Release of zinc mobilizing root exudates in different plant species as affected by zinc nutritional status

The effect of zinc nutritional status of the plant on the release of zinc mobilizing root exudates was studied in various dicotyledonous (apple, bean, cotton, sunflower, tomato) and graminaceous (barley, wheat) plant species grown in nutrient solutions. In all species, zinc deficiency increased root exudation of amino acids, sugars and phenolics. However, the root exudates of zinc deficient dicotyledonous species did not enhance zinc mobilization from a synthetic resin (Zn chelite), or a calcareous soil, although mobilization of iron from Fe^III hydroxide was increased. By contrast in the graminaceous species, root exudates from zinc deficient plants greatly increased mobilization of both zinc and iron from the various sources. These differences in capability of mobilization of zinc and iron between the plant species are the result of an enhanced release of phytosiderophores with zinc deficiency in the graminaceous species.     

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.     

Iron deficiency‐induced zinc uptake by bread wheat

The present study aimed to test the contribution of the iron (Fe) deficiency‐induced uptake system to zinc (Zn) and copper (Cu) uptake by using bread wheat (Triticum aestivum cv. Bezostaja). For this purpose, two different uptake experiments, long‐term and short‐term, were set up in a nutrient solution culture under controlled growth chamber conditions. For the long‐term experiment, wheat cv. plants were grown with different concentrations of Fe or Zn. Results show that there was an uptake system induced under Fe‐limiting conditions which also contributed to Zn and Cu uptake. However, the Zn deficiency‐induced uptake mechanism affected neither Fe nor Cu uptake by wheat. Short‐term uptake experiments indicate that Fe deficiency‐induced Zn²⁺ uptake was more enhanced than the absorption of Zn‐phytosiderophore (PS) complexes. In addition, the Fe‐deficient plants absorbed more Zn in comparison to those plants supplied with sufficient Fe. Similar tendencies in Zn uptake under Fe deficiency in both short‐ and long‐term experiments suggest that there may be a specific Fe uptake system induced under Fe‐limiting conditions for non‐chelated metals in bread wheat. Moreover, this system also contributes to the transport of inorganic forms of some other metals, such as Zn and Cu. Although evidence is still needed involving the use of molecular biological techniques, it is hypothesized that IRT‐like proteins are responsible for this uptake system. Moreover, the release of Fe deficiency‐induced phytosiderophores and uptake of Fe(III)‐phytosiderophore complexes may not be the only mechanisms involved in the adaptation of wheat to Fe‐limiting conditions.     

Phytosiderophore release by Sorghum,wheat, and corn under zinc deficiency 1

Zinc (Zn) deficiency is more common in corn (Zea mays L.) than in sorghum [Sorghum bicolor (L.) Moench] or wheat (Triticum sp.). The ability of wheat to withstand low soil Zn conditions is related to increased release of phytosiderophore from its roots. The reasons for sorghum's ability and corn's inability to utilize low levels of soil Zn have not been explored adequately. The objectives of this research were to 1) ascertain if Zn deficiency could be induced in sorghum, wheat, and corn grown in a chelator‐buffered nutrient solution and 2) determine relative releases of phytosiderophore from roots of sorghum, wheat, and/or corn under Zn‐deficiency conditions. Sorghum, wheat, and corn were grown hydroponically in the greenhouse with a chelator‐buffered nutrient solution designed to induce Zn deficiency, while supplying adequate amounts of other nutrients. Root exudates were collected over time to measure phytosiderophore release. Shoot Zn concentrations and shoot and root dry matter yields were determined also. The technique was effective for inducing Zn deficiency in sorghum, wheat, and corn, as evidenced by reduced shoot and root dry matter yields, shortened internodes, reduced shoot Zn concentrations, and plant Zn concentrations below the suggested critical values for these species. Sorghum and wheat plants increased the release of phytosiderophore in response to Zn deficiency, but com did not. The total amount of phytosiderophore released by the roots was in the order wheat>sorghum>corn. The absence of a “phytosiderophore”; response to Zn deficiency of corn, coupled with the evidence that this species requires, or at least accumulates, more Zn than wheat or sorghum, provides an explanation as to why Zn deficiencies are more prevalent for corn than wheat or sorghum under field conditions.     

