Effects of iron nutritional status and time of day on concentrations of phytosiderophores and nico‐tianamine in different root and shoot zones of barley

The diurnal pattern in concentrations of phytosiderophores (PS) and its precursor nicotianamine (NA) was studied in different root and shoot zones of iron (Fe)‐sufficient and Fe‐deficient barley (Hordeum vulgare L. cv. Europa) grown in nutrient solution. Roots were separated into apical (0–3 cm) and basal zones (>3 cm) and shoots into young (3 cm basal zones of youngest two leaves) and old (remaining zones of youngest two leaves and oldest leaf) parts. The main PS in barley was identified as epi‐hydroxymugineic acid (epi‐HMA). Regardless of the sampling zone and time of day, epi‐HMA concentrations were several times higher in Fe‐deficient than in Fe‐sufficient plants and several times higher in the roots than in the shoots. In roots and shoots, epi‐HMA concentrations were always higher in the younger compared with the older zones. In both root zones of Fe‐deficient plants, an inverse diurnal rhythm occurred in epi‐HMA concentrations and in its release by the roots. In contrast, such a rhythm was absent in roots of Fe‐sufficient plants and in the shoots regardless of the Fe nutritional status. Nicotianamine concentrations in roots were not affected by the Fe nutritional status in apical zones but slightly enhanced under Fe deficiency in basal zones. In contrast to roots, NA concentrations in both shoot parts were lower in Fe‐deficient than in Fe‐sufficient plants. Regardless of the Fe nutritional status in roots and shoots, NA concentrations were higher in young than in old parts and no consistent diurnal variations were observed. The results suggest that PS are also synthesized in the shoot, although at much lower rates than in roots. As with roots, PS synthesis in the shoot is enhanced under Fe deficiency and is mainly localized in young growing tissue. The distinct diurnal rhythm in PS release in roots is apparently not regulated by variation in the rate of PS synthesis during the day.     

Effectiveness of Phytosiderophore in Absorption and Translocation of 59Iron in Barley in the Presence of Plant-Borne,Synthetic, and Microbial Chelators

The mechanisms of iron (Fe) absorption and translocation in plants have received much study because they are the key processes in the supply of Fe to plants. The objective of this research was to study the effectiveness of phytosiderophore (PS) in the absorption and translocation of ⁵⁹Fe in Fe-deficient barley (Hordeum vulgare L. cv. ‘Minorimugi’) plants in the presence of plant-borne, synthetic, or microbial chelators. Plants grown under Fe-deficient conditions in a phytotron at pH 5.5 for 7–18 d were fed with Fe³⁺ (10 μ M labeled with ⁵⁹Fe) in the presence of 10 μ M of different chelators with or without 10 μ M PS for 4 h starting at 2 p.m. (6 h after the onset of light period). The absorption and translocation of ⁵⁹Fe in plants treated with PS and Fe^{3 +} were increased relative to plants fed solely with Fe^{3 +} (control). There was no effect found on absorption and translocation of ⁵⁹Fe in plants treated with EDTA or p-coumarate relative to the control, but a differential increase was observed in ⁵⁹Fe absorption and translocation in plants treated with EDTA or p-coumarate in the presence of PS. In comparison with the control, a decrease in ⁵⁹Fe absorption and translocation was observed in plants treated with HEDTA or EDDHA or FOB, but this decrease was avoided in plants treated with HEDTA or EDDHA or FOB in the presence of PS. The enhancement of ⁵⁹Fe absorption and translocation in plants treated with citrate, and the highest ⁵⁹Fe absorption and translocation in plants treated with citrate and PS, indicated that citrate had an additive effect on Fe absorption and translocation in plants. Our results showed that PS effectively played a role in Fe absorption and translocation in plants in the presence of other chelators. Plants treated with any chelators had lower extracellular ⁵⁹Fe in the roots compared with the control.     

