Changes in expression of proteins involved in alleviation of Fe-deficiency by sulfur nutrition in Brassica napus L.

In order to characterize the significance of sulfur (S) nutrition in protein expression under iron (Fe)-deficient conditions, gel-based proteomic analysis was performed with the leaves of Brassica napus exposed to S and Fe combined treatments: sufficient in S and Fe (+S/+Fe, control), sufficient S but Fe deprived (+S/?Fe), deprived S but sufficient Fe (?S/+Fe), and deprived S and Fe (?S/?Fe). The resulting data showed that 15 proteins were down-regulated due to production of oxidative damage as indicated by H₂O₂ and O ₂ ^?1 localizations and due to leaf chlorosis in leaves in S-deprived leaves either in presence (?S/+Fe) or absence of Fe (?S/?Fe), whereas these down-regulated proteins were well expressed in the presence of S (+S/?Fe) compared to control (+S/+Fe). In addition, two proteins were up-regulated under S-deprived condition in presence (?S/+Fe) and absence of (?S/?Fe) Fe. The functional classification of these identified proteins was estimated that 40 % of the proteins belong to chloroplast precursor, and rest of the proteins belongs to hypothetical proteins, RNA binding, secondary metabolism and unknown proteins. On the other hand, five protein spots from S deprived (?S/+Fe) and ten spots from Fe deprived (?S/?Fe) conditions were absent, whereas they were well expressed in presence of S (+S/?Fe) compared to control plants (+S/+Fe). These results suggest that sulfur nutrition plays an important role in alleviating protein damage in Fe-deficient plants and adaptation to Fe-deficiency in oilseed rape.     

Effect of cadmium on iron uptake in cucumber roots: A Mössbauer-spectroscopic study

The uptake and accumulation of iron in cucumber roots exposed to cadmium were investigated with Fe sufficient and deficient cucumber plants using Mössbauer spectroscopy, Inductively Coupled Plasma Mass Spectrometry (ICP-MS) and ferric chelate reductase activity measurements. Both Fe sufficient and Fe deficient plants were applied. In the case of Fe sufficient cucumber roots grown in nutrient solution with 10 μM Cd no changes were found in the occurrence of Fe species (mostly hydrous ferric oxides and ferric-carboxylate complexes) compared to the control where no Cd was added. In the Fe deficient roots pretreated with 0, 0.1, 1, 10 and 100 μM Cd for 3 h then supplied also with 0.5 mM ⁵⁷Fe-citrate for 30 min, Fe^II was identified in a hexaaqua complex form. The relative amount of Fe^II was decreasing simultaneously with increasing Cd concentration, while the relative occurrence of Fe^III species and total Fe concentration were increasing. The results support the inhibitory effect of Cd on Fe-chelate reduction. Although the reductase activity at 10 and 100 μM Cd treatment was lower than in the iron sufficient control plants, Fe^II could be identified by Mössbauer spectroscopy whereas in the Fe sufficient control, this form was below detection limit. These data demonstrate that the influx and the reoxidation of Fe^II was decreased by Cd, consequently, they refer to the competition of Cd²⁺ and Fe²⁺ during the membrane transport and the inhibition of the reoxidation process.     

Comparison of cadmium-induced iron-deficiency responses and genuine iron-deficiency responses in Malus xiaojinensis

The effects of the heavy metal Cd in Malus xiaojinensis were investigated using hydroponic cultures. Chlorophyll and Fe concentrations in young leaves were markedly decreased by Cd treatment, although Fe concentration was significantly enhanced in the roots. A comparative examination of the Fe-deficiency responses due to Fe deficiency and Cd treatment was also performed. Both Fe deficiency and Cd treatment induced responses similar to those of Fe-deficiency in M. xiaojinensis, including acidification of the rhizosphere, enhanced Fe(III) chelate reductase activity, and upregulation of the Fe-deficiency-responsive genes MxIRT1 and MxFRO2-Like. However, the Fe-deficiency responses induced by Cd treatment were different in intensity and timing from those induced by Fe deficiency.     

Does Iron Deficiency in Pisum sativum Enhance the Activity of the Root Plasmalemma Iron Transport Protein?

