Selection of olive varieties for tolerance to iron chlorosis

Under certain conditions, olive trees grown on calcareous soils suffer from iron chlorosis. In the present study several olive varieties and scion-rootstock combinations were evaluated for their tolerance to iron chlorosis. Plants were grown over several months in pots with a calcareous soil, under two fertilization treatments. These consisted of periodic applications of nutrient solutions containing either, 30 μmol/L FeEDDHA or not Fe. Tolerance was assessed by the chlorosis and growth parameters of plants grown without Fe, compared to those plants grown with Fe. Results show that there are differences in tolerance among olive varieties and that tolerance is mainly determined by the genotype of the rootstock. These results open the way to use tolerant varieties for those conditions where iron chlorosis could become a problem.     

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.     

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.     

Differences in responses to iron deficiency among various cultivars of mungbean (Vigna radiata (L.) Wilczek)

Ten mungbean cultivars were evaluated for their resistance to iron deficiency in view of chlorosis symptoms, plant growth and seed yield under field conditions on a calcareous soil in Thailand. The KPS2 cultivar was highly susceptible; the KPS1, PSU1 and Pag-asa 1 cultivars were somewhat susceptible; the VC1163B cultivar was moderately tolerant; the CN36, CN60, UT1 and CNM-I cultivars were tolerant; and the CNM8509B cultivar was very tolerant to iron deficiency. Foliar application of a solution of 5 g L^-1 ferrous sulphate was effective in correcting chlorosis that was induced by iron deficiency, and it enhanced both the growth and the yield of susceptible cultivars. Compared with the susceptible cultivar KPS2, the tolerant cultivar UT1 had a greater ability to lower the pH of the nutrient solution in response to iron deficiency. The root-associated Fe³⁺-reduction activity of UT1 that had been grown in -Fe medium was similar to that of the plants grown in +Fe medium when the acidification of the medium occurred. Acidification of the medium in response to iron deficiency might contribute to the efficient solubilization of iron from calcareous soils, and it related more closely to the resistance to iron deficiency than Fe³⁺ reduction by roots in mungbean cultivars.     

Overexpression of AtFRO6 in transgenic tobacco enhances ferric chelate reductase activity in leaves and increases tolerance to iron-deficiency chlorosis

The Arabidopsis gene FRO6(AtFRO6) encodes ferric chelate reductase and highly expressed in green tissues of plants. We have expressed the gene AtFRO6 under the control of a 35S promoter in transgenic tobacco plants. High-level expression of AtFRO6 in transgenic plants was confirmed by northern blot analysis. Ferric reductase activity in leaves of transgenic plants grown under iron-sufficient or iron-deficient conditions is 2.13 and 1.26 fold higher than in control plants respectively. The enhanced ferric reductase activity led to increased concentrations of ferrous iron and chlorophyll, and reduced the iron deficiency chlorosis in the transgenic plants, compared to the control plants. In roots, the concentration of ferrous iron and ferric reductase activity were not significantly different in the transgenic plants compared to the control plants. These results suggest that FRO6 functions as a ferric chelate reductase for iron uptake by leaf cells, and overexpression of AtFRO6 in transgenic plants can reduce iron deficiency chlorosis.     

Humic acid and iron uptake by plants

Summary The behaviour of ferric EDTA and ferric citrate in nutrient solution and their interaction with humic acid was investigated at various hydrogen ion concentrations using the technique of membrane ultrafiltration to separate small iron species from high molecular weight products of hydrolysis and to estimate the binding of iron by humic acid. Ferric EDTA was found to be of small molecular size at all pH values between 5.0 and 7.0 whilst ferric citrate solutions contained an increasing proportion of high molecular weight material as pH was increased from 5.0 to 7.0. Some iron present in solutions of both ferric EDTA and ferric citrate was bound by humic acid at all pH values from 5.0 to 7.0. Studies were also made of the uptake of iron by wheat roots from nutrient solutions containing either ferric EDTA or ferric citrate and of the effect of humic acid on uptake. More iron was absorbed from ferric EDTA than from ferric citrate at all pH values. Increasing pH between 5.0 and 7.0 resulted in a progressive decrease in the uptake of iron in both cases. The presence of humic acid depressed iron absorption from both solutions at all pH values.     