Phytosiderophore release does not relate well with zinc efficiency in different bread wheat genotypes

Using six bread wheat genotypes (Triticum aesttvum L. cvs. Dagdas‐94, Gerek‐79, BDME‐10, SBVD 1–21, SBVD 2–22 and Partizanka Niska) and one durum wheat genotype (Triticum durum L. cv. Kunduru‐1149) experiments were carried out to study the relationship between the rate of phytosiderophore release and susceptibility of genotypes to zinc (Zn) deficiency during 15 days of growth in nutrient solution with (1 μM Zn) and without Zn supply. Among the genotypes, Dagdas‐94 and Gerek‐79 are Zn efficient, while the others are highly susceptible to Zn deficiency, when grown on severely Zn deficient calcareous soils in Turkey. Similar to the field observations, visual Zn deficiency symptoms, such as whitish‐brown lesions on leaf blades occurred first and severely in durum wheat Kunduru‐1149 and bread wheats Partizanka Niska, BDME‐10, SBVD 1–21 and SBVD 2–22. Visual Zn deficiency symptoms were less severe in the bread wheats Gerek‐79 and particularly Dagdas‐94. These genotypic differences in susceptibility to Zn deficiency were not related to the concentrations of Zn in shoots or roots. All bread wheat genotypes contained similar Zn concentration in the dry matter. In all genotypes supplied adequately with Zn, the rate of phytosiderophore release was very low and did not exceed 0.5 μmol/48 plants/ 3 h. However, under Zn deficiency the release of phytosiderophores increased in all bread wheat genotypes, but not in the durum wheat genotype. The corresponding rates of phytosiderophore release in Zn deficient durum wheat genotype were 1.2 umol and in Zn deficient bread wheat genotypes ranged between 8.6 μmol for Partizanka Niska to 17.4 umol for SBVD 2–22. In Dagdas‐94, the most Zn efficient genotype, the highest rate of phytosiderophore release was 14.8 umol. The results indicate that the release rate of phytosiderophores does not relate well with the susceptibility of bread wheat genotypes to Zn deficiency. Root uptake and root‐to‐shoot transport of Zn and particularly internal utilization of Zn may be more important mechanisms involved in expression of Zn efficiency in bread wheat genotypes than release of phytosiderophores.     

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.     

Role of root-induced changes in the rhizosphere for iron acquisition in higher plants

Plants can mobilize iron (Fe) in the rhizosphere by non-specific and specific (adaptive) mechanisms. Non-specific mechanisms are, for example, rhizosphere acidification related to high cation-anion uptake ratios, or citric acid excretion. The specific mechanisms are root responses to Fe deficiency and can be classified into two different strategies. The Strategy I is typical for dicots and monocots except for grasses (graminaceous species) and is characterized by increased plasma membrane-bound reductase activity, enhanced net excretion of protons and enhanced release of reducing compounds, mainly phenolics. The reductase activity is stimulated by low pH, and with supply of Fe^III chelates, ferric reduction at the plasma membrane takes place prior to uptake. In contrast, in graminaceous species (Strategy II) these root responses are absent, but enhancement of release of Fe^III chelating compounds - phytosiderophores - takes place. These phytosiderophores are very efficient in mobilizing Fe^III from artificially prepared sparingly soluble inorganic compounds (e.g. Fe^III hydroxide) and from calcareous soils. The ferrated phytosiderophores are taken up by grasses at rates 10² to 10³ times higher than Fe supplied either as synthetic chelate or microbial siderophores (e.g. ferrioxamine B), indicating a specific membrane transport system for ferrated phytosiderophores in roots of grasses. In calcareous soils phytosiderophores not only mobilize Fe, but also Zn, Mn, and Cu by chelation. However, only the Fe^III phytosiderophores are taken up preferentially by Fe deficient grasses. The ecological advantages and disadvantages of Strategy I and Strategy II for Fe acquisition from calcareous soils are discussed.     