Roots of Fe-sufficient and Fe-Deficient pea (Pisum sativum L.) were studied to determine the effect of Fe-deficiency on the activity of the root-cell plasmalemma Fe²⁺ transport protein. Rates of Fe(III) reduction and short-term Fe²⁺ influx were sequentially determined in excised primary lateral roots using Fe(III)-ethylene-diaminetetraacetic acid (Fe[III]-EDTA). Since the extracellular Fe²⁺ for membrane transport was generated by root Fe(III) reduction, rates of Fe²⁺ influx for each root system were normalized on the basis of Fe(III) reducing activity. Ratios of Fe²⁺ influx to Fe(III) reduction (micromole Fe²⁺ absorbed/micromole Fe[III] reduced) revealed no enhanced Fe²⁺ transport capacity in roots of Fe-deficient peas (from the parental genotype, Sparkle) or the functional Fe-deficiency pea mutant, E107 (derived from Sparkle), relative to roots of Fe-sufficient Sparkle plants. Data from studies using 30 to 100 micromolar Fe(III)-EDTA indicated a linear relationship between Fe²⁺ influx and Fe(III) reduction (Fe²⁺ generation), while Fe²⁺ influx saturated at higher concentrations of Fe(III)-EDTA. Estimations based on current data suggest the Fe²⁺ transport protein may saturate in the range of 10^−4.8 to 10⁻⁴ molar Fe²⁺. These results imply that for peas, the physiological rate limitation to Fe acquisition in most well-aerated soils would be the root system's ability to reduce soluble Fe(III)-compounds.     

The influence of redox conditions in soils on extraplasmatic Fe-loading of plant roots

Loading of extraplasmatic Fe, as a potential storage pool for Fe nutrition, was studied in roots of maize grown under hypoxic conditions in soil culture. Extraplasmatic Fe loading was investigated depending on (i) duration of flooding (0, 1, 2 or 4 days) and (ii) microbial activity as affected by graduated addition of carbon sources (0, 2 or 10 g each starch and cellulose kg^?1 soil). Maize plants were grown in a soil culture system with root systems enclosed in membrane bags to avoid Fe contamination of the root surface by soil particles. Due to the high redox buffer capacity of the Haplic Luvisol employed for the experiments, flooding treatments induced only moderately reducing conditions (～?300 mV) and a slight increase of extraplasmatic Fe loading (41\to165 mg kg^?1 d.m.). Strongly reducing conditions (?100 mV) associated with a high Fe²⁺ concentration in the soil solution and a significant increase of extraplasmatic Fe (1190 mg kg^?1 d.m.) were obtained only after application of high amounts of organic carbon (10 g starch and 10 g cellulose kg^?1 soil), which accompanied by unrealistic reducing conditions due to intense stimulation of microbial growth. The expression of effects only under extremely high application level of organic carbon (～?33 t C ha^?1) suggest that similar to aerobic conditions, extraplasmatic Fe-loading under transient hypoxia is probably of limited ecological significance for the iron nutrition of higher plants, at least in soils with a high redox buffer capacity as employed in the present study. Abbreviations: DHA – dehydrogenase activity; d.m. – dry matter; DOC – dissolved organic carbon; Eh – redox potential; PIXE – proton-induced X-ray emission; STIM – scanning transmission ion microscopy.     

Effectiveness of Ethylenediamine-N(o-hydroxyphenylacetic)-N′(p-hydroxyphenylacetic) acid (o,p-EDDHA) to Supply Iron to Plants

The Fe chelate o,p-EDDHA/Fe³⁺, in addition to o,o-EDDHA/Fe³⁺, was found recently to be a component of commercial EDDHA/Fe³⁺ chelates. The European Regulation on fertilisers has included o,p-EDDHA as an authorized chelating agent. The efficacy of o,o-EDDHA/Fe³⁺, o,p-EDDHA/Fe³⁺ and EDTA/Fe³⁺ chelates as Fe sources in plant nutrition was studied. Iron-chelate reductase (FC-R) in young cucumber plants (Cucumis sativus L.) roots reduced o,p-EDDHA/Fe³⁺ faster than o,o-EDDHA/Fe³⁺, EDTA/Fe³⁺ and a commercial source of EDDHA/Fe³⁺. The o,p-EDDHA/Fe³⁺ chelate was also more effective than the o,o-EDDHA/Fe³⁺ in decreasing the severity of Fe-deficiency chlorosis in leaves of young soybean (Glycine max L.) plants grown hydroponically. The o,p-EDDHA ligand was more effective in the short-term than the EDTA and o,o-EDDHA ligands at dissolving Fe from selected Fe minerals and soils. However, the ultimate quantity of dissolve Fe was greatest with the o,o-EDDHA ligand.     