On the effect of ferrous sulphate on the available manganese in the soil and the uptake of manganese by the plant

Summary Application of iron as ferrous sulphate or chloride to a loam not deficient in manganese had no effect on the yield but increased the uptake of manganese even in barley which grew vigorously.In an experiment with sugar beet on two soils contrasted with regard to their available manganese supply, applications of manganese, iron and nitrogen were tested in all combinations. On the Købelev soil, not deficient in manganese, no increases in yields were obtained on addition of ferrous sulphate while increases in manganese uptake were found for all combinations of treatments except where iron was added in the presence of manganese.On the manganese deficient Faarevejle soil, significantly higher increases in yields of roots were obtained from ferrous sulphate in the presence of nitrogen than in the presence of manganese. The effect of iron in the presence of nitrogen on the yield of tops was also significant. These treatments also gave the highest increases in manganese uptake.The amounts of manganese extractable from the soils by magnesium nitrate over a range of pH 2–8 could be increased considerably by addition of ferrous sulphate.The results support the suggestion that application of ferrous sulphate to some soils has the same effect as an addition of manganese.     

Effect of soil and foliar application of ferrous sulphate and of acidulation of soil on iron chlorosis of paddy seedlings ingoradu soil nurseries in India

Summary Nursery experiments were conducted to determine the cause for the chronic problem of chlorosis in paddy seedlings raised ongoradu soil nurseries of Anand area of Kheda district of Gujarat State and to find out remedial measures for it. The findings are (i) the chlorosis is due to high bicarbonate content of the soil (ii) application of ferrous sulphate to soil at 40 kg Fe/ha increased the survival period of the seedling but did not quite control the chlorosis (iii) spray of 0.4% ferrous sulphate solution twice a week was helpful in partial recovery of the seedlings from chlorosis (iv) acidulation of soil with sulphuric acid about ten days prior to sowing produced very healthy, vigorous and green seedlings of paddy which did not show any signs of chlorosis at all and (v) total and IN HCl-soluble iron content of the shoots did not at all reflect the degree of chlorosis of the paddy seedlings. These results are discussed in the light of current theories of absorption and translocation of iron in plant system and the iron-chlorosis of paddy seedlings ingoradu soils is attributed to both soil and plant factors.     

黄瓜叶片对草酸铁的还原作用

铁还原作用在植物叶片对铁素吸收及利用过程中起关键作用.本研究表明相对于其它几种常用的铁螯合物如二乙基四乙酸铁(FeⅢEDTA)或柠檬酸铁,草酸铁更有利于黄瓜活体叶片及铁还原酶的作用,即表现出更高的铁还原活力.缺铁降低了黄瓜叶片中的铁还原活性.缺铁时叶片中的草酸含量不受影响,而富含在石灰性缺铁土壤中的碳酸氢根离子能使叶片中草酸含量显著提高.     

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

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

Photosynthetic carbon fixation in iron-chlorotic and recovered green sugarcane leaves

Summary Sugarcane var. Co 740 is grown in various parts of Maharashtra (India) and is susceptible to chlorosis due to physiological non-utilization of iron. The physiological disorder is seen over a large area and it results in poor yield. Low sucrose yield can be recovered by foliar sprays of ferrous sulphate. The nonchlorotic and chlorotic leaves were used for the photosynthetic studies. The leaves after ferrous sulphate treatment show an increase in total chlorophyll contents and at the same time show an improved chlorophylla to chlorophyllb ratio which is affected in the chlorotic ones. The recovered green leaves have higher uptake of nitrogen, phosphorus, potassium and iron.¹⁴CO₂ fixation studies for short- and long-term experiments reveal that recovered green leaves can synthesize malate more efficiently and also utilize it for sucrose synthesis more rapidly than in the chlorotic ones. On the contrary more amino acids, reducing sugars and sugar phosphates are synthesized in the chlorotic leaves. There is also an accumulation of citrate, glutamate, and tartrate in the chlorotic leaves. Our results indicate that sucrose synthesis is disturbed in the chlorotic leaves and can be corected byfolia: sprays of ferrous sulphate.     