Effect of supplied phytosiderophore on 59Fe absorption and translocation in Fe-deficient barley grown hydroponically in low phosphorus media

Abstract

Hydroponically grown barley plants (Hordeum vulgare L. cv. Minorimugi) under iron-deficient (–Fe) and high phosphorus (P) conditions (500 µmol L^?1) showed Fe chlorosis and lower growth compared with plants grown in –Fe and low P conditions (50, 5 and 0.5 µmol L^?1). To understand the physiological role of P in regulating the growth of plants in –Fe medium, we carried out an Fe feeding experiment using four P levels (500, 50, 5 and 0.5 µmol L^?1) and phytosiderophores (PS), mugineic acid. Our results suggest that plants grown in a high P medium had higher absorption activity of ⁵⁹Fe compared with plants grown in low P media, irrespective of the presence or absence of added PS. Translocation of ⁵⁹Fe from roots to shoots was not affected by the P level. The relative translocation rate of ⁵⁹Fe increased with decreasing levels of P in the medium. In general, the addition of PS enhanced the absorption of ⁵⁹Fe and its translocation. Taken together these results suggest that the lower relative translocation rate of Fe in high P plants may be induced by the physiological inactivation of Fe in the roots, and the higher absorption activity of Fe in high P conditions possibly results from the response of barley plants to Fe deficiency.     

Effects of excess manganese and metal chelators on micronutrient concentrations in the xylem sap of iron-deficient barley plants

Barley (Hordeum vulgare L.) plants were grown hydroponically in a greenhouse for 14 d under Fe-deficient conditions before treatment for 3 h with excess Mn (25 µM) and equimolar amounts of plant-borne (phytosiderophores, PS) or synthetic (ethylene diamine tetraacetic acid, EDTA) metal chelators. The xylem sap was collected for 3 h and analyzed for PS, Fe, Mn, Zn, Cu, and citrate concentrations. Excess Mn in the feeding medium decreased the concentrations of PS, Fe, Zn, and Cu in the xylem sap. Addition of 25 µM Mn and an equimolar amount of PS to the feeding medium increased the concentrations of PS, Fe, and Cu in the xylem sap, while EDTA decreased the concentrations of PS and the above nutrients. Excess Mn in the feeding medium increased the Mn concentration in the xylem sap and this increase was more pronounced with the addition of PS to the feeding medium, while EDTA had a depressing effect. These findings suggested that the roots of Fe-deficient barley plants can enhance the absorption and/or translocation of both Mn²⁺ and a PS-Mn complex. Addition of excess Mn to the feeding medium, irrespective of chelators, did not affect the xylem citrate concentration, indicating that citrate may not contribute to the translocation of metal micronutrients. In the xylem sap of Fe-deficient barley plants, the concentrations of metal micronutrients were positively correlated with the concentrations of PSG     

EFFECT OF CADMIUM ON PHYSIOLOGICAL RESPONSES OF WHEAT AND CORN TO IRON DEFICIENCY

This work evaluated the effect of cadmium (Cd) on the physiological responses of corn (Zea Mays L.) and wheat (Triticum aestivum L.) to iron (Fe) deficiency. For this purpose, seedlings of corn and wheat were cultivated under controlled conditions, plants were grown in different strength Hoagland's solutions for one month. In the fifth week, some seedlings were still in full strength Hoagland's solution (+Fe) and others were in full strength Hoagland's solutions without iron (?Fe). The plants were exposed to different cadmium (Cd) concentrations for four days. The plant chlorophyll content of young leaves, Fe and Cd content in shoots and roots, biomass production, and phytosiderophores (PS) release by roots were assessed. Results showed that Cd decreased the chlorophyll content of young leaves, accompanied by a significant shoot and root biomass reduction for Fe-deficient and Fe-sufficient wheat and corn across all Cd treatments. However, chlorophyll content and shoot and root biomass of Fe-deficient wheat and corn were lower than Fe-sufficient plants at different Cd concentrations. Iron-deficiency induced Cd accumulation compared to Fe-sufficient in wheat and corn; however, a depressive effect of Cd on iron acquisition in shoots and roots of Fe-deficient and Fe-sufficient wheat and corn across all Cd treatments was observed. Cadmium also inhibited PS release in Fe-deficient and Fe-sufficient wheat and corn. Iron-deficient PS release was higher than Fe-sufficient corn and wheat across all Cd treatments. These results suggested that Cd might reduce capacity of plants to acquire iron from solution by inhibiting PS release.     