Investigation of iron pools in cucumber roots by Mössbauer spectroscopy: direct evidence for the Strategy I iron uptake mechanism

Distinct chemical species of iron were investigated by Mössbauer spectroscopy during iron uptake into cucumber roots grown in unbuffered nutrient solution with or without ⁵⁷Fe-citrate. Mössbauer spectra of iron deficient roots supplied with 10–500 μM ⁵⁷Fe-citrate for 30–180 min and 24 h and iron-sufficient ones, were recorded. The roots were analysed for Fe concentration and Fe reductase activity. The Mössbauer parameters in the case of iron-sufficient roots revealed high-spin iron(III) components suggesting the presence of Fe^III-carboxylate complexes, hydrous ferric oxides and sulfate–hydroxide containing species. No Fe^II was detected in these roots. However, iron-deficient roots supplied with 0.5 mM ⁵⁷Fe^III-citrate for 30 min contained significant amount of Fe^II in a hexaaqua complex form. This is a direct evidence for the Strategy I iron uptake mechanism. Correlation was found between the decrease in Fe reductase activity and the ratio of Fe^II–Fe^III components as the time of iron supply was increased. The data may refer to a higher iron reduction rate as compared to its uptake/reoxidation in the cytoplasm in accordance with the increased reduction rate in iron deficient Strategy I plants.     

AhDMT1, a Fe<Superscript>2+</Superscript> transporter,is involved in improving iron nutrition and N<Subscript>2</Subscript> fixation in nodules of peanut intercropped with maize in calcareous soils

Peanut (Arachis hypogaea L.) is an important legume providing edible proteins and N₂ fixation. However, iron deficiency severely reduces peanut growth in calcareous soils. The maize/peanut intercropping effectively improves iron nutrition and N₂ fixation of peanut under pot and field conditions on calcareous soils. However, little was known of how intercropping regulates iron transporters in peanut. We identified AhDMT1 as a Fe²⁺ transporter which was highly expressed in mature nodules with stronger N₂ fixation capacity. Promoter expression analysis indicated that AhDMT1 was localized in the vascular tissues of both roots and nodules in peanut. Short-term Fe-deficiency temporarily induced an AhDmt1 expression in mature nodules in contrast to roots. However, analysis of the correlation between the complex regulation pattern of AhDmt1 expression and iron nutrition status indicated that sufficient iron supply for long term was a prerequisite for keeping AhDmt1 at a high expression level in both, peanut roots and mature nodules. The AhDmt1 expression in peanut intercropped with maize under 3 years greenhouse experiments was similar to that of peanut supplied with sufficient iron in laboratory experiments. Thus, the positive interspecific effect of intercropping may supply sufficient iron to enhance the expression of AhDmt1 in peanut roots and mature nodules to improve the iron nutrition and N₂ fixation in nodules. This study may also serve as a paradigm in which functionally important genes and their ecological significance in intercropping were characterized using a candidate gene approach.     

Fe-deficiency stress response in Fe-deficiency resistant and susceptible subterranean clover: importance of induced H+ release

