Iron deficiency in peach trees: effects on leaf chlorophyll and nutrient concentrations in flowers and leaves

The effects of iron deficiency on the leaf chlorophyll concentrations and on the macro- (N, P, K, Ca and Mg) and micro-nutrient (Fe, Mn, Zn and Cu) composition of flowers (at full bloom) and leaves (60 and 120 days after full bloom) of field-grown peach (Prunus persica L. Batsch) trees were investigated. Flowers and leaves were taken and analysed from fifty individual trees. Our data indicate that large decreases in leaf chlorophyll concentration were found at the beginning of the season in control trees, possibly associated to a dilution effect by leaf growth, that were later followed by leaf chlorophyll concentration increases. Leaf Fe chlorosis apparently results from two different processes, the dilution of leaf Chl caused by growth and the subsequent inability to produce and/or stabilize new Chl molecules in the thylakoid membrane. Iron chlorosis did not change the seasonal change patterns of any of the nutrients studied. In Fe-deficient trees the K concentration and the K/Ca ratio were high not only in leaves but also in flowers, indicating that this is a characteristic of Fe-deficient plant tissue in the whole fruit tree growing season. Flower Fe concentrations were well correlated with the degree of chlorosis developed later in the season by the trees, suggesting that flower analysis could be used for the prognosis of Fe deficiency in peach.     

Metabolic response in roots of Prunus rootstocks submitted to iron chlorosis

Iron deficiency induces several responses to iron shortage in plants. Metabolic changes occur to sustain the increased iron uptake capacity of Fe-deficient plants. We evaluated the metabolic changes of three Prunus rootstocks submitted to iron chlorosis and their different responses for tolerance using measurements of metabolites and enzymatic activities. The more tolerant rootstocks Adesoto (Prunus insititia) and GF 677 (Prunus amygdalus × Prunus persica), and the more sensitive Barrier (P. persica × Prunus davidiana) were grown hydroponically in iron-sufficient and -deficient conditions over two weeks. Sugar, organic and amino acid concentrations of root tips were determined after two weeks of iron shortage by proton nuclear magnetic resonance spectroscopy of extracts. Complementary analyses of organic acids were performed by liquid chromatography coupled to mass spectrometry. The major soluble sugars found were glucose and sucrose. The major organic acids were malic and citric acids, and the major amino acid was asparagine. Iron deficiency increased root sucrose, total organic and amino acid concentrations and phosphoenolpyruvate carboxylase activity. After two weeks of iron deficiency, the malic, citric and succinic acid concentrations increased in the three rootstocks, although no significant differences were found among genotypes with different tolerance to iron chlorosis. The tolerant rootstock Adesoto showed higher total organic and amino acid concentrations. In contrast, the susceptible rootstock Barrier showed lower total amino acid concentration and phosphoenolpyruvate carboxylase activity values. These results suggest that the induction of this enzyme activity under iron deficiency, as previously shown in herbaceous plants, indicates the tolerance level of rootstocks to iron chlorosis. The analysis of other metabolic parameters, such as organic and amino acid concentrations, provides complementary information for selection of genotypes tolerant to iron chlorosis.     

Characterization of the tolerance to iron chlorosis in different peach rootstocks grown in nutrient solution

Iron chlorosis is an important problem in peach trees, but differences exist between peach rootstocks in their tolerance to Fe chlorosis in calcareous soils. The purpose of this investigation was to characterize the tolerance of different rootstocks to Fe chlorosis induced by bicarbonate in nutrient solution. The rootstocks studied included peach (Nemaguard), plums (Brompton, San Julian A and Puebla de Soto 101) and almond × peach hybrids (Adafuel and GF677). Young plants obtained from rooted cuttings or from in vitro culture techniques were grown individually, under controlled conditions, in flasks with 700 mL of aerated nutrient solution low in iron and with or without 10 mM bicarbonate or 10 mM phosphate. Susceptiblity to bicarbonate-induced chlorosis was inversely correlated with both the Fe content in young leaves and the reducing capacity of roots, but not with the phosphorus content in young leaves. The plum Puebla de Soto 101 and the hybrid GF677 showed the lowest degree of chlorosis and the highest reducing capacity. Phosphate did not induce chlorosis.     