Physiological basis of iron chlorosis tolerance in rice (Oryza sativa) in relation to the root exudation capacity

Micronutrient deficiency in cultivable soil, particularly that of iron (Fe) and zinc (Zn), is a major productivity constraint in the world. Low Fe availability due to the low solubility of the oxidized ferric forms is a challenge. An experiment was, thus, executed to assess the performance of eight genetically diverse rice genotypes on Fe-sufficient (100 µM) and Fe-deficient (1 µM) nutrient solution, and their ability to recover from Fe deficiency was measured. Fe efficiency under Fe deficiency in terms of biomass production showed a significant positive correlation with the root release of phytosiderophore (PS) (R² = 0.62*). This study shows that the Fe deficiency tolerance of Pusa 33 was related to both a high release of PS by the root and an efficient translocation of Fe from the root to the shoot as the Fe–PS complex, which could be useful for improving the Fe nutrition of rice particularly under aerobic conditions.     

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.     

Distribution pattern of root‐supplied 59iron in iron‐sufficient and iron‐deficient bean plants

In order to study the iron (Fe) distribution pattern in bean plants with different Fe nutritional status, french bean (Phaseolus vulgaris L.) seedlings were precultured in a complete nutrient solution with 8x10^‐5 M FeEDTA for five days. Thereafter, plants were further supplied with 8x10^‐5 M FeEDTA (Fe‐sufficient) or with only 2x10^‐6 M FeEDTA (Fe‐deficient) for another eight days. At this stage, the Fe‐deficient plants had much lower chlorophyll contents and lower dry weight of the leaves but higher reducing capacity of the roots compared with the Fe‐sufficient plants. For studies on short‐term distribution of Fe, the Fe‐sufficient plants were supplied 8x10^‐5 M ⁵⁹FeEDTA (specific activity 9.9 GBq/mol) and the Fe‐deficient plants 1x10⁶ M ⁵⁹FeEDTA (specific activity 98.8 GBq/mol). The plants were harvested after 4 and 24 hours. Despite a much lower supply of ⁵⁹FeEDTA/(factor 80), the Fe‐deficient plants took up significantly more ⁵⁹Fe but translocated less to the shoots (14.6% after 24 h) compared with the Fe‐sufficient plants (29.4% after 24 h). However, regardless of the Fe nutritional status of the plants, the majority of ⁵⁹Fe was translocated in the primary leaves. Our results demonstrate a similar distribution patterns of root‐derived ⁵⁹Fe in the shoots of Fe‐sufficient and Fe‐deficient plants, and thus, no preferential direct translocation of Fe to the shoot apex in the Fe‐deficient plants.     

Detection of the regions of phytosiderophore release from barley roots

Abstract

We devised a method to detect regions of phytosiderophore release from barley (Hordeum vulgare cv. Minorimugi) roots. Plants were grown in Fe-sufficient (+Fe) or Fe-deficient (?-Fe) water cultures for 14 d in a phytotron. Intact or excised roots were sampled and put between two sheets of filter papers just after the onset of the light period. The filter papers with roots were wrapped with vinyl film to avoid drying out and covered with aluminum foil to shade. The roots between the filter paper were kept in the phytotron for 4 h and phytosiderophores (PS) released from roots were absorbed by the filter paper. Then, the shape of the roots was preserved by photocopy and PS released on the filter paper were visualized by the method employed for detecting PS on thin layer chromatography (TLC). Gelatinous Fe solubilizing activity was employed for detecting PS on the filter paper. Released PS were detected as white spots on the orange-colored background and the regions of PS release from the roots were visible on the filter paper. It was evident that the apical zones of roots were the main regions of PS release. It was apparent that a distinct primary root newly formed on the basal parts had higher activity to release PS than the other roots of ?Fe plants. It was also shown that apical root zones of +Fe plants released PS.     