Zinc‐efficient wild grasses enhance release of phytosiderophores under zinc deficiency

The effect of the zinc (Zn) nutritional status on the rate of phyto‐siderophore release was studied in three wild grass species (Hordeum murinum, Agropyron orientale, and Secale cereale) grown in nutrient solution under co‐trolled environmental conditions. These wild grasses are highly “Zn‐efficient”; and grow well on severely Zn‐deficient calcareous soils in Turkey (DTPA‐extractable Zn was 0.12 mg/kg soil and CaCO₃ was 37%). In all wild grasses studied, Zn deficiency reduced shoot growth but had no effect on root growth. Low amounts of phytosiderophores were released from roots of all wild grasses adequately supplied with Zn. In plants grown without Zn, release of phytosiderophores progressively increased with the onset of visual Zn deficiency symptoms, such as inhibition of shoot elongation and appearance of chlorotic and necrotic patches on leaves. Compared to Zn‐sufficient plants, phytosiderophore release increased 18–20‐fold in deficient plants. HPLC analysis of root exudates showed that the dominating phytosiderophore in Zn‐deficient Agropyron and Hordeum was 3‐epi‐hydro‐xymugineic acid (epi‐HMA) and was 3‐hydroxy‐mugineic acid (HMA) in Secale. Besides HMA, epi‐HMA and mugineic acid (MA) were also detected in exudates of Zn‐deficient Secale. The results indicate the importance of phytosiderophores in adaptation of wild grasses to Zn‐deficient calcareous soils. Phytosiderophores might enhance mobilization of Zn from sparingly soluble Zn pools and from adsorption sites, both in the rhizosphere and within the plants.     

Iron inefficient and efficient oat cultivars. II. Characterization of phytosiderophore released in response to Fe deficiency stress

Abstract

Iron‐inefficient TAM 0–312 and Fe‐efficient Coker 227 oats (Strategy II plants) differ in their release of phytosiderophore in response to iron‐deficiency stress—the Fe‐efficient Coker 227 releases a phytosiderophore whereas the Fe‐inefficient TAM 0–312 does not. The phytosiderophore released by Coker 227 oats in response to Fe‐deficiency stress does not appear to transport Fe into the plant as Fe phytosiderophore. When the Fe‐inefficient TAM 0–312 and Fe‐efficient Coker 227 oats were subjected to Fe supplied as Fe²⁺(BPDS)₃, Fe³⁺HEDTA, as Fe³⁺EDDHA, Coker 227 utilized the Fe more efficiently than TAM 0–312 in every case. Both cultivars reduced Fe³⁺ as FeCl₃ to form Fe²⁺(BPOS)₃ and responded better to this form of Fe than Fe supplied as the ferric chelate. Reduction of Fe³⁺ at the root appears to be a factor that facilitates iron uptake by Coker 227 oats and the release of a phytosiderophore appears to make more Fe available at the root that can be reduced and transported to plant tops.     