Genetic analysis of iron chlorosis tolerance in Prunus rootstocks

The high economic losses caused by the occurrence of iron chlorosis in Prunus orchards in the Mediterranean area justifies the implementation of breeding programs to generate high-performance rootstocks for different edaphoclimatic area conditions. For that reason, the genetic control of iron chlorosis tolerance was studied in an F₁ population derived from a three-way interspecific cross between a Myrobalan plum (P 2175) and an almond?×?peach hybrid (Felinem). Several phenotypic measurements were assessed to guarantee an accurate data set for genetic analysis. SPAD (Soil and Plant Analyzer Development) values, chlorophyll concentration, and visual diagnostic symptoms were highly correlated with leaf chlorosis in trees. SPAD value was the most reliable measure, since it was an objective, unbiased, and non-destructive method. Two significant quantitative trait loci (QTLs) involved in SPAD and chlorophyll concentration were identified for Felinem in linkage groups 4 and 6. Both QTLs were detected in four of the six consecutive years of the experiment. For P 2175, two of the three putative QTLs identified, pspad4.1 and chl4.1, were placed in linkage group 4. These QTLs were related to the SPAD values and chlorophyll concentration, respectively, and co-localized with QTLs detected in the Felinem map affecting the same traits. Candidate gene PFIT, related to iron metabolism, was localized within the confidence interval of the QTL in linkage group 4. This research suggests an association of this chromosome region with tolerance to iron chlorosis in Prunus, and it provides a first approach to localize candidate genes involved in tolerance to this abiotic stress.     

Responses of two Prunus rootstocks to KCl induced salinity in vitro

The in vitro response of two Prunus rootstocks: GF 677 (Prunus persica × Prunus amygdalus), and Nemared (Prunus persica) to increasing concentrations of KCl of the culture medium was studied. Shoots were grown in vitro for 8 weeks on an Murashige and Skoog medium supplemented with 0, 5, 10, 15, 20, 40 or 80 mM KCl. By increasing KCl concentration from 0 to 40 mM, the number of shoots per explant was not significantly affected for both rootstocks. However, Nemared rootstock formed more shoots per explant than GF 677 under respective KCl concentrations of the medium. Inclusion of 80 mM KCl in the medium resulted in a reduction of growth of both rootstocks. Sodium, Fe, Mn, and Zn concentration in tissues of Nemared rootstock were significantly higher than the respective values of GF 677.     

Leaf structural changes associated with iron deficiency chlorosis in field-grown pear and peach: physiological implications

Plants grown in calcareous, high pH soils develop Fe deficiency chlorosis. While the physiological parameters of Fe-deficient leaves have been often investigated, there is a lack of information regarding structural leaf changes associated with such abiotic stress. Iron-sufficient and Fe-deficient pear and peach leaves have been studied, and differences concerning leaf epidermal and internal structure were found. Iron deficiency caused differences in the aspect of the leaf surface, which appeared less smooth in Fe-deficient than in Fe-sufficient leaves. Iron deficiency reduced the amount of soluble cuticular lipids in peach leaves, whereas it reduced the weight of the abaxial cuticle in pear leaves. In both plant species, epidermal cells were enlarged as compared to healthy leaves, whereas the size of guard cells was reduced. In chlorotic leaves, bundle sheaths were enlarged and appeared disorganized, while the mesophyll was more compacted and less porous than in green leaves. In contrast to healthy leaves, chlorotic leaves of both species showed a significant transient opening of stomata after leaf abscission (Iwanoff effect), which can be ascribed to changes found in epidermal and guard cells. Results indicate that Fe-deficiency may alter the barrier properties of the leaf surface, which can significantly affect leaf water relations, solute permeability and pest and disease resistance.     