Plants can exhibit Fe-deficiency stress response when they areexposed to Fe-deficiency conditions. The relative importanceof the individual Fe-deficiency stress-response reactions, forexample, increased release of H⁺ from roots, enhanced root plasmamembrane-bound Fe³⁺ -reductase activity, and release of reductant,in Fe-deficiency resistance is not understood. To address thisproblem, the Fe-deficiency stress response of two cultivarsof subterranean clover (subclover), Koala (Trifolium brachycalycinumKatzn. and Morley) (Fe-deficiency resistant) and Karridale (T.subterraneum L.) (Fe-deficiency susceptible), were evaluated.The plants were cultured hydroponically at 0 (–Fe) and30 (+Fe) µM Fe³⁺ EDTA conditions. After 6 d Fe treatment,the –Fe Koala and Karridale decreased the pH of the nutrientsolution by 1.83 and 0.79 units, respectively, while the +Feplants increased the pH of the nutrient solution. The H⁺ -releaserate of the –Fe Koala determined 7 d after Fe treatmentinitiation was more than three times higher than that of the–Fe Karridale. The –Fe plants had a significantlyenhanced Fe³⁺ -reduction rate compared with the +Fe plants foreach cultivar, but the resistant cultivar did not exhibit ahigher root Fe³⁺ -reduction rate than the susceptible cultivarat each Fe treatment. Reductant release from the roots of subcloverwas negligible. These results indicate that Fe-deficiency-inducedH⁺ release may be the predominant factor influencing Fe-deficiencyresistance in subclover. Key words: Fe-deficiency, Fe³⁺ reduction, H⁺ release, stress response, Trifolium     

The effect of sodium bicarbonate on plant performance and iron acquisition system of FA-5 (Forner-Alcaide 5) citrus seedlings

This work studies the effect of bicarbonate on plant performance and the iron acquisition system of Forner-Alcaide 5 (FA-5) seedlings, a citrus genotype known for its tolerance to calcareous soils. Plants were irrigated for 6 weeks with or without 10 mM NaHCO₃. Treatment significantly decreased shoot growth, photosynthetic levels and iron concentration in shoots and roots. o,o-⁵⁷FeEDDHA experiments indicated that ⁵⁷Fe uptake by roots was inhibited in treated plants. Moreover, those seedlings accumulated more ⁵⁷Fe in roots, and enhanced mRNA accumulation of ferric reductase genes FRO1 and FRO2 and FC-R activity in roots. H⁺-ATPase activity and HA1 gene expression were also increased, while HA2 was not affected. In addition, expression of the iron transporter gene IRT1 was increased, while IRT2 was not significantly affected. Finally, according to PEPC enzymatic activity, PEPC1 gene expression was higher in treated roots. In conclusion, it appears that bicarbonate prevents medium acidification by roots, thus reducing Fe²⁺ uptake. Accordingly, Fe deficiency enhanced the expression of some genes related with the Fe acquisition system (IRT1, FRO1, FRO2, HA1 and PEPC1) and the activity of the corresponding enzymes, which appear to constitute an adaptive mechanism of FA-5 in these soils.     

In-vitro-cultured subclover root can develop Fe-deficiency stress response

The Fe-deficiency stress response is induced in most plants under Fe-deficient conditions, but whether the shoot and/or the root control development of the stress response is not known. The objectives of the present study were to determine whether in-vitro-cultured subclover roots can develop Fe-deficiency stress response and to examine this approach as a possible screening technique for Fe-deficiency resistance. One-cm long root tips of subclover seedlings were cultured in modified White's medium without (-Fe) or with (+Fe) 100 μM Fe³⁺EDTA. Root Fe³⁺ reduction and H⁺ release were evaluated. On the first day after transfer to the -Fe medium, the Fe-deficiency-resistant cultivar Koala (Trifolium brachycalycinum Katzn. and Morley) started to release H⁺, resulting in a decrease in pH of the culture medium, while the susceptible cultivar Karridale (T. subterraneum L.) did not release H⁺ until the second day. The H⁺-release rate of the -Fe Koala was approximately twice as high as that of the -Fe Karridale for the first 4 days of -Fe treatment. Both Koala and Karridale reached their highest H⁺-release rates on the fourth day after -Fe treatment initiation. The +Fe Koala released H⁺ after several days of culture, but the H⁺ release of the -Fe Koala was severalfold greater than that of the +Fe Koala. The implicit correlation between H⁺ release and Fe-deficiency resistance was substantiated by using a series of subclover cultivars with a range of susceptibilities to Fe deficiency. The pH of the -Fe culture media of the series of cultivars was positively correlated to their Fe-chlorosis scores reported in previous research. The results of the present study indicate that root itself has the full ability to develop Fe-deficiency stress response and the response is dependent on the root Fe status. The results also suggest that root culture could be used as a simple and efficient alternative technique for screening germplasm for Fe-deficiency resistance.     

