductase activity by Cu²⁺ or Ni²⁺ could be partially reversed by increasing the concentration of Fe‐EDTA. The Cu(II)‐reduction activity was even stimulated by Fe‐EDTA or Ni²⁺; it was inhibited by a high concentration of Cu²⁺ itself; and it was not affected by the absence of citrate. Mn⁴⁺ caused a moderate inhibition of both Fe(III)‐reduction and Cu(II)‐reduction activities. Results agree with the hypothesis that free Cu²⁺ ion is the substrate for Cu(II)‐reduction and suggest that the mechanisms involved in Fe(III)‐reduction and Cu(II)‐reduction could have some differences and be affected by metals in different ways. The mode of action of metals on the reductase activity are discussed, but they are still not well known.     

Nutritional (Fe-Mn) interactions in ‘Big Top’ peach plants as influenced by the rootstock and by the soil CaCO3 concentration

Manganese (Mn) toxicity in plants is often not a clearly identifiable disorder and it can interfere with the absorption, translocation, and utilization of other elements such as Ca, Mg, Fe, and P. Soil conditions, management factors, and the use of different genotypes of rootstock can determine the degree of Mn toxicity and of interaction with other elements in the orchard. Five plants of the cultivar ‘Big Top’® grafted onto itself, onto plum rootstock ‘Mr.S.2/5’ and onto hybrid peach x almond rootstock ‘GF677’ were grown in 25-L containers under three treatments, 0, 20, 30% concentration of total lime, obtained by mixing powdered CaCO₃ to a sandy soil. Plants were fertilized with manure and a solid fertilizer early in April and irrigated in summer periodically with water rich in manganese. After just 28 d, active lime caused a decrease of chlorophyll SPAD index especially in plants grafted on itself, while those grafted on the tolerant ‘GF677’ rootstock behaved better than those grafted on ‘Mr.S.2/5.’ From June to September, irrigation caused increases in soil Mn concentration and Mn concentration in control plants. This caused first a serious defoliation in Big Top / Big Top plants and then a re-greening of cultivar grafted onto ‘Mr.S.2/5’ and ‘GF677,’ probably due to the interaction between iron and manganese at high pH. In particular the 20% CaCO₃ addition to the soil preserved the plants of cultivar grafted onto ‘Mr.S.2/5’ from Mn toxicity, as shown by their high chlorophyll content and growth and lower Mn leaf concentrations. Plants grafted onto ‘GF677’ rootstock showed the best behaviour under 30% CaCO₃ treatment associated to higher Fe(III)-reducing capacity and photosynthetic activity. Rootstocks and soil conditions (lime and waterlogging) influenced mineral status and growth of the peach cultivar ‘Big Top,’ particularly by interacting together and modifying Fe-Mn availability.     

Differences in Response to Iron Deficiency Among Some Lines of Common Bean

Abstract

Five dry bean cultivars (Coco blanc, Striker, ARA14, SVM29‐21, and BAT477) were evaluated for their resistance to iron deficiency on the basis of chlorosis symptoms, plant growth, capacity to acidify the external medium and the root‐associated Fe³⁺‐reduction activity. Plants were grown in nutrient solution supplied or not with iron, 45 µM Fe(III)EDTA. For all cultivars, plants subjected to iron starvation exhibited Fe‐chlorosis. These symptoms were more severe and more precocious in BAT477 and Coco blanc than in the others cultivars. An important acidification of the culture medium was observed between the 4th and the 8th days of iron starvation in Striker, SVM29‐21 and, particularly, ARA14 plants. However, all Fe‐sufficient plants increased the nutrient solution pH. This capacity of acidification appeared more clearly when protons extrusion was measured in 10 mM KCl + 1 mM CaCl₂. The above genotypic differences were maintained: ARA14 showed the higher acidification followed by Coco blanc and BAT477. Iron deficiency led also to an increase of the root‐associated Fe(III)‐reductase activity in all lines. However, genotypic differences were observed: Striker shows the highest capacity of iron reduction under Fe deficiency condition.     

Uptake of Iron (59Fe) Complexed to Water‐Extractable Humic Substances by Sunflower Leaves

Abstract

A research was carried out to evaluate the leaves' ability to utilize Fe supplied as a complex with water‐extractable humic substances (WEHS) and the long‐distance transport of ⁵⁹Fe applied to sections of fully expanded leaves of intact sunflower (Helianthus annuus L.) plants. Plants were grown in a nutrient solution containing 10 µM Fe(III)‐EDDHA (Fe‐sufficient plants), with the addition of 10 mM NaHCO₃ to induce iron chlorosis (Fe‐deficient plants). Fe(III)‐WEHS could be reduced by sunflower leaf discs at levels comparable to those observed using Fe(III)‐EDTA, regardless of the Fe status. On the other hand, ⁵⁹Fe uptake rate by leaf discs of green and chlorotic plants was significantly lower in Fe‐WEHS‐treated plants, possibly suggesting the effect of light on photochemical reduction of Fe‐EDTA. In the experiments with intact plants, ⁵⁹Fe‐labeled Fe‐WEHS or Fe‐EDTA were applied onto a section of fully expanded leaves. Irrespective of Fe nutritional status, ⁵⁹Fe uptake was significantly higher when the treatment was carried out with Fe‐EDTA. A significant difference was found in the amount of ⁵⁹Fe translocated from treated leaf area between green and chlorotic plants. However, irrespective of the Fe nutritional status, no significant difference was observed in the absolute amount of ⁵⁹Fe translocated to other plant parts when the micronutrient was supplied either as Fe‐EDTA or Fe‐WEHS. Results show that the utilization of Fe complexed to WEHS by sunflower leaves involves an Fe(III) reduction step in the apoplast prior to its uptake by the symplast of leaf cells and that Fe taken up from the Fe‐WEHS complexes can be translocated from fully expanded leaves towards the roots and other parts of the shoot.     

