Diurnal variations in absorption and translocation of a ferrated phytosiderophore in barley as affected by iron deficiency

The release of phytosiderophore (PS) from roots of Fe-deficient graminaceous plants follows a distinct diurnal rhythm with maximum release rates occurring usually 3 to 4 hours after the onset of light. However, it remains to be determined whether absorption of the PS-Fe³⁺ complex shows a diurnal rhythmicity similar to that of PS release, Barley plants grown with or without 10 µM FeEDTA for 7 days were fed with ferreted PS (10 µM labelled with ⁵⁹Fe) at 4-h intervals to study the diurnal variations in the absorption and transloca tion of ⁵⁹Fe, The absorption of ⁵⁹Fe, irrespective of the Fe nutritional status of the plants, was higher during the day and lower during the night but did not show any peak throughout the day-night cycle. On the other hand, the translocation of ⁵⁹Fe into shoots of Fe-deficient plants was lower than that of Fe-sufficient plants, while the Fe nutritional status of the plants did not affect the absorption of ⁵⁹Fe by roots, The formation of root apoplastic ⁵⁹Fe was lower during the day and higher during the night, regardless of the Fe nutritional status of plants. Our results showed that the absorption of the PS-Fe³⁺ complex by roots did not follow the PS release pattern.     

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.     

Alien cytoplasm effects on phytosiderophore release in two spring wheats (Triticum aestivum L.)

Iron (Fe) deficiency is a difficult nutrient problem particularly in crop plants grown on calcareous soils. Recently, phytosiderophore (PS) release has been linked to the ability of graminaceous species and genotypes to withstand Fe-deficiency chlorosis. So enhancing PS release is a critical step to improve iron efficiency of plants grown on iron stressed soils. The effects of alien cytoplasm on PS release in spring wheat were studied by analyzing PS release from twenty wheat genotypes, including two spring wheat 881 and 352-35, and their 18 alloplasmic lines with the participation of cytoplasms from the Aegilops and Triticum species. Different genotypes were grown in iron sufficient and deficient nutrient solution under controlled environmental conditions. PS release rates were determined at two or three days intervals after onset of iron deficiency symptoms by the measurement of iron mobilizing capacity of root exudates from freshly precipitated Fe^III hydroxide. High amounts of phytosiderophores were released from roots of all wheat genotypes without iron supplied, and the amount progressively increased with the development of iron deficiency chlorosis. The results revealed that (1) the release rate of phytosiderophores from roots of common wheat could be considerably influenced by alien cytoplasms. Some alien cytoplasms exerted positive effects, some ones did negative effects, and the other ones had no significant effects. (2) the same alien cytoplasm could affect similarly or oppositely the phytosiderophores release from different wheat. (3) some alien cytoplasms, such as Chinese Spring, Ae. speltoides Tausch and Ae. cylindrica Host showed promising and potential in improving the rate of phytosiderophore release in common wheat. These cytoplasms which showed the desired effect should be given priority in interspecific and intergeneric hybridization to develop and reconstruct the needed wheat cultivars.     

Alleviation of manganese toxicity and manganese-induced iron deficiency in barley by additional potassium supply in nutrient solution

Barley plants were grown hydroponically at two levels of K (3.0 and 30 mm) and Fe (1.0 and 10 μm) in the presence of excess Mn (25 μm) for 14 d in a phytotron. Plants grown under adequate K level (3.0 mm) were characterized by brown spots on old leaves, desiccation of old leaves, interveinal chlorosis on young leaves, browning of roots, and release of phytosiderophores (PS) from roots. These symptoms were more pronounced in the plants grown under suboptimal Fe level (1.0 p,M) than in the plants grown under adequate Fe level (10 μm). Plants grown in 10 μm Fe with additional K (30 mm) produced a larger amount of dry matter and released less PS than the plants grown under adequate K level (3.0 mm), and did not show leaf injury symptoms and root browning. On the other hand, the additional K supply in the presence of 1.0 μM Fe decreased the severity of brown spots, prevented leaf desiccation, and increased the leaf chlorophyll content, which was not sufficient for the regreening of chlorotic leaves. These results suggested that the additional K alleviated the symptoms of Mn toxicity depending on the Fe concentration in the nutrient solution. The concentration (per g dry matter) and accumulation (per plant) of Mn in shoots and roots of plants grown in 10 μm Fe and 30 mm K were much lower than those of the plants grown in 10 μm Fe and 3.0 mm K, indicating that additional K repressed the absorption of Mn. The concentration and accumulation of Fe in the shoots and roots of the plants grown in 10 μm Fe and 30 mm K were higher than those of the plants grown in 10 μm Fe and 3.0 mm K, indicating that the additional K increased the absorption of Fe under excess Mn level in the nutrient solution. The release of PS, chlorophyll content, and shoot Fe concentration were closely correlated.     