Phytosiderophore release in relation to multiple micronutrient metal deficiency in wheat

Phytosiderophore (PS) release, which occurs mainly under iron deficiencies in the representative Poaceae, has been speculated to be a general adaptive response to enhance the acquisition of micronutrient metals. However, it is very common to encounter deficiency of micronutrients other than iron (Fe) in soils and interactions with respect of multi-micronutrient deficiency to effect on PS release are not known. Further, the diurnal rhythm for the release of PS may also be affected under multiple micronutrient deficiency. PS release capacity and PS content of roots and the diurnal rhythm of PS release was measured in selected efficient and inefficient wheat genotypes varied on individual and combined deficiency of Fe, zinc (Zn), copper (Cu) and manganese (Mn) in nutrient solution culture. A nutrient sufficient treatment was also taken as experimental control. Lack of Fe in the nutrient medium caused a significantly higher release of PSs followed by Zn, Mn and Cu in the same order. The diurnal rhythm of PS release was similar in the absence of either of the micronutrients or under their combined deficiency. Micronutrient sufficient control did not release any PS. Fe-use-efficient cultivars produced and released a larger amount of PS and differed from the inefficient cultivars in terms of the PS release but not in the PS biosynthesis in the roots. Thus, indicating that the limitation at the level of release of the PS is responsible for low Fe use efficiency of the Fe deficiency susceptible cultivars. Further, the diurnal variation in the PS release was similar for all the investigated wheat cultivars and did not influence the variation in the Fe use efficiency.     

Diurnal rhythm of release of phytosiderophores and uptake rate of zinc in iron-deficient wheat

Abstract

The diurnal rhythm of release of phytosiderophores and uptake rate of zinc (Zn) was studied in iron (Fe) deficient wheat (Triticum aestivum L. cv. Ares) plants grown in nutrient solution under controlled environmental conditions. Different forms of Zn (e.g. ZnSO₄, ZnEDTA) were used to obtain different degrees of loading of the root apoplasmic pool with Zn.

In the Fe-deficient plants the release of phytosiderophores from the roots followed a distinct diurnal rhythm with a steep peak about 4 h after the onset of the light period. These plants also showed a similar pattern in the rates of Zn uptake over the 24 h day-night cycle. During the light period there was a steep transient peak (factor 3.8) in Zn uptake rate in the Fe-deficient plants supplied with ZnSO₄. This transient peak was much less distinct in plants supplied with ZnEDTA (factor 1.8) and absent in plants supplied with ZnEDTA plus free chelator (+ NaEDTA) in excess. The peak in Zn uptake coincided with the maximum rate of phytosiderophore release in the Fe-deficient plants. In the Fe-sufficient plants the release of phytosiderophores was very low and no such peak in Zn uptake rates could be observed.

These results demonstrate that phytosiderophores mobilize Zn not only in the rhizosphere, but also from the root apoplast. Thus, the apoplasmic pool of micronutrient cations has to be taken into account as potential source for both uptake and diurnal variation in uptake rates of Micronutrient cations.     

