Biosynthesis Of Phytosiderophores Research Articles

Comparative physiological, biochemical and transcriptomic analysis of hexaploid wheat (T. aestivum L.) roots and shoots identifies potential pathways and their molecular regulatory network during Fe and Zn starvation

The combined effect of iron (Fe) and zinc (Zn) starvation on their uptake and transportation and the molecular regulatory networks is poorly understood in wheat. To fill this gap, we performed a comprehensive physiological, biochemical and transcriptome analysis in two bread wheat genotypes, i.e. Narmada 195 and PBW 502, differing in inherent Fe and Zn content. Compared to PBW 502, Narmada 195 exhibited increased tolerance to Fe and Zn withdrawal by significantly modulating the critical physiological and biochemical parameters. We identified 25 core genes associated with four key pathways, i.e. methionine cycle, phytosiderophore biosynthesis, antioxidant and transport system, that exhibited significant up-regulation in both the genotypes with a maximum in Narmada 195. We also identified 26 microRNAs targeting 14 core genes across the four pathways. Together, core genes identified can serve as valuable resources for further functional research for genetic improvement of Fe and Zn content in wheat grain.

Identifying transcripts associated with efficient transport and accumulation of Fe and Zn in hexaploid wheat (T. aestivum L.)

Wheat (T. aestivum L.) is the second most important staple food crop consumed in the form of various end-use products across the world. However, it contains lower concentrations of Fe and Zn leading to micronutrient deficiency in human beings where wheat is the sole diet. Therefore, increasing grain Fe/Zn content in wheat has become priority in wheat breeding programmes across the world. Understanding the molecular mechanism of Fe/Zn transport and accumulation in grains is required to expedite the breeding process. For this purpose, whole seedling transcriptome analysis was conducted in four wheat genotypes (CRP 1660, Sonora 64, Vinata, : high, and DBW17: low) differing in grain Fe/Zn content under controlled and Fe/Zn deficient conditions. Twenty eight key transcripts involved in phytosiderophore biosynthesis, Fe/Zn uptake and transport were identified. Expression analysis of 12 of the transcripts using qPCR was conducted in seedling stage and flag leaf which exhibited greater differential accumulation in CRP 1660 followed by Vinata, Sonora 64 and DBW 17 in both flag leaf and seedling. However, there was significantly higher differential accumulation of the transcripts in flag leaf as compared to seedling. In CRP 1660, transcripts pertaining to phytosiderophore biosynthesis like DMAS1-B, NRAMP2 and NAAT2-D showed greater accumulation. Additionally, corresponding miRNAs were also identified for these 28 transcripts. The findings will help in better understanding of molecular basis of Fe/Zn transport and accumulation in grain and subsequent utilization in breeding to improve Fe/Zn content in wheat grain.

Ethylene regulation of root growth and phytosiderophore biosynthesis determines iron deficiency tolerance in wheat (Triticum spp)

Two separate experiments were conducted with four bread and two durum wheat cultivars, to test a hypothesis that high ethylene production will alter phytosiderophore production and release dynamics either directly, by competitive partitioning of SAM, or indirectly by causing changes in the root system architecture under iron deficiency. A significant decline in the leaf greenness index confirmed the Fe deficiency response across the wheat cultivars. Further, a significantly higher root to shoot ratio was measured for the durum wheat than the bread wheat cultivars under the iron deficiency treatment. Root attributes, although, were affected under iron deficiency, the variation between the bread and durum wheat cultivars were not too obvious. Durum wheat maintained a higher shoot, root and the whole plant iron levels than the bread wheat under Fe sufficient treatment while a role reversal was observed under Fe deficiency. Phytosiderophore production in the root tips and their release were induced under Fe deficiency and were higher for the bread than the durum wheat. A higher mRNA expression of ACS but a lower expression of NAS were identified with the durum wheat while the reverse was observed for the bread wheat. These results point towards a higher ethylene production and a lower phytosiderophore production in durum wheat as was also evidenced from observations on the ethylene release from the roots which was bettered in the presence of ethrel, an ethylene releasing chemical, but was completely inhibited in the presence of cobalt (Co), an ethylene synthesis inhibitor. An inhibited ethylene did improve the phytosiderophore production and phytosiderophore release for both the bread and the durum wheat cultivars under the iron deficiency treatment in the presence of cobalt. Since phytosiderophore production and release were not significantly altered over control, under Fe deficiency, in the presence of silver thiosulfate (STS), an ethylene action inhibitor, it could be concluded that ethylene does not affect NAS and that the effect of ethylene on the phytosiderophore production is direct and is caused by the dynamics of partitioning of S-adenosyl methionine (SAM) between the ethylene and the phytosiderophore biosynthetic pathways.