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.     

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.     

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.     

Genotypic variation in Fe concentration of barley grain

Abstract

Iron deficiency in humans is the most prevalent micronutrient deficiency worldwide. To screen for high Fe cultivar, genotypic variation in Fe concentration of barley grain was investigated in two collections of barley: 274 standard varieties selected at the Barley Germplasm Center of the Research Institute for Bioresources, Okayama University (SV) and 135 varieties from the Barley Core Collection of Americas (BCCAM). The Fe concentration of barley grain showed large variation, ranging from 24.6 to 63.3 mg kg^?1 in SV, from 21.0 to 83.0 mg kg^?1 in BCCAM barleys. The Fe concentration was not affected by the key characters of barley varieties, kernel row types, and hulled or hull-less. The Fe concentration was also not related to the place of origin of the barley variety. About 90% of total Fe was localized in the grain without hull. These results provide fundamental data for breeding Fe-rich cultivars and for studying the mechanisms involved in genotypic variation in Fe concentration.     

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     

Comparison of the functions of the barley nicotianamine synthase gene HvNAS1 and rice nicotianamine synthase gene OsNAS1 promoters in response to iron deficiency in transgenic tobacco

Barley ( Hordeum vulgare L.) nicotianamine synthase gene ( HvNAS1 ) expression in barley is strongly induced by Fe deficiency in the roots and rice ( Oryza sativa L.) nicotianamine synthase gene ( OsNAS1 ) expression in rice is induced by Fe deficiency both in the roots and in the shoots. In dicots, NAS genes are not strongly induced by Fe deficiency, and they function to maintain Fe homeostasis. Rice OsNAS1promoter::GUS or barley HvNAS1promoter::GUS was introduced into tobacco ( Nicotiana tabacum L.) and tissue specificities and systemic regulation of their expression were compared. A split-root experiment revealed that the HvNAS1 promoter exhibited functions similar to those of Fe-acquisition-related genes in tobacco roots, suggesting that this promoter responded to certain Fe-deficiency systemic signals and to the Fe concentration in the rhizosphere. The HvNAS1 promoter might harbor a type of universal system of gene expression for Fe acquisition. However, the OsNAS1 promoter did not respond to local application of Fe to the roots and induced GUS activities in mature leaves in response to Fe deficiency. This promoter might possess numerous types of cis -acting sequences that are involved in Fe metabolism.     

Comparison of iron chelates and complexes supplied as foliar sprays and in nutrient solution to correct iron chlorosis of soybean

The application of synthetic chelates is the most efficient remedy for correcting iron (Fe) chlorosis. However, chelates are usually expensive and nondegradable products. Recently, new degradable chelates have been proposed for their use as Fe fertilizers. Also, Fe complexes cheaper than synthetic chelates and derived from natural products are also used to correct Fe deficiencies. Fifteen products, including five different synthetic chelates (Fe‐EDDS, Fe‐IDHA, and three Fe‐EDTA formulations) and ten natural complexes (humates, lignosulfonates, amino acids, glycoproteins, polyamines, citrate, and gluconate), have been compared when applied at low concentration to soybean (Glycine max L.) chlorotic plants grown in hydroponics under controlled conditions. In the first experiment, Fe compounds were applied to the nutrient solution, while in the second trial, Fe was foliar‐supplied. Dry matter, Fe concentration in shoots and roots, and SPAD values were used to evaluate the effectiveness of the Fe in the different products. In the nutrient‐solution experiment, synthetic chelates provided better plant growth, Fe concentration, and SPAD values than complexes. Among the Fe complexes, transferrin generally provided good plant responses, similar to those obtained with synthetic chelates. After foliar application, the highest regreening was observed for plants treated with synthetic chelates and amino acid complexes, but the translocation to roots only occurred for Fe lignosulfonate. Fe‐EDDS and Fe‐EDTA performed in a similar way when applied in nutrient solution or as foliar sprays.     