Release of phytosiderophores from roots of wheat and triticale under nickel‐deficient conditions

Despite numerous studies on phytosiderophores (PS) there is still an open question whether nickel (Ni) deficiency induces release of PS from graminaceous plant roots. Seedlings of two wheat cultivars (Triticum aestivum L. cvs. Rushan and Kavir) and a triticale cultivar (X. triticosecale) were grown in Ni‐free nutrient solution (Ni‐deficient, Ni–) and with 10 µM NiSO₄ (Ni‐sufficient, Ni+, control). Root exudates were collected weekly for 4 weeks and the amount of PS in the root exudates was measured. The response to Ni deficiency on the release of PS differed between species. Roots of Rushan and triticale exuded higher PS in response to Ni‐deficient conditions. Nickel deficiency significantly enhanced shoot Fe and Zn concentrations in wheat, while it decreased shoot Fe and Zn concentrations in triticale. In Kavir, PS exudation was decreased by Ni deficiency at weeks 3 and 4 and the reduced release of PS from roots of Kavir was accompanied by lower concentrations of Fe and Zn in plant roots but higher Fe and Zn concentrations in shoot tissue. The PS release by Kavir was triggered by a Ni‐induced Zn deficiency particularly in the shoots. According to the results, it is suggested that in the studies concerning the phytosiderophore release under Ni deficiency, special attention should be given to different responses among and within cereals and to the plant Zn or Fe nutritional status.     

Alleviation of Manganese Phytotoxicity in Barley with Calcium

In order to clarify the mechanism by which calcium (Ca) alleviates manganese (Mn) phytotoxicity, barley plants were grown under the following conditions: (1) nutrient solution alone (control), (2) nutrient solution + 25 μM Mn (Mn-toxic), and (3) nutrient solution + 25 μ M Mn + 20 mM Ca (Ca-alleviated). Feeding experiments using ⁵⁴Mn and ⁵⁹Fe (iron) with 2.0 or 20 mM Ca to the plant roots were also conducted. The absorption and translocation of ⁵⁴Mn in the control plants were lowered by the high-Ca (20 mM) feeding condition. The translocation of ⁵⁴Mn to shoots of Mn-toxic or Ca-alleviated plants was also lowered by the high-Ca feeding condition, but ⁵⁴Mn absorption by roots of the plants was unaffected. The absorption and translocation of ⁵⁹Fe in the plants was unaffected by the high-Ca feeding condition. Calcium alleviation of Mn phytotoxicity in barley may be induced mainly by the inhibition of Mn translocation to shoots.     

Effect of supplied phytosiderophore on 59Fe absorption and translocation in Fe-deficient barley grown hydroponically in low phosphorus media

Hydroponically grown barley plants ( Hordeum vulgare L. cv. Minorimugi) under iron-deficient (–Fe) and high phosphorus (P) conditions (500 µmol L⁻¹) showed Fe chlorosis and lower growth compared with plants grown in –Fe and low P conditions (50, 5 and 0.5 µmol L⁻¹). To understand the physiological role of P in regulating the growth of plants in –Fe medium, we carried out an Fe feeding experiment using four P levels (500, 50, 5 and 0.5 µmol L⁻¹) and phytosiderophores (PS), mugineic acid. Our results suggest that plants grown in a high P medium had higher absorption activity of ⁵⁹Fe compared with plants grown in low P media, irrespective of the presence or absence of added PS. Translocation of ⁵⁹Fe from roots to shoots was not affected by the P level. The relative translocation rate of ⁵⁹Fe increased with decreasing levels of P in the medium. In general, the addition of PS enhanced the absorption of ⁵⁹Fe and its translocation. Taken together these results suggest that the lower relative translocation rate of Fe in high P plants may be induced by the physiological inactivation of Fe in the roots, and the higher absorption activity of Fe in high P conditions possibly results from the response of barley plants to Fe deficiency.     