Uninhibited biosynthesis and release of phytosiderophores in the presence of heavy metal (HM) favors HM remediation.

Production of phytosiderophore (PS) has been causally related to iron-deficiency tolerance in cereals. However, PS can also chelate heavy metal and thus may represent a viable phytoextraction strategy on contaminated soils. Two separate experiments were conducted to assess the affect of heavy metal on phytosiderophore biosynthesis and their release in the rhizosphere of wheat. Root exudates were collected from 10-day-old wheat seedlings raised on Fe-deficient nutrient solution in the presence of 2.5, 5.0, and 10mM concentration of different heavy metals (Cd, Pb, and Ni) for 3-day period, for the phytosiderophore and the heavy metal analysis. Plant uptake of respective heavy metal was positively correlated with the heavy metal concentration of the nutrient solution. Phytosiderophore release was positively influenced in the presence of the heavy metal. Increasing concentration of Cd, Pb, and Ni showed positive correlation with the PS release until 5mM concentration followed by a decline at 10mM. However, a higher induction of PS release was measured in wheat seedlings treated with Cd and Pb than Ni. Further, transcript expression analysis of nicotianamine synthase (NAS) and nicotianamine amino transferase (NAAT), involved in phytosiderophore biosynthesis, was done in roots of 10-day-old Fe-deficient wheat subjected to 2.5, 5.0, and 10mM of Cd, Pb, and Ni. Both NAS and NAAT were expressed not only under Fe deficiency but also in the presence of Cd, Pb, and Ni. Sequencing of partial cDNA of NAS revealed a nucleotide length of 998bp, while multiple sequence alignment of NAS with HvNAS revealed 92% sequence similarity. This study irrevocably shows that phytosiderophore biosynthesis and release are not impaired in the presence of heavy metal and that phytosiderophore mediates the uptake of toxic heavy metal.

Transcriptome Analysis Reveals the Response of Iron Homeostasis to Early Feeding by Small Brown Planthopper in Rice.

It has been documented that planthopper attacks change iron (Fe) content of rice plants. To investigate whether planthopper attacks change rice Fe homeostasis at the molecular level, the response of rice Fe homeostasis to early feeding by small brown planthopper (SBPH) was examined by transcriptome profiling. Results showed that the concentration of Fe and nicotianamine decreased in resistant rice genotype and increased in susceptible rice genotype after attack by SBPH. Transcriptome profiling showed that 13 and 21 Fe homeostasis-related genes encoded enzymes that were involved in phytosiderophore biosynthesis and that Fe transporters and regulators displayed altered expression in resistant and susceptible rice genotypes, respectively, after attack by SBPH. This revealing response of Fe homeostasis to planthopper attack in rice at the molecular level provided new insight into rice plants' response to planthopper attack and uncovered a novel physiological response of rice to planthopper attack, which extended our knowledge of the early interaction between rice and SBPH.

Phytosiderophores revisited: 2′-deoxymugineic acid-mediated iron uptake triggers nitrogen assimilation in rice (Oryza sativa L.) seedlings

Poaceae plants release phytosiderophores into the rhizosphere in order to chelate iron (Fe), which often exists in insoluble forms especially under high pH conditions. The impact of phytosiderophore treatment at the physiological and molecular levels in vivo remains largely elusive, although the biosynthesis of phytosiderophores and the transport of phytosiderophore-metal complexes have been well studied. We recently showed that the application of 30 μM of the chemically synthesized phytosiderophore 2′-deoxymugineic acid (DMA) was sufficient for apparent full recovery of otherwise considerably reduced growth of hydroponic rice seedlings at high pH. Moreover, unexpected induction of high-affinity nitrate transporter gene expression as well as nitrate reductase activity indicates that the nitrate response is linked to Fe homeostasis. These data shed light on the biological relevance of DMA not simply as a Fe chelator, but also as a trigger that promotes plant growth by reinforcing nitrate assimilation.