根分泌物与禾本科植物对缺铁胁迫的适应机理

本文系统地总结了自然基金重点项目根分泌物在根际微生态系统中的营养机理的研究进展和部分主要成果。研究表明,养分专一性根分泌物是植物营养遗传特性控制基因的标记物,它受某一养分缺乏的诱导,是在植物体内合成并可通过主动分泌作用进入根际的代谢产物。它的合成和分泌只受该养分胁迫的专一诱导和控制,只要改善这一营养状况就能抑制或阻止其合成和分泌。当植物缺乏这一养分时,植物体可通过自身的调节能力,合成专一性物质并自根分泌到根际,促进该养分的活化并提高植物对其吸收利用效率,从而达到克服或缓解该营养胁迫的目的。用单基因突变材料进行的研究表明,植物铁载体的生物合成和吸收利用是受单基因控制的过程。这一发现不仅使人类有可能运用生物学研究技术来解决营养缺乏问题,而且也为有效地利用自然资源、降低生产成本、减少环境污染提供了可能性。麦根酸类(mugineic.acids)植物铁载体(phytosiderophores)在缺铁禾本科植物体内的生物合成,从植物根内向根际的分泌、在根际环境中对铁的活化及植物对F63+植物铁载体螯合体的吸收四个过程组成了禾本科植物对缺铁胁迫的适应机理。植物铁载体只在早晨日出后2～6h内大量分泌的节律性增加了它们在根际土壤中的相对浓度,减少了它们与土壤颗粒的接触和被吸附;分泌部位集中在微生物尚未侵染的根尖避开了微生物的破坏和分解,同时也增加了它们在土壤微区中的相对浓度。分泌作用和螯合作用不受介质pH值和Ca2+离子浓度影响的特性使该机理在经常出现缺铁现象的石灰性土壤上具有特殊意义。研究结果还表明,缺铁可以诱导激活根细胞原生质膜上可能存在的Fe3+植物铁载体复合体的专一性吸收和运载蛋白,高pH值和CaCO3对这一蛋白的载体功能只有很小的抑制作用,即使在pH值和CaCO3含量都较高的石灰性土壤上,也能有效地发挥作用。这一结论揭示了小麦等禾本科植物适应铁胁迫的实质及其生态学意义。     

Factors in iron‐stress response mechanism enhanced by Zn‐deficiency stress in Sanilac,but not Saginaw navy bean

Zinc‐inefficient Sanilac and Zn‐efficient Saginaw navy bean (Phaseolus vulgaris L.) differ in their susceptibility to Zn‐deficiency stress. Sanilac accumulates Fe under Zn‐deficiency stress and Saginaw does not. These two navy bean cultivars were grown at 0, 0.006 and 0.12 mg/L Zn in modified Hoagland nutrient solution. Various Fe‐stress response mechanisms were quantified periodically over a 12‐day experimental period to determine if known factors in the Fe‐stress response mechanism were enhanced by Zn‐deficiency stress. Visual Zn‐deficiency symptoms were more severe in Sanilac than Saginaw navy bean under equivalent Zn treatments. Sanilac contained lower leaf Zn than Saginaw when Zn was present in solution (0.006 and 0.12 mg/L Zn), but the two cultivars were similar in leaf Zn in the absence of Zn (0 mg/L Zn). Sanilac accumulated more leaf Fe than Saginaw when under Zn stress (0 and 0.006 mg/L Zn). The higher levels of leaf Fe in Sanilac than Saginaw were closely associated with enhanced release of reductants and increased reduction of Fe³⁺ to Fe²⁺ by roots of Sanilac. Saginaw navy bean roots reduced Fe³⁺ to Fe²⁺ similarly to Sanilac with adequate Zn present in solution (0.12 mg/L), but experienced minuscule levels of Fe³⁺ reduction under Zn deficiency. Zinc deficiency stimulated the initiation of the Fe‐stress response mechanism in Sanilac, but not Saginaw, which may have enhanced the development of Zn‐deficiency symptoms in Sanilac due to the increased uptake of Fe by this cultivar. The common Fe‐deficiency stress response associated primarily with grasses (release of phytosiderophore) was not found in either navy bean cultivar.     