Floral analysis as a tool to diagnose iron chlorosis in orange trees

A three-year field experiment was conducted in a commercial orange grove [Citrus sinensis (L.) Osb. cv. `Valencia late' grafted on Citrange Troyer] established on a calcareous soil in the south of Portugal, to investigate if flower analysis could be used to diagnose lime-induced iron chlorosis. In April, during full bloom, flowers and leaves were collected from 20 trees. Leaf samples were again collected from the same trees in May, June, July and August. Total chlorophyll was estimated in all the leaves sampled for foliar analysis, using a SPAD-502 apparatus. Leaves and flowers were analysed for N, P, K, Ca, Mg, Fe, Zn, Mn and Cu. Principal Component Analysis was used to evaluate the variation of nutrient concentrations in flowers, and linear regressions were established between these and the chlorophyll content of leaves 90 days after full bloom. Evaluation of the best-fit equation was carried out using separate data obtained from other groves. Variation in the pattern of floral mineral composition in the flowers showed contrasts between the increase in N, P and K and that of Ca, Fe and Zn, while the concentration of Mg, Mn and Ca varied synchronously. The ratio of Mg:Zn in flowers explained about half of the variation of chlorophyll in leaves later in the season. A ratio below 100 indicated that trees would develop iron chlorosis, while with a ratio above 200 leaves would remain green. An early prognosis of iron chlorosis based on floral analysis can benefit growers, since it allows them to apply treatments in time to prevent loss of fruit yield and quality due to iron chlorosis.     

Mapping genetic loci for tolerance to lime-induced iron deficiency chlorosis in grapevine rootstocks (Vitis sp.)

Iron is essential to plants for chlorophyll formation as well as for the functioning of various iron-containing enzymes. Iron deficiency chlorosis is a wide-spread disorder of plants, in particular, of those growing on calcareous soils. Among the different ways to control iron deficiency problems for crops, plant material and especially rootstock breeding is a suitable and reliable method, especially for fruit trees and grapes. The aim of the experiment was to characterize the genetic basis of grapevine chlorosis tolerance under lime stress conditions. A segregating population of 138 F1 genotypes issued from an inter-specific cross between Vitis vinifera Cabernet Sauvignon (tolerant) × V. riparia Gloire de Montpellier (sensitive) was developed and phenotyped both as cuttings and as rootstock grafted with Cabernet Sauvignon scions in pots containing non-chlorosing and chlorosing soils. Tolerance was evaluated by chlorosis score, leaf chlorophyll content and growth parameters of the shoots and roots. The experiments were performed in 2001, 2003 and 2006. The plants analysed in 2006 were reassessed in 2007. The most significant findings of the trial were: (a) the soil properties strongly affect plant development, (b) there are differences in tolerance among segregating genotypes when grown as cuttings or as rootstocks on calcareous soil, (c) calcareous conditions induced chlorosis and revealed quantitative trait loci (QTLs) implicated in polygenic control of tolerance, (d) rootstock strongly contributes to lime-induced chlorosis response, and (e) a QTL with strong effect (from 10 to 25 % of the chlorotic symptom variance) was identified on chromosome 13. This QTL colocalized with a QTL for chlorophyll content (R ² = 22 %) and a major QTL for plant development that explains about 50 % of both aerial and root system biomass variation. These findings were supported by stable results among the different years of experiment. These results open new insights into the genetic control of chlorosis tolerance and could aid the development of iron chlorosis-tolerant rootstocks.     

Physiological and biochemical responses of the iron chlorosis tolerant grapevine rootstock 140 Ruggeri to iron deficiency and bicarbonate

Background and aims

Iron (Fe) deficiency chlorosis associated with high levels of soil bicarbonate is one of the main nutritional disorders observed in sensitive grapevine genotypes. The aim of the experiment was to assess both the independent and combined effects of Fe and bicarbonate nutrition in grapevine.