Efficacy of an artificial microbial siderophore-Fe(III) with high redox potential on correcting Fe chlorosis in rice

ABSTRACT

Microbial siderophore-chelated Fe(III) is suggested to be an important source of Fe for plants, although it is hardly reduced by plant roots. Here, we investigated the efficacy of the easily reducible artificial microbial siderophore tris[2-{(N-acetyl-N-hydroxy)glycylamino}ethyl]amine (TAGE)-Fe(III) as an alternative Fe source to correct Fe deficiency in rice plants, and compared it to that of the natural siderophore deferoxamine B (DFOB)-Fe(III). We also evaluated the absorption of Fe from TAGE-Fe(III) by the Strategy I-like system of gramineous plants using nicotianamine aminotransferase 1 (naat1) mutant rice, which does not synthesize phytosiderophores. Fe(III)-siderophores were synthesized in vitro. Nipponbare rice and its naat1 mutant were reared in soil and gel cultures to determine Fe availability. Hydroponically grown naat1 mutant seedlings were used for reducibility assays to determine the ability of rice roots to reduce Fe(III) chelated by TAGE or DFOB. The expression of a Fe-deficiency inducible gene was also determined, as well as chlorophyll and Fe concentrations. Reduci bility assays on naat1 mutant seedlings revealed that the reduction level of TAGE-Fe(III) was approximately three times higher than that of DFOB-Fe(III). Application of TAGE-Fe(III) to both culture medium and alkaline soil improved Fe chlorosis, growth, and Fe concentration in both naat1 and wild type plants, whereas application of DFOB-Fe(III) only did so in wild type plants. Easily reducible Fe(III)-chelates such as TAGE-Fe(III) can be a better source of Fe for rice plants than most natural microbial siderophores-Fe(III). Our study also demonstrated that rice plants have the ability to utilize microbial siderophores-Fe(III) as the Fe source through the Strategy I-like Fe acquisition system.     

Fine roots for uptake and translocation of zinc into young leaves as important traits for zinc efficiency of maize genotypes

Suitable germplasm and breeding for zinc (Zn)-efficient genotypes has been prioritized to combat Zn deficiency problem. Six maize genotypes were grown in a nutrient solution with or without Zn to identify genotype tolerance to low Zn and its mechanism. Zhongnong 99 was found to be about 2-fold more Zn-efficient than L69 × 178. Young shoot Zn concentration and content was significantly correlated with Zn efficiency (ZE). Furthermore, Zn-efficient genotype as Zhongnong 99 distributed more Zn into young parts of shoot and also developed longer and thinner roots (≤ 0.2 mm) under -Zn conditions than inefficient genotypes. The close relationship (0.82***, 0.88***, 0.90***) between Zn content of young shoots and fine roots indicated that fine roots (≤ 0.2 mm) contributed more efficient Zn uptake into old parts which distributed to young parts. In conclusion, the analysis of young shoots and fine roots (≤ 0.2 mm) represent a suitable screening technique for ZE evaluation of under -Zn conditions.     

Zinc Deficiency in Selected Cultivars of Wheat and Barley as Tested in Solution Culture

Abstract

The relative zinc (Zn) efficiencies of 33 wheat and 3 barley cultivars were determined by growing them in chelate‐buffered culture solutions. Zn efficiency, determined by growth in a Zn‐deficient solution relative to that in a medium containing an adequate concentration of Zn, was found to vary between 10% and 63% among the cultivars tested. Out of the 36 cultivars tested, 12 proved to be Zn efficient, 10 were Zn inefficient, and the remaining 14 varieties were classed as intermediate. The most Zn‐efficient cultivars included Bakhtawar, Gatcher S61, Wilgoyne, and Madrigal, and the most Zn inefficient included Durati, Songlen, Excalibur, and Chakwal‐86. Zn‐efficient cultivars accumulated greater amounts of Zn in their shoots than inefficient cultivars, but the correlation between shoot Zn and shoot dry matter production was poor. All the cultivars accumulated higher concentrations of iron (Fe), copper (Cu), manganese (Mn), and phosphorus (P) at deficient levels of Zn, compared with adequate Zn concentrations. The Zn‐inefficient cultivars accumulated higher concentrations of these other elements compared to efficient cultivars.     

Incorporation of 14CO2 and 15NH4 into amino acids of the two subunits of fraction 1 protein in spinach leaves

In the investigation of nitrogen metabolism in plants, it is important to deal with proteins, which are the end-products of nitrogen metabolism.     

Importance of iron use‐efficiency of nodules in common bean (Phaseolus vulgaris L.) for iron deficiency chlorosis resistance

Several studies suggest that the Fabaceae‐Rhizobium symbiosis is particularly sensitive to iron (Fe) deficiency with respect to NO₃^–‐dependent plants. The aim of this study, which is part of a screening program for common bean tolerance to Fe deficiency, was to study genotypical differences in Fe requirement and Fe use‐efficiency of common bean cultivars depending on symbiotic nitrogen fixation (SNF). Results show that ARA14 produces more whole plant dry matter and particularly more nodule biomass than Coco blanc. ARA14 is characterized by a high capacity of nitrogen fixation and a better Fe use‐efficiency for the growth and the function of the nodules.     