Chlorophyll biosynthesis as the basis of iron use efficiency under iron deficiency and its relationship with the phytosiderophore synthesis and release in wheat

The present study demonstrates that chlorophyll biosynthesis can be effectively used for identifying iron (Fe) deficiency tolerant cultivars from a large population. Fifty genetically diverse wheat cultivars were screened on the basis of the leaf greenness index, i.e. chlorophyll retention under iron deficient (Fe−) compared to iron sufficient (Fe+) conditions, and three Fe deficiency tolerant and two Fe deficiency susceptible cultivars were identified. These two groups of cultivars were further tested for their efficiency for Fe uptake and translocation and biomass production per unit Fe under Fe deficient condition in the nutrient solution culture. Fe deficiency tolerant lines were found capable of translocating more Fe to the shoot and produced greater biomass per unit of Fe when compared with the susceptible group. Efficiency for chlorophyll biosynthesis per unit shoot Fe was also higher for the tolerant group of cultivars. Fe use efficient cultivars produced and released a larger amount of phytosiderophore (PS) and differed from the inefficient cultivars in terms of the PS release but not in the PS biosynthesis. Thus, indicating that the limitation at the level of release of the PS was 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 genotypic variation in the Fe use efficiency.

Comparative Proteomics Analysis of the Rice Roots Colonized byHerbaspirillum seropedicaeStrain SmR1 Reveals Induction of the Methionine Recycling in the Plant Host

Although the use of plant growth-promoting bacteria in agriculture is a reality, the molecular basis of plant-bacterial interaction is still poorly understood. We used a proteomic approach to study the mechanisms of interaction of Herbaspirillum seropedicae SmR1 with rice. Root proteins of rice seedlings inoculated or noninoculated with H. seropedicae were separated by 2-D electrophoresis. Differentially expressed proteins were identified by MALDI-TOF/TOF and MASCOT program. Among the identified proteins of H. seropedicae, the dinitrogenase reductase NifH and glutamine synthetase GlnA, which participate in nitrogen fixation and ammonium assimilation, respectively, were the most abundant. The rice proteins up-regulated included the S-adenosylmethionine synthetase, methylthioribose kinase, and acireductone dioxygenase 1, all of which are involved in the methionine recycling. S-Adenosylmethionine synthetase catalyzes the synthesis of S-adenosylmethionine, an intermediate used in transmethylation reactions and in ethylene, polyamine, and phytosiderophore biosynthesis. RT-qPCR analysis also confirmed that the methionine recycling and phytosiderophore biosynthesis genes were up-regulated, while ACC oxidase mRNA level was down-regulated in rice roots colonized by bacteria. In agreement with these results, ethylene production was reduced approximately three-fold in rice roots colonized by H. seropedicae. The results suggest that H. seropedicae stimulates methionine recycling and phytosiderophore synthesis and diminishes ethylene synthesis in rice roots.

Rice genes involved in phytosiderophore biosynthesis are synchronously regulated during the early stages of iron deficiency in roots

BackgroundThe rice transcription factors IDEF1, IDEF2, and OsIRO2 have been identified as key regulators of the genes that control iron (Fe) uptake, including the biosynthesis of mugineic acid-family phytosiderophores (MAs). To clarify the onset of Fe deficiency, changes in gene expression were examined by microarray analysis using rice roots at 3, 6, 9, 12, 24, and 36 h after the onset of Fe-deficiency treatment.ResultsMore than 1000 genes were found to be upregulated over a time course of 36 h. Expression of MAs-biosynthetic genes, OsIRO2, and the Fe3+–MAs complex transporter OsYSL15 was upregulated at the 24 h and 36 h time points. Moreover, these genes showed very similar patterns of expression changes, but their expression patterns were completely different from those of a metallothionein gene (OsIDS1) and the Fe2+-transporter genes OsIRT1 and OsIRT2. OsIDS1 expression was upregulated by the 6 h time point. The early induction of OsIDS1 expression was distinct from the other Fe-deficiency-inducible genes investigated and suggested a functional relationship with heavy-metal homeostasis during the early stages of Fe deficiency.ConclusionsWe showed that many genes related to MAs biosynthesis and transports were regulated by a distinct mechanism in roots. Furthermore, differences in expression changes and timing in response to Fe deficiency implied that different combinations of gene regulation mechanisms control the initial responses to Fe deficiency.Electronic supplementary materialThe online version of this article (doi:10.1186/1939-8433-6-16) contains supplementary material, which is available to authorized users.