Photosynthetic physiological performance and proteomic profiling of the oleaginous algae <Emphasis Type="Italic">Scenedesmus acuminatus</Emphasis> reveal the mechanism of lipid accumulation under low and high nitrogen supplies

In this study, we presented cellular morphological changes, time-resolved biochemical composition, photosynthetic performance and proteomic profiling to capture the photosynthetic physiological response of Scenedesmus acuminatus under low nitrogen (3.6 mM NaNO₃, N^?) and high nitrogen supplies (18.0 mM NaNO₃, N⁺). S. acuminatus cells showed extensive lipid accumulation (53.7% of dry weight) and were enriched in long-chain fatty acids (C16 & C18) under low nitrogen supply. The activity of PSII and photosynthetic rate decreases, whereas non-photochemical quenching and dark respiration rates were increased in the N^? group. In addition, the results indicated a redistribution of light excitation energy between PSII and PSI in S. acuminatus exists before lipid accumulation. The iTRAQ results showed that, under high nitrogen supply, protein abundance of the chlorophyll biosynthesis, the Calvin cycle and ribosomal proteins decreased in S. acuminatus. In contrast, proteins associated with the photosynthetic machinery, except for F-type ATPase, were increased in the N⁺ group (N⁺, 3 vs. 9 days and 3 days, N⁺ vs. N^?). Under low nitrogen supply, proteins involved in central carbon metabolism, fatty acid synthesis and branched-chain amino acid metabolism were increased, whereas the abundance of proteins of the photosynthetic machinery had decreased, with exception of PSI (N^?, 3 vs. 9 days and 9 days, N⁺ vs. N^?). Collectively, the current study has provided a basis for the metabolic engineering of S. acuminatus for biofuel production.     

pH Changes Associated with Iron-Stress Response

When Fe-inefficient T3238fer and Fe-efficient T3238FER tomatoes were supplied iron, and nitrogen as nitrate, they increased the pH of the nutrient culture. When they were supplied nitrogen as ammonium, they decreased the pH. When Fe supply was limited, Fe-stress response developed in T3238FER that opposed the usual nitrate response and decreased, rather than increased, the pH. A “reductant” which reduced Fe³⁺ to Fe²⁺ was released from the roots of these plants and lowered the pH; and there was a tremendous increase in the uptake of Fe. T3238fer did not produce “reductant” in response to Fe-stress; the pH increased, and the plants developed Fe-deficiency when nitrogen was supplied as nitrate. Nitrogen nutrition and iron-stress response are important factors associated with iron chlorosis in plants. Release of hydrogen ions from roots of Fe-stressed plants is caused by more than response to imbalanced uptake of cations and anions.     

5-Aminolevulinic acid improves photosynthetic gas exchange capacity and ion uptake under salinity stress in oilseed rape (Brassica napus L.)

Salinity is one of the major constraints in oilseed rape (Brassica napus L.) production. One of the means to overcome this constraint is the use of plant growth regulators to induce plant tolerance. To study the plant response to salinity in combination with a growth regulator, 5-aminolevulinic acid (ALA), oilseed rape plants were grown hydroponically in greenhouse conditions under three levels of salinity (0, 100, and 200 mM NaCl) and foliar application of ALA (30 mg/l). Salinity depressed the growth of shoots and roots, and decreased leaf water potential and chlorophyll concentration. Addition of ALA partially improved the growth of shoots and roots, and increased the leaf chlorophyll concentrations of stressed plants. Foliar application of ALA also maintained leaf water potential of plants growing in 100 mM salinity at the same level as that of the control plants, and there was also an improvement in the water relations of ALA-treated plants growing in 200 mM. Net photosynthetic rate and gas exchange parameters were also reduced significantly with increasing salinity; these effects were partially reversed upon foliar application with ALA. Sodium accumulation increased with increasing NaCl concentration which induced a complex response in the macro-and micronutrients uptake and accumulation in both roots and leaves. Generally, analyses of macro- (N, P, K, S, Ca, and Mg) and micronutrients (Mn, Zn, Fe, and Cu) showed no increased accumulation of these ions in the leaves and roots (on dry weight basis) under increasing salinity except for zinc (Zn). Foliar application of ALA enhanced the concentrations of all nutrients other than Mn and Cu. These results suggest that under short-term salinity-induced stress (10 days), exogenous application of ALA helped the plants improve growth, photosynthetic gas exchange capacity, water potential, chlorophyll content, and mineral nutrition by manipulating the uptake of Na⁺.     