Association of cation and organic anion accumulation with iron chlorosis of Scots pine on prairie soils

Summary Iron chlorosis of 4 year old Scots pine (Pinus sylvestris L.) in comparison to areas of adjacent healthy growth on calcareous prairie soil, was associated with slight increases in the soluble ion content of the saturation paste extract. Such increases in soluble ions (mainly calcium sulphate) were associated with significant increases in ash, cation (including iron) and organic anion content of the chlorotic needles. Increasing levels of available soil nitrate were also related to increase in organic anions. Nitrogen and phosphorus assimilation was adversely affected under conditions of iron chlorosis. These observations support the theory of induced iron deficiency associated with elevated levels of organic anions or translocated cations and are applicable to plantings of conifers on prairie soils.     

Functional Analysis of the Genotypic Differences in Response of Pea (<i>Pisum sativum</i> L.) to Calcareous-Induced Iron Deficiency

Lime-induced iron chlorosis is a major nutritional disorder causing severe plant growth and yield reduction in the calcareous soils of Tunisia. The understanding the behavior of key metabolic functions of peas on calcareous soils, the identification of useful traits of tolerance, and the exploration of the genotypic differences in response to this constraint remain the most efficient approaches due to their coast, environmental benefits, and sustainability. For this purpose, a greenhouse experiment was conducted on three pea genotypes (Alexandra: Alex, Douce de provence: DP, and Merveille de Kelvedon: MK) cultivated on calcareous soil (Fe-deficient) and fertile soil (control). Plant growth, SPAD index, iron nutrition and distribution, photosynthesis, and antioxidant enzymes were deeply analyzed to discriminate genotypic differences. Calcareous-induced iron deficiency reduced SPAD index, plant growth, net photosynthesis, and tissue Fe content against a significant stimulation of the oxidative stress indicators, H₂O₂ and Malondialdehyde (MDA). Moreover, we reported a significant induction of SOD and CAT activity in shoots and roots of the Alexandra genotype. Fe use efficiency increased on calcareous soil and clearly discriminated the studied genotypes. Alexandra genotype was found to be the most tolerant to lime-induced iron chlorosis. This genotype protects its tissues against oxidative stress by stimulating enzyme activities (SOD and CAT) and develops significant efficiency of Fe uptake, translocation to shoots and use when cultivated on calcareous soil.     

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

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

Iron and gallium increase iron uptake from transferrin by human melanoma cells: further examination of the ferric ammonium citrate-activated iron uptake process