Methods

Plants of the Fe chlorosis tolerant 140 Ruggeri rootstock were grown with and without Fe(III)-EDTA and bicarbonate in the nutrient solution. SPAD index, plant growth, root enzyme (PEPC, MDH, CS, NADP⁺ ?IDH) activities, kinetic properties of root PEPC, organic acid concentrations in roots and xylem sap and xylem sap pH were determined. A factorial statistical design with two factors (Fe and BIC) and two levels of each factor was adopted: +Fe and ?Fe, and +BIC and ?BIC.

Results

This rootstock strongly reacted to Fe deficiency by activating several response mechanisms at different physiological levels. The presence of bicarbonate in the nutrient solution changed the activity of PEPC and TCA related enzymes (CS, NADP⁺-IDH) and the accumulation/translocation of organic acids in roots of Fe-deprived plants. Moreover, this genotype increased root biomass and root malic acid concentration in response to high bicarbonate levels in the substrate. Bicarbonate also enhanced leaf chlorophyll content.

Conclusions

Along with a clear independent effect on Fe nutrition, our data support a modulating role of bicarbonate on Fe deficiency response mechanisms at root level.     

Manganese Toxicity in Sugarcane Plantlets Grown on Acidic Soils of Southern China

Ratoon sugarcane plantlets in southern China have suffered a serious chlorosis problem in recent years. To reveal the causes of chlorosis, plant nutrition in chlorotic sugarcane plantlets and the role of manganese (Mn) in this condition were investigated. The study results showed that the pH of soils growing chlorotic plantlets ranged from 3.74 to 4.84. The symptoms of chlorosis were similar to those of iron (Fe) deficiency while the chlorotic and non-chlorotic plantlets contained similar amount of Fe. Chlorotic plantlets had 6.4-times more Mn in their leaf tissues compared to the control plants. There was a significantly positive correlation between Mn concentration in the leaves and the exchangeable Mn concentration in the soils. Moreover, leaf Mn concentration was related to both seasonal changes in leaf chlorophyll concentration and to the occurrence of chlorosis. Basal stalks of mature sugarcanes contained up to 564.36 mg·kg^-1 DW Mn. Excess Mn in the parent stalks resulted in a depress of chlorophyll concentration in the leaves of sugarcanes as indicated by lower chlorophyll concentration in the leaves of plantlets emerged from basal stalks. Ratoon sugarcane plantlets were susceptible to chlorosis due to high Mn accumulation in their leaves (456.90–1626.95 mg·kg^-1 DW), while in planted canes chlorosis did not occur because of low Mn accumulation (94.64–313.41mg·kg^-1 DW). On the other hand, active Fe content in chlorotic plantlets (3.39 mg kg^-1 FW) was only equivalent to 28.2% of the concentration found in the control. These results indicate that chlorosis in ratoon sugarcane plantlets results from excessive Mn accumulated in parent stalks of planted cane sugarcanes grown on excessive Mn acidic soils, while active Fe deficiency in plantlets may play a secondary role in the chlorosis.     

Influence of rootstock on antioxidant system in leaves and roots of young apple trees in response to drought stress

Grafting rootstocks are widely used to enhance plants resistance to various biologic and abiotic stresses. We determined how the rootstock genotype might influence plant responses to drought, using 2-year-old ‘Gale Gala’ apple trees grafted onto Malus sieversii and M. hupehensis. Under water stress, trees with the former as their rootstock had smaller reductions in rates of relative growth and photosynthesis, total biomass, leaf area, levels of leaf chlorophyll, and relative water content compared with those grafted onto the latter. They also had greater maximum photochemical efficiency and water-use efficiency. On the other hand, trees growing on M. sieversii rootstock had less production of superoxide radicals and hydrogen peroxide in both leaves and roots than those growing on M. hupehensis in response to drought stress. Furthermore, under drought conditions, leaves and roots from trees grafted onto M. sieversii had greater synthesis of ascorbic acid and glutathione, as well as higher activities of superoxide dismutase, catalase, ascorbate peroxidase, monodehydroascorbate reductase, dehydroascorbate reductase, and glutathione reductase. These results suggest that the choice of grafting rootstock can enhance drought resistance by improving the antioxidant system in a plant. Here, ‘Gale Gala’ trees grafted onto M. sieversii were more drought-resistant than those on M. hupehensis rootstock.     