Root exudation of sugars,amino acids,and organic acids by maize as affected by nitrogen,phosphorus, potassium,and iron deficiency

Root exudates play a major role in the mobilization of sparingly soluble nutrients in the rhizosphere. Since the amount and composition of major metabolites in root exudates from one plant species have not yet been systematically compared under different nutrient deficiencies, relations between exudation patterns and the type of nutrient being deficient remain poorly understood. Comparing root exudates from axenically grown maize plants exposed to N, K, P, or Fe deficiency showed a higher release of glutamate, glucose, ribitol, and citrate from Fe‐deficient plants, while P deficiency stimulated the release of γ‐aminobutyric acid and carbohydrates. Potassium‐starved plants released less sugars, in particular glycerol, ribitol, fructose, and maltose, while under N deficiency lower amounts of amino acids were found in root exudates. Principal‐component analysis revealed a clear separation in the variation of the root‐exudate composition between Fe or P deficiency versus N or K deficiency in the first principal component, which explained 46% of the variation in the data. In addition, a negative correlation was found between the amounts of sugars, organic and amino acids released under deficiency of a certain nutrient and the diffusion coefficient of the respective nutrient in soils. We thus hypothesize that the release of dominant root exudates such as sugars, amino acids, and organic acids by roots may reflect an ancient strategy to cope with limiting nutrient supply.     

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.     

Iron deficiency stress response of various c‐3 and c‐4 grain crop genotypes: Strategy II mechanism evaluated

The relative amount of phytosiderophore produced by various Strategy II plants has been categorized as follows: barley (Hordeum vulgare L.) > wheat (Triticum aestivum L.) > oat (Avena byzantina C. Koch.) > rye (Secale cereale L.) >> corn (Zea mays L.) >> sorghum (Sorghum bicolor (L.) Moench) > rice (Oryza sativa L.). With the exception of rice, these plants developed under oxidized soil conditions, and the C‐3 species produce more phytosiderophore than C‐4 species under Fe‐deficiency stress. Iron‐efficient Coker 227 oat produced phytosiderophore in response to Fe‐deficiency stress, while Fe‐inefficient TAM 0–312 oat did not. Although Fe‐efficient WF9 corn and Fe‐inefficient ys₁ corn differed in their ability to obtain Fe, neither produced sufficient quantities of phytosiderophore to explain these differences. The objectives of this research were to determine: (a) if phytosiderophore production of Fe‐deficiency stressed C‐4 species millet (Panicum miliaceum L.) and corn is low or absent compared to identically stressed C‐3 species oat and barley, and (b) if native, inbred and hybrid corn cultivars differ in ability to produce and utilize phytosiderophores.

Although release of phytosiderophore for Fe‐stressed corn and millet was generally lower than oat, quantity of release was not always related to obtaining Fe and maintaining green plants. Barley maintained high leaf Fe and low chlorosis with a minor release of phytosiderophore. Oat with increased release acted similarly to barley, whereas a relatively high release of phytosiderophore from White maize did not effect Fe uptake or greening. Likewise, small amounts of phytosiderophore were produced by all corn types, but corn was generally unable to obtain adequate Fe from the growth medium. Two of the native corns, Coneso and Tepecintle, maintained relatively low chlorosis, but they differed in phytosiderophore release. Thus, it appears that the C‐4 plants studied herein generally release a lower amount of phytosiderophore than do C‐3 species, but overcoming Fe‐deficiency chlorosis is not guaranteed by such release. The Strategy II mechanism of mere release of phytosiderophore and consequential Fe acquisition appears simplistic. There is a need for understanding what other factors are involved.     