Previously we showed that preincubation of cells with ferric ammonium citrate (FAC) resulted in a marked increase in Fe uptake from both (59)Fe-transferrin (Tf) and (59)Fe-citrate (D.R. Richardson, E. Baker, J. Biol. Chem. 267 (1992) 13972-13979; D.R. Richardson, P. Ponka, Biochim. Biophys. Acta 1269 (1995) 105-114). This Fe uptake process was independent of the transferrin receptor and appeared to be activated by free radicals generated via the iron-catalysed Haber-Weiss reaction. To further understand this process, the present investigation was performed. In these experiments, cells were preincubated for 3 h at 37 degrees C with FAC or metal ion solutions and then labelled for 3 h at 37 degrees C with (59)Fe-Tf. Exposure of cells to FAC resulted in Fe uptake from (59)Fe-citrate that became saturated at an Fe concentration of 2.5 microM, while FAC-activated Fe uptake from Tf was not saturable up to 25 microM. In addition, the extent of FAC-activated Fe uptake from citrate was far greater than that from Tf. These results suggest a mechanism where FAC-activated Fe uptake from citrate may result from direct interaction with the transporter, while Fe uptake from Tf appears indirect and less efficient. Preincubation of cells with FAC at 4 degrees C instead of 37 degrees C prevented its effect at stimulating (59)Fe uptake from (59)Fe-Tf, suggesting that an active process was involved. Previous studies by others have shown that FAC can increase ferrireductase activity that may enhance (59)Fe uptake from (59)Fe-Tf. However, there was no difference in the ability of FAC-treated cells compared to controls to reduce ferricyanide to ferrocyanide, suggesting no change in oxidoreductase activity. To examine if activation of this Fe uptake mechanism could occur by incubation with a range of metal ions, cells were preincubated with either FAC, ferric chloride, ferrous sulphate, ferrous ammonium sulphate, gallium nitrate, copper chloride, zinc chloride, or cobalt chloride. Stimulation of (59)Fe uptake from Tf was shown (in order of potency) with ferric chloride, ferrous sulphate, ferrous ammonium sulphate, and gallium nitrate. The other metal ions examined decreased (59)Fe uptake from Tf. The fact that redox-active Cu(II) ion did not stimulate Fe uptake while redox-inactive Ga(III) did, suggests a mechanism of transporter activation not solely dependent on free radical generation. Indeed, the activation of Fe uptake appears dependent on the presence of the Fe atom itself or a metal ion with atomic similarities to Fe (e.g. Ga).     

Effect of salt on physiological responses of barley to iron deficiency.

Iron chlorosis is very common on alkaline soils such as calcareous ones, since iron availability is limited by high pH. Under these conditions of iron deficiency, graminaceous plant species induce special mechanisms for iron acquisition, involving enhanced release of iron chelators called phytosiderophores. On the other hand, it is known that most of salt soils have alkaline pH. So, plants growing on this kind of soils are often subjected simultaneously to salinity and iron deficiency. This work aimed at (i) studying the physiological responses of barley (Hordeum vulgare L.) to iron deficiency, and (ii) evaluating the effect of salt on the iron nutrition and the phytosiderophore release. For this purpose, seedlings of Hordeum vulgare L. were cultivated under controlled conditions, either in a complete nutrient solution with or without NaCl, or in an iron free nutrient solution containing or not NaCl. The plant morphological aspect, chlorophyll content of young leaves, iron status, biomass production, and phytosiderophore release by roots were assessed. Plants subjected to Fe deficiency exhibited a severe chlorosis, accompanied by a significant biomass reduction. These plants developed more lateral roots than the control with a highly stimulated phytosiderophore release. However, the latter was greatly diminished when iron deficiency was associated to salinity. A depressive effect of salt on iron acquisition in plants subjected only to salt stress which was also observed and further confirmed by the important decrease of efficiency in iron acquisition. These results suggest that salinity may reduce capacity of plants to acquire iron from alkaline soils by inhibiting phytosiderophore release.     

Apoplastic pH and FeIII reduction in young sunflower (Helianthus annuus) roots

The relationship between the apoplastic pH in young sunflower roots ( Helianthus annuus L.) and the plasmalemma ferric chelate reductase (FC-R; EC 1.16.1.7) activity in roots was investigated. The hypothesis was tested that a high apoplastic pH depresses FC-R activity, thereby restricting the uptake of Fe²⁺ into the cytosol. Until recently, little has been known about this relationship, because pH and redox reaction measurements are difficult to perform within the confines of the root apoplast. We recorded the apoplastic pH by means of the fluorescence ratio in conjunction with video microscopy by covalently tagging fluorescein boronic acid to OH groups of the root cell wall. Fe^III reduction was measured using a similar approach by tagging ferrozine diboronic acid with OH groups of the cell wall. Ferrozine forms an Fe²⁺ complex, thus indicating the reduction of ferric iron. In roots bathing in buffered outer solutions of different pH, a high pH sensitivity of apoplastic Fe^III reduction was found, with the highest ferric iron reduction rates at an apoplastic pH of 4.9; above an apoplastic pH of 5.3, no reduction was observed. Nitrate in the bathing solution increased the apoplastic pH and hence depressed the Fe^III reduction; ammonium had the reverse effect. Nitrate together with HCO₃^–, a combination which is typical of calcareous soils, had the strongest depressing effect. From the results, it can be concluded that the main reason for the frequently occurring iron deficiency chlorosis of plants grown on calcareous soils is the inhibition of Fe^III reduction in the apoplast, and hence Fe²⁺ uptake into the cytosol.     