An Exogenous Source of Nitric Oxide Modulates Iron Nutritional Status in Peanut Seedlings (<Emphasis Type="Italic">Arachis hypogaea</Emphasis> L.)

Iron (Fe) deficiency chlorosis is a common and severe nutritional deficiency in plants, and nitric oxide (NO) is an important signaling molecule in regulating Fe homeostasis in plants. We studied the effect of sodium nitroprusside (SNP, an NO donor) on Fe uptake, translocation, storage, and activation in a greenhouse. The concentrations of active Fe, total Fe, and the ratio of active Fe to total Fe, the activities of key enzymes, and chlorophyll concentration were determined, and resistance to oxidative stress and mineral element distribution in peanut plants grown in Fe sufficiency and Fe deficiency (an absence of Fe and low level of Fe concentration) conditions were also investigated. The results showed that NO significantly increased the concentration of active Fe and the ratio of active Fe to total Fe in Fe-deficient plants, and increased active Fe concentration in leaves and stems of Fe-sufficient plants. NO application also increased Fe translocation from roots to the shoots and the accumulation of Fe in cell organelles and the soluble fraction in leaves, especially in the low-level Fe concentration condition, thus increased available Fe and chlorophyll concentration in leaves of Fe-deficient plants. The activities of key enzymes were regulated by NO, which effectively mitigated oxidative damages by enhancing the activities of antioxidant enzymes (SOD, POD, CAT), increasing H⁺-ATPase and Ca²⁺-ATPase activities to balance the ion (Fe, Ca, Mg and Zn) uptake and distribution in Fe-deficient plants. However, NO application had no obvious effect on these variables in Fe-sufficient plants. These results indicated that NO application can improve Fe uptake, translocation, and activation of related enzymes in Fe-deficient plants, thus mitigating the adverse effect of Fe deficiency.     

Chlorosis correction and agronomic biofortification in field peas through foliar application of iron fertilizers under Fe deficiency

Effectiveness of different iron (Fe) foliar sprays for leaf chlorosis correction and grain Fe boosting was studied in field peas under Fe deficiency. No chlorophyll reduction was observed in Fe deficient plants treated with foliar sprays. EDDHA [ethylenediamine-N,N′-bis(2-hydroxyphenylacetic acid)] followed by FeSO₄ (73.7?mg/l Fe) treated at the start of flowering was most responsive in correcting chlorosis and increasing shoot dry biomass in peas. Inductively coupled plasma-atomic emission spectroscopy data showed significant increase of Fe in grains while treated with all foliar sprays at the time of grain filling in Fe-deficient plants. Among them, FeSO₄ (73.7?mg/l Fe) was the most efficient in biofortifying Fe in mature grain under Fe deficiency in peas. Results also pinpoint that flowering is a suitable time for applying foliar sprays to boost Fe in mature grains. Taken together, application of Fe foliar sprays facilitated both chlorosis correction and Fe boosting in peas and can be further used by breeders and farmers.     

Iron deficiency-associated changes in the composition of the leaf apoplastic fluid from field-grown pear (Pyrus communis L.) trees.

Experiments have been carried out with field-grown pear trees to investigate the effect of iron chlorosis on the composition of the leaf apoplast. Iron deficiency was associated with an increase in the leaf apoplastic pH from the control values of 5.5-5.9 to 6.5-6.6, as judged from direct pH measurements in apoplastic fluid obtained by centrifugation and fluorescence of leaves incubated with 5-CF. The major organic acids found in leaf apoplastic fluid of iron-deficient and iron-sufficient pear leaves were malate, citrate and ascorbate. The total concentration of organic acids was 2.9 mM in the controls and increased to 5.5 mM in Fe-deficient leaves. The total apoplastic concentration of inorganic cations (Ca, K and Mg) increased with Fe deficiency from 15 to 20 mM. The total apoplastic concentration of inorganic anions (Cl-, NO3-, SO4(2-) and HPO4(2-)) did not change with Fe deficiency. Iron concentrations decreased from 4 to 1.6 microM with Fe deficiency. The major Fe species predicted to exist in the apoplast was [FeCitOH](-1) in both Fe-sufficient and deficient leaves. Organic acids in whole leaf homogenates increased from 20 to 40 nmol x m(-2) with Fe deficiency. The accumulation of organic anions in the Fe-deficient leaves does not appear to be associated to an increased C fixation in leaves, but rather it seems to be a consequence of C transport via xylem.     