Characterizing the Crucial Components of Iron Homeostasis in the Maize Mutants ys1 and ys3

To acquire iron (Fe), graminaceous plants secrete mugineic acid family phytosiderophores through the phytosiderophore efflux transporter TOM1 and take up Fe in the form of Fe(III)–phytosiderophore complexes. Yellow stripe 1 (ys1) and ys3 are recessive mutants of maize (Zea mays L.) that show typical symptoms of Fe deficiency, i.e., interveinal chlorosis of the leaves. The ys1 mutant is defective in the Fe(III)–phytosiderophore transporter YS1 and is therefore unable to take up Fe(III)–phytosiderophore complexes. While the ys3 mutant has been shown to be defective in phytosiderophores release, the causative gene has not been identified. The present study was performed to characterize the expression profiles of the genes in ys1 and ys3 mutants to extend our understanding of Fe homeostasis in maize. Using quantitative real-time polymerase chain reaction, we assessed changes in the levels of gene expression in response to Fe deficiency of genes involved in Fe homeostasis, such as those related to phytosiderophore biosynthesis and Fe transport. As with other crops, these Fe deficiency-inducible genes were also upregulated in maize. In addition, these Fe deficiency-inducible genes were upregulated in both the ys1 and ys3 mutants, even under Fe-sufficient conditions. Indeed, the Fe concentrations in the roots of ys1 and ys3 plants were lower than that of wild-type controls. These results suggest that ys1 and ys3 are Fe-deficient during growth in the presence of Fe. In agreement with previous reports, the level of YS1 expression decreased in the ys1 mutant. Moreover, the expression level of a homolog of TOM1 in maize decreased significantly in the ys3 mutant. Unspliced introns of ZmTOM1 were detected only in ys3, and not in YS1YS3 or ys1, suggesting that ZmTOM1 may be involved in the ys3 phenotype.

Senescence‐induced iron mobilization in source leaves of barley (Hordeum vulgare) plants

• Retranslocation of iron (Fe) from source leaves to sinks requires soluble Fe binding forms. As much of the Fe is protein-bound and associated with the leaf nitrogen (N) status, we investigated the role of N in Fe mobilization and retranslocation under N deficiency- vs dark-induced leaf senescence. • By excluding Fe retranslocation from the apoplastic root pool, Fe concentrations in source and sink leaves from hydroponically grown barley (Hordeum vulgare) plants were determined in parallel with the concentrations of potential Fe chelators and the expression of genes involved in phytosiderophore biosynthesis. • N supply showed opposing effects on Fe pools in source leaves, inhibiting Fe export out of source leaves under N sufficiency but stimulating Fe export from source leaves under N deficiency, which partially alleviated Fe deficiency-induced chlorosis. Both triggers of leaf senescence, shading and N deficiency, enhanced NICOTIANAMINE SYNTHASE2 gene expression, soluble Fe pools in source leaves, and phytosiderophore and citrate rather than nicotianamine concentrations. • These results indicate that Fe mobilization within senescing leaves is independent of a concomitant N sink in young leaves and that phytosiderophores enhance Fe solubility in senescing source leaves, favoring subsequent Fe retranslocation.