盆栽研究钢渣用作铁肥对玉米生长和土壤改良的作用

The feasibility of steel slag used as an iron fertilizer was studied in a pot experiment with corn. Slag alone or acidified slag was added to two Fe-deficient calcareous soils at different rates. Results showed that moderate rates （10 and 20 g kg^-1） of slag or acidified slag substantially increased corn dry matter yield and Fe uptake. Application of steel slag increased the residual concentration of ammonium bicarbonate-diethylenetriamine pentaacetic acid （AB-DTPA） extractable Fe in the soils. The increase of extractable Fe was usually proportional to the application rate, and enhanced by the acidification of slag. Steel slag appeared to be a promising and inexpensive source of Fe to alleviate crop Fe chlorosis in Fe-deficient calcareous soils.     

Incorporation of S into soil organic matter in the field as determined by the natural abundance of stable S isotopes

In agricultural systems with low S inputs, soil organic matter is a major source of S and the transformations between organic and inorganic S pools are important for the supply of S to plants. This study was conducted to determine the effect of S fertilizer on the size and activity of organic S pools. For 5 years S fertilizer with a known composition of stable S isotopes was applied to a rotation on a loamy soil and a coarse sandy soil at rates higher than the plant demand. Total organic S in soil organic matter was not affected by sulphur application, but a small increase occurred in the sulphate ester fractions (P<0.05). Inorganic sulphate concentrations in the soil reflected the S application in the year of sampling, whereas S applied in earlier years was not recognized. Organic matter below the plough layer in both soils was enriched with S, possibly as a result or organic matter leaching or an increased clay content in the subsoils. At 0–20 cm, the C:S ratio in organic matter was ca. 100 for both soils, decreasing to 73 and 46 at 60–80 cm for the coarse sandy soil and the loamy soils, respectively. In both soils, isotope data showed that ca. 30% of organic-bonded S at 0–20 cm originated from fertilizer S applied during the last 5 years, irrespective of the S application rate. At 20–40 cm the rate of incorporations was lower and at 40–60 cm no incorporation of fertilizer S into organic matter was recognized. The fertilizer application did not induce net changes in the total organic S fraction, but isotope data indicated that a considerable part of the organic S pool was involved in S cycling in the field.     

Depressed growth of non‐chlorotic vine grown in calcareous soil is an iron deficiency symptom prior to leaf chlorosis

The development of iron deficiency symptoms (growth depression and yellowing of the youngest leaves) and the distribution of iron between roots and leaves were investigated in different vine cultivars (Silvaner, Riparia 1G and SO4) grown in calcareous soils. As a control treatment all cultivars were also grown in an acidic soil. Only the cultivars Silvaner and Riparia 1G showed yellowing of the youngest leaves under calcareous soil conditions at the end of the cultivation period. All cultivars including SO4 showed severe shoot growth depression, by 50 % and higher, before yellowing started or without leaf yellowing in the cultivar SO4. Depression of shoot growth occurred independently from that of root growth. In a further treatment the effect of Fe‐EDDHA spraying onto the shoot growth of the cultivar Silvaner after cultivation in calcareous soil was investigated. Prior to Fe application plants were non‐chlorotic, but showed pronounced shoot growth depression. Spraying led to a significant increase in shoot length, though leaf growth was not increased. Accordingly, depression of shoot growth of non‐chlorotic plants under calcareous soil conditions and with ample supply of nutrients and water has been evidenced to be at least partly an iron deficiency symptom. We suggest that plant growth only partially recovered because of dramatic apoplastic leaf Fe inactivation and/ or a high apoplastic pH which may directly impair growth. Since growth was impaired before the youngest leaves showed chlorosis we assume that meristematic growth is more sensitively affected by Fe deficiency than is chlorophyll synthesis and chloroplast development. In spite of high Fe concentrations in roots and leaves of the vines grown in calcareous soils plants suffered from Fe deficiency. The finding of high Fe concentrations also in young, but growth retarded green leaves is a further indication that iron deficiency chlorosis in calcareous soils is caused by primary leaf Fe inactivation. However, in future, only a rigorous study of the dynamic changes of iron and chlorophyll concentration, leaf growth and apoplastic pH at the cellular level during leaf development and yellowing will provide causal insights between leaf iron inactivation, growth depression, and leaf chlorosis.<?show $6#>