Different iron sources to study the physiology and biochemistry of iron metabolism in marine micro-algae

We compared ferric EDTA, ferric citrate and ferrous ascorbate as iron sources to study iron metabolism in Ostreococcus tauri, Phaeodactlylum tricornutum and Emiliania huxleyi. Ferric EDTA was a better iron source than ferric citrate for growth and chlorophyll levels. Direct and indirect experiments showed that iron was much more available to the cells when provided as ferric citrate as compared to ferric EDTA. As a consequence, growth media with iron concentration in the range 1–100 nM were rapidly iron-depleted when ferric citrate—but not ferric EDTA was the iron source. When cultured together, P. tricornutum cells overgrew the two other species in iron-sufficient conditions, but E. huxleyi was able to compete other species in iron-deficient conditions, and when iron was provided as ferric citrate instead of ferric EDTA, which points out the critical influence of the chemical form of iron on the blooms of some phytoplankton species. The use of ferric citrate and ferrous ascorbate allowed us to unravel a kind of regulation of iron uptake that was dependent on the day/night cycles and to evidence independent uptake systems for ferrous and ferric iron, which can be regulated independently and be copper-dependent or independent. The same iron sources also allowed one to identify molecular components involved in iron uptake and storage in marine micro-algae. Characterizing the mechanisms of iron metabolism in the phytoplankton constitutes a big challenge; we show here that the use of iron sources more readily available to the cells than ferric EDTA is critical for this task.     

Bulk soil pH and rhizosphere pH of peach trees in calcareous and alkaline soils as affected by the form of nitrogen fertilizers

One-year old nectarine trees [Prunus persica, Batsch var. nectarina (Ait.) Maxim.], cv Nectaross grafted on P.S.B2 peach seedlings [Prunus persica (L.) Batsch] were grown for five months in 4-litre pots filled with two alkaline soils, one of which was also calcareous. Soils were regularly subjected to fertigation with either ammonium sulphate or calcium nitrate providing a total of 550 mg N/tree. Trees were also grown in such soils receiving only deionized water, as controls. Rhizosphere pH, measured by the use of a microelectrode inserted in agar sheet containing a bromocresol purple as pH indicator and placed on selected roots, was decreased by about 2–3 units compared to the bulk soil pH in all treatments. This decrease was slightly less marked when plants were supplied with calcium nitrate rather than ammonium sulphate or control. Measurements conducted during the course of the experiment indicated that ammonium concentration was similar in the solution of soils receiving the two N fertilizers. During the experiment, soil solution nitrate-N averaged 115 mg L^–1 in soil fertilized with calcium nitrate, 68 mg L^–1 in those receiving ammonium sulphate and 1 mg L^–1 in control soils. At the end of the experiment nitrate concentrations were similar in soils receiving the two N sources and bulk soil pH was decreased by about 0.4 units by ammonium sulphate fertigation: these evidences suggest a rapid soil nitriflcation activity of added ammonium. Symptoms of interveinal chlorosis in apical leaves appeared during the course of the experiment in trees planted in the alkaline-calcareous soil when calcium nitrate was added. The slightly higher rhizosphere pH for calcium nitrate-fed plants may have contributed to this. The findings suggest that using ammonium sulphate in a liquid form (e.g. by fertigation) in high-pH soils leads to their acidification and the micronutrient availability may be improved.