Response of barley plants to Fe deficiency and Cd contamination as affected by S starvation

Both Fe deficiency and Cd exposure induce rapid changes in the S nutritional requirement of plants. The aim of this work was to characterize the strategies adopted by plants to cope with both Fe deficiency (release of phytosiderophores) and Cd contamination [production of glutathione (GSH) and phytochelatins] when grown under conditions of limited S supply. Experiments were performed in hydroponics, using barley plants grown under S sufficiency (1.2 mM sulphate) and S deficiency (0 mM sulphate), with or without Fe(III)-EDTA at 0.08 mM for 11 d and subsequently exposed to 0.05 mM Cd for 24 h or 72 h. In S-sufficient plants, Fe deficiency enhanced both root and shoot Cd concentrations and increased GSH and phytochelatin levels. In S-deficient plants, Fe starvation caused a slight increase in Cd concentration, but this change was accompanied neither by an increase in GSH nor by an accumulation of phytochelatins. Release of phytosiderophores, only detectable in Fe-deficient plants, was strongly decreased by S deficiency and further reduced after Cd treatment. In roots Cd exposure increased the expression of the high affinity sulphate transporter gene (HvST1) regardless of the S supply, and the expression of the Fe deficiency-responsive genes, HvYS1 and HvIDS2, irrespective of Fe supply. In conclusion, adequate S availability is necessary to cope with Fe deficiency and Cd toxicity in barley plants. Moreover, it appears that in Fe-deficient plants grown in the presence of Cd with limited S supply, sulphur may be preferentially employed in the pathway for biosynthesis of phytosiderophores, rather than for phytochelatin production.

Bio‐available zinc in rice seeds is increased by activation tagging of nicotianamine synthase

We generated rice lines with increased content of nicotianamine (NA), a key ligand for metal transport and homeostasis. This was accomplished by activation tagging of rice nicotianamine synthase 2 (OsNAS2). Enhanced expression of the gene resulted in elevated NA levels, greater Zn accumulations and improved plant tolerance to a Zn deficiency. Expression of Zn-uptake genes and those for the biosynthesis of phytosiderophores (PS) were increased in transgenic plants. This suggests that the higher amount of NA led to greater exudation of PS from the roots, as well as stimulated Zn uptake, translocation and seed-loading. In the endosperm, the OsNAS2 activation-tagged line contained up to 20-fold more NA and 2.7-fold more zinc. Liquid chromatography combined with inductively coupled plasma mass spectrometry revealed that the total content of zinc complexed with NA and 2'-deoxymugineic acid was increased 16-fold. Mice fed with OsNAS2-D1 seeds recovered more rapidly from a zinc deficiency than did control mice receiving WT seeds. These results demonstrate that the level of bio-available zinc in rice grains can be enhanced significantly by activation tagging of OsNAS2.

Increased sensitivity to iron deficiency in Arabidopsis thaliana overaccumulating nicotianamine

Nicotianamine (NA) is a non-protein amino acid derivative synthesized from S-adenosyl L-methionine able to bind several metal ions such as iron, copper, manganese, zinc, or nickel. In plants, NA appears to be involved in iron availability and is essential for the plant to complete its biological cycle. In graminaceous plants, NA is also the precursor in the biosynthesis of phytosiderophores. Arabidopsis lines accumulating 4- and 100-fold more NA than wild-type plants were used in order to evaluate the impact of such an NA overaccumulation on iron homeostasis. The expression of iron-regulated genes including the IRT1/FRO2 iron uptake system is highly induced at the transcript level under both iron-sufficient and iron-deficient conditions. Nevertheless, NA overaccumulation does not interfere with the iron uptake mechanisms since the iron levels are similar in the NA-overaccumulating line and wild-type plants in both roots and leaves under both sufficient and deficient conditions. This observation also suggests that the translocation of iron from the root to the shoot is not affected in the NA-overaccumulating line. However, NA overaccumulation triggers an enhanced sensitivity to iron starvation, associated with a decrease in iron availability. This study draws attention to a particular phenotype where NA in excess paradoxically leads to iron deficiency, probably because of an increase of the NA apoplastic pool sequestering iron. This finding strengthens the notion that extracellular NA in the apoplast could be a major checkpoint to control plant iron homeostasis.