Iron deficiency stress responses of five gram‐inaceous monocots

Plant growth, leaf chlorosis, root reductive capacity, rhizosphere pH, and phytosiderophore release capacity were used as indices to evaluate the responses of maize (Zea mays L. cv ‘clipper'), millet (Pennisetum glaucum L. cv. ‘Dwarf Gero'), sorghum (Sorghum bicolor L. cv. YG 5760), barnyard grass (Echinochloa crus galli L. cv: unknown), wheat (Triticum aestivum L. cv. ‘tonic'), and white lupin (Lupinus albus L. cv ‘lucky') to iron‐deficiency stress. Generally, root and shoot dry matter increased with iron treatment and leaves became less chlorotic. Neither the order nor the magnitude of the root reductive capacities of the monocots studied was affected by iron deprivation, but these reductive capacities and the changes in rhizosphere pH differed markedly. Significant iron stress‐induced phytosiderophore release was observed only in wheat and sorghum in which accompanying increases in rhizosphere pH were also evident. Such phytosiderophore release matched the severity of leaf chlorosis and iron uptake and depended on the form in which the element was supplied. These results, from experiments conducted in non‐axenic hydroponic cultures, indicate that in iron‐ deficiency stress mechanisms ‐ similar to those found in dicots ‐could account for iron uptake in some graminaceous monocots, and that strategy II‐type response proposed for all in this category of plants would be an over simplification.     

Iron-Deficiency Responses in Cereal Seedlings

Acidity release as a characteristic of iron (Fe) deficiency stress responses was investigated during early seedling establishment of maize (Zea mays L.), barley (Hordeum vulgare L.), winter wheat (Triticum aestivum L.), and winter rye (Secale cereale L.). Ferric reduction as another characteristic of Fe deficiency stress responses was also investigated in maize. This has previously been considered a process only occurring in iron-stressed dicotyledonous plants. Measurements were made of the Fe efflux from maize endosperm under Fe-deficient and non-limiting conditions. The scutellum of Fe-stressed cereal seedlings (all species investigated) increased agar acidification by 30–40%, while manganese (Mn) or zinc (Zn) stressed seedlings did not demonstrate this reaction. Among the maize genotypes studied (‘VIR-7569’ and ‘VIR-7882’), the Fe-stressed acid release was more prolonged in relatively less Fe-efficient cv. ‘VIR-7569.’ In maize, the induced enhancement of acid release was accompanied by an increase in Fe efflux from endosperm. Conversely, maize scutellum Fe^{3 +}-reductase activity was not deficiency-dependent on Fe, indicating that enhanced acidity release is a biological phenomenon facilitating Fe supply at early development of graminaceous plants. In cereal seedlings, the coexistence of two marker responses to Fe-deficiency (Strategy I and Strategy II), which are usually typical of different taxonomic groups (dicotyledonous and graminaceous plants) can be regarded as evidence in favour of the hypothesis of common genesis in dicotyledonous and graminaceous plants.     

Root-induced changes in the rhizosphere: Importance for the mineral nutrition of plants

Root-induced changes in the rhizosphere may affect mineral nutrition of plants in various ways. Examples for this are changes in rhizosphere pH in response to the source of nitrogen (NH₄-N versus NO₃-N), and iron and phosphorus deficiency. These pH changes can readily be demonstrated by infiltration of the soil with agar containing a pH indicator. The rhizosphere pH may be as much as 2 units higher or lower than the pH of the bulk soil. Also along the roots distinct differences in rhizosphere pH exist. In response to iron deficiency most plant species in their apical root zones increase the rate of H⁺ net excretion (acidification), the reducing capacity, the rate of Fe^III reduction and iron uptake. Also manganese reduction and uptake is increased several-fold, leading to high manganese concentrations in iron deficient plants. Low-molecular-weight root exudates may enhance mobilization of mineral nutrients in the rhizosphere. In response to iron deficiency, roots of grass species release non-proteinogenic amino acids (?phytosiderophores”?) which dissolve inorganic iron compounds by chelation of Fe^III and also mediate the plasma membrane transport of this chelated iron into the roots. A particular mechanism of mobilization of phosphorus in the rhizosphere exists in white lupin (Lupinus albus L.). In this species, phosphorus deficiency induces the formation of so-called proteoid roots. In these root zones sparingly soluble iron and aluminium phosphates are mobilized by the exudation of chelating substances (probably citrate), net excretion of H⁺ and increase in the reducing capacity. In mixed culture with white lupin, phosphorus uptake per unit root length of wheat (Triticum aestivum L.) plants from a soil low in available P is increased, indicating that wheat can take up phosphorus mobilized in the proteoid root zones of lupin. At the rhizoplane and in the root (root homogenates) of several plant species grown in different soils, of the total number of bacteria less than 1 % are N₂-fixing (diazotrophe) bacteria, mainly Enterobacter and Klebsiella. The proportion of the diazotroph bacteria is higher in the rhizosphere soil. This discrimination of diazotroph bacteria in the rhizosphere is increased with foliar application of combined nitrogen. Inoculation with the diazotroph bacteria Azospirillum increases root length and enhances formation of lateral roots and root hairs similarly as does application of auxin (IAA). Thus rhizosphere bacteria such as Azospirillum may affect mineral nutrition and plant growth indirectly rather than by supply of nitrogen.     