MAIZE HYBRIDS DIFFER IN THEIR 24-H PATTERNS OF PHYTOSIDEROPHORE RELEASE

Iron (Fe) chlorosis tolerant gramineous species respond to Fe-deficiency stress by releasing phytosiderophores. The objective of this study was to characterize the diurnal pattern of phytosiderophore release from Fe-chlorosis tolerant and susceptible maize (Zea mays L.) hybrids. Phytosiderophore was collected from individual, Fe-deficient maize plants in 10 mg L^?1 Micropur solution during 4-h collection periods. The diurnal pattern of phytosiderophore release differed among maize hybrids. Similar levels of phytosiderophore were released during light and dark conditions for three hybrids. Greater phytosiderophore release was measured from a fourth hybrid when the collection procedure began 2-h after the initiation of light compared to 2-h before the initiation of dark. When a single plant was used for six consecutive collection periods, phytosiderophore release declined during the final 8- to 12-h of collection. The decline in phytosiderophore release was attributed to dissolution of apoplastic Fe and deactivation of the Fe-stress response mechanism.     

Iron deficiency stress responses amongst citrus rootstocks

Seedlings of citrus rootstocks differing in lime tolerance were grown in nutrient solution with and without Fe. Proton efflux, release of phenolic compounds and Fe reducing substances and root-mediated reduction of Fe^III in FeEDTA and freshly precipitated Fe(OH)₃ in response to Fe deficiency were determined. Sweet orange, Carrizo citrange and trifoliate orange, the three least tolerant rootstocks used in the study, did not decrease nutrient solution pH in response to Fe deficiency. The more lime tolerant rootstocks, rough lemon, Cleopatra mandarin and sour orange, did decrease nutrient solution pH. But in CaSO₄ solution only sour orange increased H⁺ efflux significantly under Fe deficiency. In response to Fe deficiency, the release of phenolic compounds was increased significantly in rough lemon and Cleopatra mandarin seedlings, while the release of reducing substances was increased significantly in rough lemon, sour orange and trifoliate orange. Rough lemon was the only rootstock to respond to Fe deficiency with an increase in root-mediated reduction of chelated Fe^III at pH 6.5. At pH 8.0, both Fe-deficient rough lemon and Cleopatra mandarin roots reduced higher amounts of Fe^III from freshly precipitated Fe(OH)₃ than Fe-sufficient seedlings. Iron reduction by detached roots of Fe-deficient and Fe-sufficient rough lemon did not follow Michaelis-Menten kinetics at high substrate concentrations. Rates of Fe reduction at low substrate concentrations were inconsistent with the existence of an inducible ferric reductase in response to Fe deficiency.     

Phytosiderophore release from manganese‐induced iron deficiency in barley

An experiment was conducted in the phytotron with barley (Hordeum vulgare L. cv. Minorimugi) grown in nutrient solution to compare iron (Fe) deficiency caused by the lack of Fe with manganese (Mn)‐induced Fe deficiency. Dark brown spots on older leaves and stems, and interveinal chlorosis on younger leaves were common symptoms of plants grown in either Mn‐toxic or Fe‐deficient treatments. Dry matter yield was affected similarly by Fe deficiency and Mn toxicity. The Mn toxicity significantly decreased the translocation of Fe from roots to shoots, caused root browning, and inhibited Fe absorption. The rate of Fe translocated from roots to shoots in the 25.0 μM Mn (toxic) treatment was similar to the Fe‐deficient treatment. Manganese toxicity, based on the release of phytosiderophore (PS) from roots, decreased from 25.0>250>2.50 uM Mn. The highest release of PS from roots occurred 7 and 14 days after transplanting (DAT) to Mn‐toxic and Fe‐deficient treatments, respectively; but was always higher in the Fe‐deficient treatment than the Mn‐toxic treatments. The release of PS from roots decreased gradually with plant age and with severity of the Mn toxicity symptoms. The PS content in roots followed the PS release pattern.     