Promoter analysis of iron-deficiency-inducible barley IDS3 gene in Arabidopsis and tobacco plants

Under conditions of iron deficiency, graminaceous plants induce the expression of genes involved in the biosynthesis of mugineic acid family phytosiderophores. We previously identified the novel cis-acting elements IDE1 and IDE2 (iron-deficiency-responsive element 1 and 2) through promoter analysis of the barley ( Hordeum vulgare L.) iron-deficiency-inducible IDS2 gene in tobacco ( Nicotiana tabacum L.). To gain further insight into plant gene regulation under iron deficiency, we analyzed the barley iron-deficiency-inducible IDS3 gene, which encodes mugineic acid synthase. IDS3 promoter fragments were fused to the β-glucuronidase ( GUS) gene, and this construct was introduced into Arabidopsis thaliana L. and tobacco plants. In both Arabidopsis and tobacco, GUS activity driven by the IDS3 promoter showed strongly iron-deficiency-inducible and root-specific expression. Expression occurred mainly in the epidermis of Arabidopsis roots, whereas expression was dominant in the pericycle, endodermis, and cortex of tobacco roots, resembling the expression pattern conferred by IDE1 and IDE2. Deletion analysis revealed that a sequence within −305 nucleotides from the translation start site was sufficient for specific expression in both Arabidopsis and tobacco roots. Gain-of-function analysis revealed functional regions at −305/−169 and −168/−93, whose coexistence was required for the induction activity in Arabidopsis roots. Multiple IDE-like sequences were distributed in the IDS3 promoter and were especially abundant within the functional region at −305/−169. A sequence moderately homologous to that of IDE1 was also present within the −168/−93 region. These IDE-like sequences would be the first candidates for the functional iron-deficiency-responsive elements in the IDS3 promoter.

OsMTN encodes a 5′-methylthioadenosine nucleosidase that is up-regulated during submergence-induced ethylene synthesis in rice (Oryza sativa L.)

Methylthioadenosine (MTA) is released as a by-product of S-adenosylmethionine (AdoMet)-dependent reactions central to ethylene, polyamine, or phytosiderophore biosynthesis. MTA is hydrolysed by methylthioadenosine nucleosidase (MTN; EC 3.2.2.16) into adenine and methylthioribose which is processed through the methionine (Met) cycle to produce a new molecule of AdoMet. In deepwater rice, submergence enhances ethylene biosynthesis, and ethylene in turn influences the methionine cycle through positive feedback regulation of the acireductone dioxygenase gene OsARD1. In rice, MTN is encoded by a single gene designated OsMTN. Recombinant OsMTN enzyme had a KM for MTA of 2.1 mM and accepted a wide array of 5' substitutions of the substrate. OsMTN also metabolized S-adenosylhomocysteine (AdoHcy) with 15.9% the rate of MTA. OsMTN transcripts and OsMTN-specific activity increased slowly and in parallel upon submergence, indicating that regulation occurred mainly at the transcriptional level. Neither ethylene, MTA, nor Met regulated OsMTN expression. Analysis of steady-state metabolite levels showed that MTN activity was sufficiently high to prevent Met and AdoMet depletion during long-term ethylene biosynthesis.

Iron deficiency induces sulfate uptake and modulates redistribution of reduced sulfur pool in barley plants.

We studied the possibility that the sulfur (S) assimilatory pathway might be modulated by iron (Fe) starvation in barley, as a consequence of plant requirement for an adequate amount of reduced S to maintain methionine and, in turn, phytosiderophore biosynthesis. Barley seedlings were grown with or without 100 µm FeIII-EDTA, at three S levels in the nutrient solution (S2 = 1200, S1 = 60, and S0 = 0 µm sulfate) in order to reproduce conditions of optimal supply, latent and severe deficiency, respectively. Fe deprivation increased root cysteine content irrespective of the S supply. However, this increase was not associated with either higher rates of 35SO42- uptake or increased expression of the gene for the high-affinity sulfate transporter, HvST1, and these roots failed to increase their activities of ATP sulfurylase (ATPS) and O-acetylserine(thiol) lyase (OASTL). We observed a significant increase in 35SO42- uptake rate (+76%) only in Fe-deficient S1 plants and we found an increase in root ATPS activity only in S0 plants. We observed an increase of ATPS enzyme activity in leaves of S1 and S2 plants, most likely suggesting increased S assimilation followed by translocation of thiols (Cys) to the root. Taken together, our results suggest that Fe deficiency affects the partitioning from the shoot to the root of the reduced S pool within the plant and can affect SO42- uptake under limited S supply.

Real-time [11C]methionine translocation in barley in relation to mugineic acid phytosiderophore biosynthesis.