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.     

Water-extractable humic substances enhance iron deficiency responses by Fe-deficient cucumber plants

The ability of Fe-deficient cucumber plants to use iron complexed to a water-extractable humic substances fraction (WEHS), was investigated. Seven-day-old Fe-deficient plants were transferred to a nutrient solution supplemented daily for 5 days with 0.2 μM Fe as Fe-WEHS (5 μg org. C mL^-1), Fe-EDTA, Fe-citrate or FeCl₃. These treatments all allowed re-greening of the leaf tissue, and partial recovery of dry matter accumulation, chlorophyll and iron contents. However, the recovery was faster in plants supplied with Fe-WEHS and was already evident 48 h after Fe supply. The addition of 0.2 μM Fe to the nutrient solution caused also a partial recovery of the dry matter and iron accumulation in roots of Fe-deficient cucumber plants, particularly in those supplied with Fe-WEHS. The addition of WEHS alone (5 μg org. C mL^-1, 0.04 μM Fe) to the nutrient solution slightly but significantly increased iron and chlorophyll contents in leaves of Fe-deficient plants; in these plants, dry matter accumulation in leaves and roots was comparable or even higher than that measured in plants treated with Fe-citrate or FeCl₃. After addition of the different iron sources for 5 days to Fe-deficient roots, morphological modifications (proliferation of lateral roots, increase in the diameter of the sub-apical zones and amplified root-hair formation) and physiological responses (enhanced Fe(III)-chelate reductase and acidification of the nutrient solution) induced by Fe deficiency, were still evident, particularly in plants treated with the humic molecules. The presence of WEHS caused also a further acidification of the nutrient medium by Fe-deficient plants. The Fe-WEHS complex (1 μM Fe) could be reduced by intact cucumber roots, at rates of reduction higher than those measured for Fe-EDTA at equimolar iron concentration. Plasma membrane vesicles, purified by two-phase partition from root microsomes of Fe-deficient plants, were also able to reduce Fe-WEHS. Results show that Fe-deficient cucumber plants can use iron complexed to water soluble humic substances, at least in part via reduction of complexed Fe(III) by the plasma membrane Fe(III)-chelate reductase of root cells. In addition, the stimulating effect of humic substances on H⁺ release might be of relevance for the overall response of the plants to iron shortage. This revised version was published online in June 2006 with corrections to the Cover Date.     

Physiological responses of Tunisian grapevine varieties to bicarbonate-induced iron deficiency

Plants are frequently submitted to iron deficiency when growing on calcareous soils, which contain high concentrations of bicarbonate. The purpose of this study was to investigate the variability of physiological responses of Tunisian grapevine varieties to bicarbonate-induced iron chlorosis. Vine woodcuttings of seven autochthonous Tunisian varieties (Khamri, Mahdaoui, Blan3, Saouadi, Arich Dressé, Beldi and Balta4), two rootstocks (140Ru and S.O.4), and an introduced table variety (Cardinal) were cultivated on inert sand for 2 months using a complete nutrient solution (20 microM Fe) that was either well supplied or not supplied with 10 mM HCO3-. Young leaves of plants cultivated on bicarbonate-enriched medium showed characteristic symptoms of iron chlorosis, although the intensity of the symptoms depended on the variety and the rootstock. Chlorosis score confirmed these observations since the most sensitive varieties showed the highest values. This variability in tolerance to iron deficiency was also displayed when analysing the physiological parameters (shoot length, plant dry weight, and chlorophyll concentration) and the iron contents in the 4th leaf. Analysis of morphological and physiological parameters showed three behaviour groups. The first one corresponded to tolerant varieties (Khamri, Mahdaoui, and the root-stock: 140Ru), the second included moderately tolerant vines (Saouadi, Arich Dressé, Blanc3, and the rootstock: S.O.4) and the third represented the sensitive ones (Balta4, Beldi, and Cardinal).     