Phytosiderophore release as a criterion for genotypic evaluation of iron efficiency in oat

Iron (Fe) deficiency in small grains grown on calcareous soils results in reduced yields, is difficult and expensive to treat with fertilizer, and is complicated to overcome by genetic field screening due to heterogeneous soil and environmental conditions. Recently, phytosiderophore release has been linked to ability of species and genotypes to resist Fe‐deficiency chlorosis. We propose a laboratory technique to measure phytosiderophore release by Fe‐deficient oat (Avena sativa L.) genotypes as a selection method for Fe‐deficiency chlorosis resistance in oat. Plants were grown in Fe‐limiting nutrient solution and phytosiderophore release was measured on 11 days. Summations of daily phytosiderophore release by 17 oat genotypes correlate well with Fe‐deficiency chlorosis scores in the field (r = ‐0.70, p = 0.01). The proposed method consistently identified the genotypes most susceptible to Fe deficiency but did not clearly separate the moderately susceptible genotypes. In these latter genotypes, other factors such as active uptake sites, root growth rate, utilization of acquired Fe, or soil interactions may be modifying factors to phytosiderophore in Fe efficiency. Quantification of phytosiderophore provides a useful selection criterion for oat by eliminating the most inefficient types and with refinement, may become a powerful tool for identifying Fe efficiency in grass crops.     

Responses by iron-efficient and inefficient oat cultivars to inoculation with siderophore-producing bacteria in a calcareous soil

Rhizosphere bacteria may enhance plant uptake of Fe by producing siderophores that chelate sparingly soluble Fe³⁺ in calcareous soils. To evaluate the extent to which plants benefit from colonization of the roots by prolific siderophore-producing bacteria, we inoculated two oat cultivars with six strains of bacteria that produced high concentrations of siderophores under Felimiting conditions in vitro. Oat cv Coker 227, an Fe-efficient cultivar, which produces the phytosiderophore avenic acid, and cv TAM 0-312, and Fe-inefficient cultivar, which does not produce the phytosiderophore, were grown in a calcareous soil (Weswood silt loam) on a light bench in the laboratory. Half of the plants were fertilized with a nutrient solution containing 5 mM Fe and half with a nutrient solution containing no Fe. After 6 weeks of growth, we compared colonization of the roots by the inoculant bacteria and the dry weight and Fe content of roots and shoots. Three species of Pseudomonas colonized the roots of both oat cultivars in high numbers (10⁶ cells g^-1 root dry weight), whereas the remaining bacteria colonized the roots in substantially lower numbers (10⁴ cells g^-1 root dry weight). Plants fertilized with 5 mM Fe were larger and supported greater numbers or rhizosphere bacteria per gram of root than plants not supplied with Fe. Comparisons of the Fe content and dry weight of roots and shoots revealed few significant differences between inoculated and uninoculated plants, or among the plants inoculated with the different strains of siderophore-producing bacteria. The differences that were observed revealed no consistent response to inoculation. We conclude that inoculation of the roots of the two oat cultivars with bacteria that produce high concentrations of siderophores in response to an Fe deficiency had little or no effect on Fe acquisition by the plants.