[11C]Methionine was supplied through barley roots and the 11C signal was followed for 90 min using a real-time imaging system (PETIS), with subsequent development of autoradiographic images of the whole plant. In all cases, [11C]methionine was first translocated to the 'discrimination center', the basal part of the shoot, and this part was most strongly labeled. Methionine absorbed by the roots of the plants was subsequently translocated to other parts of the plant. In Fe-deficient barley plants, a drastic reduction in [11C]methionine translocation from the roots to the shoot was observed, while a greater amount of 11C was found in the leaves of Fe-sufficient or methionine-pretreated Fe-deficient plants. Treatment of Fe-deficient plants with aminooxyacetic acid, an inhibitor of nicotianamine aminotransferase, increased the translocation of [11C]methionine to the shoot. The retention of exogenously supplied [11C]methionine in the roots of Fe-deficient barley indicates that the methionine is used in the biosynthesis of mugineic acid phytosiderophores in barley roots. This and the absence of methionine movement from shoots to the roots suggest that the mugineic acid precursor methionine originates in the roots of plants.

Enhanced tolerance of rice to low iron availability in alkaline soils using barley nicotianamine aminotransferase genes.

One of the widest ranging abiotic stresses in world agriculture arises from low iron (Fe) availability due to high soil pH, with 30% of arable land too alkaline for optimal crop production. Rice is especially susceptible to low iron supply, whereas other graminaceous crops such as barley are not. A barley genomic DNA fragment containing two naat genes, which encode crucial enzymes involved in the biosynthesis of phytosiderophores, was introduced into rice using Agrobacterium-mediated transformation and pBIGRZ1. Phytosiderophores are natural iron chelators that graminaceous plants secrete from their roots to solubilize iron in the soil. The two transgenes were expressed in response to low iron nutritional status in both the shoots and roots of rice transformants. Transgenic rice expressing the two genes showed a higher nicotianamine aminotransferase activity and secreted larger amounts of phytosiderophores than nontransformants under iron-deficient conditions. Consequently, the transgenic rice showed an enhanced tolerance to low iron availability and had 4.1 times greater grain yields than that of the nontransformant rice in an alkaline soil.

Mining out for iron

Although iron is an abundant element in most soils, its availability for plants can be limiting, especially in soils with a basic pH. From an agricultural point of view, iron deficiency is responsible for severe losses owing to chlorosis, decreasing the yield and nutritional value of the crops. Plants have evolved two main strategies to cope with low iron availability. The first one, used by most plants, but not by grasses, relies on the reduction of soluble iron(III) chelates and subsequent uptake of iron(II) by root transporters. By contrast, grasses have evolved a mechanism based on the release of small molecules into the rhizosphere that efficiently chelates iron(III), the phytosiderophores and subsequent iron import by uptake of the iron(III)-phytosiderophore complexes into root cells.If we want to understand and improve iron nutrition in major grass crops, such as rice, sorghum, wheat and corn, it is crucial to identify the components involved in iron uptake mechanisms at the molecular level.In the past few years, the genes encoding the key enzymes involved in phytosiderophore biosynthesis have been characterized. Catherine Curie and colleagues1xMaize yellow stripe 1 encodes a protein directly involved in Fe(III) uptake. Curie, C et al. Nature. 2001; 409: 346–349Crossref | PubMed | Scopus (441)See all References1 report the identification of a new gene family encoding the transporters responsible for the uptake of iron siderophore complexes from the rhizosphere to the root cells. Earlier physiological studies have shown that iron-siderophore complex uptake of the iron-deficient maize mutant, yellow stripe 1 (ys1), is impaired. The authors have taken advantage of transposon-tagged alleles of ys1 to identify the disrupted gene. They found that YS1 encodes a protein with 12 transmembrane domains. Furthermore, expression of this protein in a yeast mutant defective for iron transport, restores growth of this yeast strain on medium with low levels of iron, but only in the presence of phytosiderophores. Thus, YS1 encodes a maize iron-siderophore complex uptake transporter. The discovery of this gene family should help the selection or engineering of iron-efficient crop species. Interestingly, the occurrence of YS1-like genes in non-grass species suggests that this type of transporter could play a more general role in iron homeostasis in plants.