Accumulation and within-seed distribution of iron in common bean and soybean

The influence of times of applying FeEDDHA on seed yield and Fe accumulation by four common bean (Phaseolus vulgaris L.) and two soybean (Glycine max L.) genotypes grown on a calcareous soil was studied under greenhouse conditions. The soybean genotypes, unlike the common bean genotypes, developed Fe-deficiency chlorosis and responded to application of the chelate. A preplant application of FeEDDHA was more efficacious than a flowering application in increasing seed yield of soybean. In contrast, the flowering application was much more effective than the preplant application for increasing seed Fe concentration [Fe] of both species. Percentage of seed Fe located in the seed coat of the common bean genotypes ranged from approximately 5 to 40% and was little affected by FeEDDHA. This within-seed distribution of Fe was correlated with methanol-extractable seed-coat pigments absorbing at 500 nm, presumably anthocyanins, but not with condensed tannins (proanthocyanidins). The soybean genotypes did not accumulate anthocyanins or tannins in the seed coat. Seed of Fe-deficient soybean plants without FeEDDHA had appreciably lower [Fe] and had a lower percentage of seed Fe in the seed coat than treated plants. Within-seed distribution of Fe should be considered in plant breeding because of concerns about both human nutrition and early seedling growth. Abbreviations: DTPA – diethylenetrinitrilopentaacetic acid; EDDHA – ethylenediamine di(o-hydroxyphenylacetic acid) acid; SPAD – single photon avalanche diode     

Effectiveness of N,N′-Bis(2-hydroxy-5-methylbenzyl) ethylenediamine-N,N′-diacetic acid (HJB) to supply iron to dicot plants

Iron chlorosis is commonly corrected by the application of EDDHA chelates, whose industrial synthesis produces o,oEDDHA together with a mixture of regioisomers and other unknown by-products. HJB, an o,oEDDHA analogous, is a new chelating agent with a purer synthesis pathway than EDDHA. The HJB/Fe³⁺ stability constant is intermediate between the racemic and meso o,oEDDHA/Fe³⁺ stereoisomers. This work studied the efficacy of HJB as a Fe source in plant nutrition. No significant differences between o,oEDDHA/Fe³⁺, HJB/Fe³⁺ and HBED/Fe³⁺ were observed when they are used as substrates of the iron-chelate reductase of mild chlorotic cucumber plants. Chelates prepared with the stable isotope ⁵⁷Fe were used in both soil and hydroponic experiments. In the hydroponic experiment, nutrient solutions with low doses of chelates were renewed weekly. Soybean plants treated with o,oEDDHA/⁵⁷Fe³⁺ recorded the highest results in biomass, SPAD index and Fe nutrition. In the soil experiment, chelates were added once at a rate of 2.5 mg Fe per kg of a calcareous soil. Soybean plants treated with HJB/⁵⁷Fe³⁺ recorded a higher biomass and SPAD index in young leaves than the plants treated with o,oEDDHA/⁵⁷Fe³⁺; however, ⁵⁷Fe and total Fe concentrations in leaves were lower. The results of both pot experiments are associated with a faster ability by o,oEDDHA to provide Fe to the plants and with a more continuous supply of Fe from HJB/Fe³⁺. HJB/⁵⁷Fe³⁺ effectively alleviated the Fe-deficiency chlorosis of soybean with a longer lasting effect than o,oEDDHA/⁵⁷Fe³⁺.     