Effects of Exogenous Salicylic Acid on Alleviating Chlorosis Induced by Iron Deficiency in Peanut Seedlings (Arachis hypogaea L.)

The effects of salicylic acid (SA) on alleviating chlorosis induced by iron (Fe) deficiency in peanut seedlings (Arachis hypogaea L.) were studied by investigating the symptoms, plant growth, chlorophyll concentrations, soluble Fe concentration, Fe distribution in subcellular, and antioxidant enzymes. Fe deficiency caused serious chlorosis and inhibited growth of peanut seedlings, and dramatically decreased the soluble Fe concentration and chlorophyll concentration. Furthermore, ion balance was disturbed. The addition of 50, 100, and 250 μM SA significantly increased the absorption of Fe from the cell wall to cell organelles and the soluble fraction, enhanced the Fe concentration in cell organelles, Fe activation and chlorophyll concentrations in leaves, ameliorated the inhibition of Ca, Mg, and Zn absorption induced by Fe deficiency, alleviated the chlorosis induced by Fe deficiency and promoted plant growth. The accumulation of reactive oxygen species (ROS) is dramatically increased in peanut seedlings exposed to Fe deficiency, and resulted in lipid peroxidation, which was indicated by accumulation of malondialdehyde (MDA). The application of 50, 100, and 250 μM SA significantly decreased the level of ROS and MDA concentrations, and significantly increased the activities of superoxide dismutase, peroxidase, and catalase in peanut seedlings exposed to Fe deficiency. The addition of 100 μM SA had the best effect on alleviating chlorosis induced by Fe deficiency, whereas the addition of 500 μM SA had no significant effect under Fe deficiency.     

Iron Enhances Aluminum-induced Leaf Necrosis and Plant Growth Inhibition in Eucalyptus camaldulensis

The combined effects of excess Fe and Al on Eucalyptus camaldulensis Dehnh. were studied by investigating time course and visible symptoms of leaf necrosis, plant biomass, the status of some antioxidants and pigments and nutrient concentrations. Seedlings were grown hydroponically in nutrient solutions containing 0 or 500 μM AlCl₃, each with a FeSO₄ range of 1, 12 and 120 μM at pH 4.2. Leaf necrosis and plant growth inhibition were induced by Al and enhanced by the increase in Fe concentration. The process from the first appearance of necrotic spots to leaf death (shedding) of a leaf proceeded from a few days to about 20 days after the leaf had fully expanded. Either 120 μM Fe without Al or Al reduced plant growth to a similar extent but 120 μM Fe without Al did not cause leaf necrosis. In leaves, excess Fe (12 and 120 μM) without Al reduced concentration of ASC and GSH, while concentration of Fe, DHA and GSSG and DHA:ASC and GSSG:GSH ratios tended to increase with the increase in Fe concentration in treatment solution with or without Al. At 1 μM Fe, Al increased concentration of DHA and DHA:ASC and GSSG:GSH ratios. Catalase activity in leaves reduced with the increase in leaf Al concentration. At 1 μM Fe, Al greatly reduced concentrations of Fe and chlorophylls in leaves but increase two times Fe concentration in stems. These suggest that the enhancement effects of Fe on Al-induced leaf necrosis and plant growth inhibition can be discussed in context of the excess Fe itself weakens antioxidant capability of ASC–GSH cycle in leaves and greatly reduces plant growth; and the increase in Fe accumulation in stems is involved in Al-induced leaf chlorosis.