Screening for resistance to iron deficiency chlorosis in dry bean using iron reduction capacity

Identifying cultivars resistant to iron (Fe) deficiency chlorosis so prevalent in calcareous soils is a more economical solution than fertilizer application in field crops. The current method of screening for resistance using chlorosis ratings in field trials is time consuming and highly variable. Root Fe reduction successfully separated cultivars or rootstocks, varying widely in resistance, of soybean (Glycine max L.), peach (Prunus persica L.), and grape (Vitis spp.), but was unsuccessful in sub‐clover (Trifolium subterraneum L.). Dry bean (Phaseolus vulgaris L.) exhibits Fe deficiency chlorosis in calcareous soils and initiates Fe reduction by the roots in response to such stress. The resistance of 24 dry bean cultivars to Fe deficiency chlorosis was assessed by measuring and summing daily Fe reduction by the roots. The cultivars were grown both hydroponically in an environmental chamber in low Fe solutions (0.05 mg‐L^‐1) and at three field sites in both 1995 and 1996. A significant relationship (P<0.01) between field chlorosis scores made 36 days after planting and root Fe reduction summations was observed for all sites in 1995 and 1996 (r = ‐0.42 to ‐0.71). The variability of chlorosis scores among sites, especially in 1996, points out the difficulty of using field chlorosis scores for screening. These results indicate that measurements of root Fe reduction can be used to predict resistance to Fe deficiency chlorosis in dry bean. Successful implementation of this technique should reduce if not eliminate field trials for screening resistance to Fe deficiency chlorosis.     

Excretion of dibutyl phthalate by sorghum roots under Fe‐stress: Evidence of its action on chlorosis recovery

Sorghum (Sorghum bicolor L. Moench) cv. CSH‐7, an Fe‐efficient hybrid was grown and subjected to Fe‐deficiency stress. The nutrient medium was extracted for isolation of reductant chemicals. By means of thin layer chromatography, I.R. spectrum and HPLC analysis, dibutyl phthalate was identified as the principal component. This chemical was not found in the nutrient medium extracted before the onset of chlorosis or in that after the plants recovered from chlorosis. Furthermore, synthetic dibutyl phthalate and that obtained from the exudate when supplied to the nutrient medium caused greening of chlorotic sorghum in 24 hours. Evidence that the root medium of the Fe‐efficient sorghum can induce recovery of an Fe‐inefficient sorghum grown together, has also been obtained. It is concluded that dibutyl phthalate released by the Fe‐efficient sorghum subjected to stress, is responsible for making Fe available for utilisation. The mechanism of action of dibutyl phthalate on chlorosis recovery is still an open question.     

Response of five citrus rootstocks to iron deficiency

Citrus established in calcareous soils can be affected by iron (Fe)‐deficiency chlorosis which limits yield and the farmers' income. The degree of deficiency depends on the rootstock, but the resistance to Fe chlorosis still requires further investigation. To study physiological parameters of citrus rootstocks that could be used to evaluate resistance to Fe deficiency, plants of Troyer citrange (Citrus sinensis L. Osb. × Poncitrus trifoliata L. Raf.), Carrizo citrange, Volkamer lemon (Citrus volkameriana Ten. & Pasq.), alemow (Citrus macrophylla Wester), and sour orange (Citrus aurantium L.) were grown in nutrient solutions with 0, 5, 10, 15, or 20 μM Fe. For each rootstock, plant height, root and shoot dry weights, and concentration of Fe in the shoots and roots were measured at the end of the experiment. Chlorophyll (CHL) concentration was estimated throughout the experimental period using a portable CHL meter (SPAD‐502) calibrated for each rootstock. At the end of the experiment, CHL fluorescence parameters were measured in each rootstock with a portable fluorimeter. Maximal and variable fluorescence values indicated that the photochemistry of Troyer was more affected by a low concentration of Fe in the nutrient solution than that of other rootstocks. To compare rootstocks, the absolute CHL concentration was converted into relative yield by employing a scaling divisor based on the maximum value of total CHL in plants without Fe‐deficiency symptoms. Exponential models were developed to determine the minimum Fe concentration in nutrient solution required to maintain leaf CHL at 50% of the maximum CHL concentration (IC50). Models were also developed to assess the period of time the rootstocks were able to grow under Fe‐stress conditions before they reached IC50. Volkamer lemon and sour orange needed the lowest Fe concentration (between 4 and 5 μM Fe) to maintain IC50, and Troyer citrange had the highest Fe requirement (14 μM Fe). Citrus macrophylla and Carrizo citrange required 7 and 9 μM of Fe, respectively. Similarly, Volkamer lemon and sour orange rootstocks withstood more days under total Fe depletion or with a low concentration of Fe (5 μM Fe in nutrient solution) until they reached IC50, compared to the other rootstocks. The approach used led to a classification of the rootstocks into three categories, regarding their internal tolerance to Fe chlorosis: resistance (sour orange and Volkamer lemon), intermediate resistance (C. macrophylla and Carrizo citrange), and reduced resistance (Troyer citrange).