Enzymatic responses of cucumber roots to different levels of Fe supply

Development of the coordinated response to decreasing Fe availability was studied in cucumber plants grown in nutrient solution (NS) over a range of Fe^III-EDTA concentrations (from 0.1 to 80 M). Physiological and biochemical parameters were evaluated in intact roots, root extracts and plasma membrane (pm) vesicles. Acidification of the NS was evident in plants grown at 1 M Fe^III-EDTA and inversely related to the external Fe concentration. Fe^III-EDTA reduction by intact roots was also gradually depressed by increasing Fe supply. The rate of net nitrate uptake by the roots was directly related to the amount of Fe^III-EDTA added to the NS. Activity of pmH⁺-ATPase was significantly higher in plants grown without added Fe as compared to those grown at 80 M Fe. A lower increase, dependent on Fe concentration, was observed at 0.1, 1, 5 or 10 M Fe^III-EDTA. Activity of pmFe^III-EDTA reductase was also increased by Fe deprivation and strongly correlated with pmH⁺-ATPase activity. PEP-carboxylase activity gradually increased with decreasing Fe concentration in the NS. Changes in activity and amount of the enzyme showed a close correlation with parameters measured in intact roots (nitrate uptake, Fe^III-EDTA reduction). Results show that the development of the Fe-deficiency response in cucumber roots can be finely tuned by the level of Fe supply. Adjustments to different levels of available Fe involve a correlated modulation of pm-associated enzymes. PEP-carboxylase activity appeared to be a suitable metabolic marker of the Fe nutritional status of the plant.     

Sulfur deprivation and nitrogen metabolism in maize seedlings

The objective of this experiment was to elucidate the manner in which N metabolism is influenced by S nutrition. Maize (Zea mays L.) seedlings supplied with Hoagland solution minus SO₄²⁻ exhibited S deficiency symptoms 12 days after emergence. Prior to development of these symptoms, a decline in leaf blade nitrate reductase (NR, EC 1.6.6.1) activity was observed in S-deprived seedlings compared to normal seedlings. Twelve days after emergence, in vitro NR activity was diminished 50% compared to normal seedlings. Glutamine synthetase (EC 6.3.1.2) and NAD-glutamate dehydrogenase (EC 1.4.1.2) activities were less severely affected (19 and 13%, respectively, at day 12). NADP-glutamate dehydrogenase (EC 1.4.1.4) activity and leaf blade fresh weight were not altered by S deprivation. Concentrations of soluble protein and chlorophyll (a and b) in leaf blades were reduced 18 and 25%, respectively, at day 12. A significantly higher concentration of NO₃⁻-N was observed for leaf blade and stem (culms, leaf sheaths, and unfurled leaves) fractions (46 and 31%, respectively) in S-deprived plants. In contrast to the other parameters measured, NR activity in S-deprived seedlings could be readily restored to the normal level by addition of SO₄²⁻. The apparent preferential effect of S deprivation on NR activity could be causally related to the observed changes in NO₃⁻-N and soluble protein concentration.     

Neutrophilic, nitrate-dependent, Fe(II) oxidation by a Dechloromonas species

A species of Dechloromonas, strain UWNR4, was isolated from a nitrate-reducing, enrichment culture obtained from Wisconsin River (USA) sediments. This strain was characterized for anaerobic oxidation of both aqueous and chelated Fe(II) coupled to nitrate reduction at circumneutral pH. Dechloromonas sp. UWNR4 was incubated in anoxic batch reactors in a defined medium containing 4.5–5 mM NO₃ ^?, 6 mM Fe²⁺ and 1–1.8 mM acetate. Strain UWNR4 efficiently oxidized Fe²⁺ with 90 % oxidation of Fe²⁺ after 3 days of incubation. However, oxidation of Fe²⁺ resulted in Fe(III)-hydroxide-encrusted cells and loss of metabolic activity, suggested by inability of the cells to utilize further additions of acetate. In similar experiments with chelated iron (Fe(II)-EDTA), encrusted cells were not produced and further additions of acetate and Fe(II)-EDTA could be oxidized. Although members of the genus Dechloromonas are primarily known as perchlorate and nitrate reducers, our findings suggest that some species could be members of microbial communities influencing iron redox cycling in anoxic, freshwater sediments. Our work using Fe(II)-EDTA also demonstrates that Fe(II) oxidation was microbially catalyzed rather than a result of abiotic oxidation by biogenic NO₂ ^?.