影响菜豆体内铁再利用效率的因素及其机理

本文在人工气候室中，用营养液培养方法，并结合同位素示踪技术研究了铁的供应状况，两种形态氮素（ＮＯ＾－３－Ｎ和ＮＨ＾＋４－Ｎ）及叶处遮光对菜豆体内铁再利用效率的影响，并对其有关机理进行了深入的研究。结果表明，铁的缺乏有利于累积在根和初生叶中的铁身新生组织中转移，铁的再利用效率明显提高。无论有缺铁还供铁条件下，ＮＨ＾＋４－Ｎ的供应使得菜豆新叶中活性铁含量、新叶叶绿素含量及体内铁的再利用效率都明显高于Ｎ     

Water- and pyrophosphate-extractable humic substances fractions as a source of iron for Fe-deficient cucumber plants

The capacity of Fe-deficient cucumber plants to utilise water-extractable and pyrophosphate-extractable humic substances as a source of Fe was investigated. Plants were grown for 13 days in nutrient solution in the presence or absence of Fe and during the last 7 days water-extractable and pyrophosphate-extractable humic substances were added to the solution at a final concentration of 5 μg organic C ml^–1. The water-extractable humic fraction did not significantly modify leaf area and dry matter accumulation, leaf total Fe or chlorophyll content of cucumber plants adequately supplied with Fe. In contrast, pyrophosphate-extractable humic substances caused a slight but significant decrease of all the leaf parameters considered, with the exception of the chlorophyll content. Root Fe content of Fe-sufficient plants was decreased by more than 50% in the presence of each humified fraction. Addition of each humic fraction to Fe-deficient plants led to a partial disappearance of leaf chlorosis symptoms with a significant increase in chlorophyll and leaf Fe content. Fe content of roots was also significantly increased in Fe-deficient plants by the addition of humic substances to the nutrient solution. These results show that Fe-deficient cucumber plants can utilise Fe contained in the two fractions of humified organic matter. However, by calculating the amount of total Fe accumulated per plant in the presence of water-extractable or pyrophosphate-extractable humic substances, it could be seen that Fe contained in the water-extractable humic fraction was almost totally used by Fe-deficient cucumber plants, while that present in the pyrophosphate-extractable fraction could only be partially absorbed. The results strongly support a role of humified organic matter in Fe nutrition of plants and are discussed in terms of a possible interaction between soil humic substances and the biochemical mechanisms involved in the plant response to Fe deficiency. Received: 6 November 1996     

Arsenic–iron interaction: Effect of additional iron on arsenic-induced chlorosis in barley grown in water culture

Abstract

The effect of additional iron (Fe) on arsenic (As) induced chlorosis in barley (Hordeum vulgare L. cv. Minorimugi) was investigated. The treatments were: (1) 0?μmol?L^?1 As?+?10?μmol?L^?1 Fe³⁺ (control), (2) 33.5?μmol?L^?1 As?+?10?μmol?L^?1 Fe³⁺ (As-treated) and (3) 33.5?μmol?L^?1 As?+?50?μmol?L^?1 Fe³⁺ (additional-Fe³⁺) for 14?days. Arsenic and Fe³⁺ were added as sodium-meta arsenite (NaAsO₂) and ethylenediaminetetraacetic acid-Fe³⁺, respectively. Chlorosis in fully developed young leaves was observed in the As-treated plants. The chlorophyll index and the Fe concentration decreased in shoots of the As-treated plants compared with the control plants. Arsenic reduced the concentration of phosphorus, potassium, calcium, magnesium, manganese, zinc and copper. The additional-Fe³⁺ treatment increased the chlorophyll index in plants compared with the As-treated plants. Among the elements, Fe concentration and accumulation specifically increased in the shoots of additional-Fe³⁺ plants compared with As-treated plants, indicating that As-induced chlorosis was Fe-chlorosis. Arsenic and Fe were mostly concentrated in the roots of the As-treated plants. Despite inducing chlorosis in the As-treated plants, phytosiderophores (PS) accumulation in the roots and release from the roots did not increase, rather PS accumulation decreased, indicating that As toxicity hindered PS production in the roots. The PS accumulation in the roots was further reduced in the additional-Fe³⁺ treatment.