Chem. Rev.

2009, 109, 4553–4567 4553

Iron Uptake and Transport in Plants: The Good, the Bad, and the Ionome

Joe Morrissey and Mary Lou Guerinot*
Department of Biological Sciences, Dartmouth College, Hanover, New Hampshire 03755

Received March 25, 2009

Contents 6. Conclusions and Future Directions 4564
7. Acknowledgments 4565
1. Introduction 4553 8. References 4565
2. Fe Uptake 4553
2.1. Reduction-Based Strategy 4554 1. Introduction
2.1.1. Toxic Metals and Fe Deficiency 4554
2.1.2. Sequestration and Buffering of Metal Influx 4555 Fe is essential for plant growth. At the same time, Fe is
2.1.3. Uptake of Apoplastic Fe 4556 highly reactive and toxic via the Fenton reaction. Conse-
quently, plants tightly control Fe homeostasis and react to
2.2. Chelation-Based Strategy 4556
Fe deficiency as well as Fe overload. The ability of plants
2.2.1. Yellow-Stripe 1 4556 to respond to Fe availability ultimately affects human
2.2.2. Chelation and Toxic Metals 4556 nutrition, both in terms of crop yield and the Fe concentration
2.3. Combination of Reduction and Chelation 4557 of edible tissues. Thus, elucidating the mechanisms of Fe
Strategies uptake and transport is essential for the breeding of crops
3. Long-Distance Fe Transport 4557 that are more nutrient rich and more tolerant of Fe-limited
3.1. Xylem 4558 soils.
3.2. Phloem 4558 This review covers Fe transport and homeostasis in plants,
3.2.1. NA and YSLs 4558 focusing on the research published in the past five years.
3.2.2. Nicotianamine 4558 Because Fe transporters often have a broad range of
3.2.3. NA Levels and Fe Localization 4559 substrates, we also examine the relationship between Fe and
3.2.4. NA and Ni Tolerance 4559 the toxic metals that often accompany Fe uptake, namely,
3.2.5. YSLs and Long-Distance Fe Transport in 4559 Cd, Co, and Ni. We begin by discussing Fe uptake into the
Arabidopsis root, then long-distance transport to the shoot, and finally,
3.2.6. OsIRT1 4559 the loading of Fe into seeds. And, because Fe is essential to
3.2.7. ITP 4559 the metabolism of the mitochondria and chloroplast, we also
3.3. Control of Long-Distance Fe Transport in 4560 look at the recent discoveries in Fe transport and homeostasis
Barley at the intracellular level. We do not cover the regulation of
4. Fe and Seeds 4560 these transporters because this topic has been recently
4.1. Loading of Fe 4560 reviewed.1
4.1.1. NA, YSL1, and YSL3 4560
4.1.2. OPT3 4561 2. Fe Uptake
4.2. Storage of Fe 4561 Plants mainly acquire Fe from the rhizosphere. Although
4.2.1. VIT1 4561 Fe is one of the most abundant metals in the earth’s crust,
4.2.2. NRAMP3 and NRAMP4 4561 its availability to plant roots is very low. Fe availability is
4.2.3. FER2 4562 dictated by the soil redox potential and pH. In soils that are
4.3. Fe Bioavailability for Humans 4562 aerobic or of higher pH, Fe is readily oxidized, and is
5. Intracellular Fe 4563 predominately in the form of insoluble ferric oxides. At lower
pH, the ferric Fe is freed from the oxide and becomes more
5.1. Plastids 4563
available for uptake by roots. Because 30% of the world’s
5.1.1. FRO7 4563 cropland is too alkaline for optimal plant growth,2 and some
5.1.2. PIC1 4563 staple crops, like rice, are especially susceptible to Fe
5.1.3. Ferritin 4563 deficiency,3 much research has focused on how plants cope
5.2. Mitochondria 4563 with Fe limitation.
5.2.1. Ferritin and Frataxin 4563 The responses to Fe deficiency include changes in root
5.2.2. ATM3 4564 morphology2 and up-regulation of genes involved in Fe
5.3. Vacuole 4564 uptake.4,5 In fact, in Arabidopsis thaliana, up to 85% of the
5.3.1. NA and the Vacuole 4564 genes expressed in particular regions of the root are differ-
5.3.2. VIT1, NRAMP3, and NRAMP4 4564 entially regulated by Fe.4 This transcriptome analysis was
made possible by the isolation, via fluorescence activated
* To whom correspondence should be addressed. E-mail: mary.lou.guerinot@ cell sorting analysis, of cells from specific root layers that
dartmouth.edu. Phone: 603-646-2527. were expressing GFP under the control of cell-specific
10.1021/cr900112r CCC: $71.50  2009 American Chemical Society
Published on Web 09/16/2009

uptake of Fe.4554 Chemical Reviews. protons are released into the rhizosphere. the transgenic overexpression of ferric of the Faculty for the Sciences from 1998 to 2001 and as Vice Provost chelate reductases in the roots of rice.11. a 5000-member organization Additional root epidermal transporters for these metals have devoted to the advancement of plant science. Mn. Mn. Zn. while grasses secrete phytosiderophores (PSs) that readily bind Fe3+. While AHA1. and mediate the appropriate Joe Morrissey received his undergraduate education at the University of Minnesota and performed research in John Ward’s laboratory.7-10 but it is best charac- terized in Arabidopsis (Figure 2). and the Fe-PS complexes are then transported back into the roots. Of the the biology Ph. 109.D.23 and Ni.1. accessible for uptake by a Fe2+ transporter. She has served on the not yet been identified. Guerinot is a molecular geneticist whose principal has been successful in increasing tolerance to Fe-limiting expertise and research interests are in the area of metal transport and conditions.13 The expression level of AHA2 is highest among the three. This allows detection of differential expression profiles among specific cell types that 2. which complement yeast mutants defective in the graduate students.6 The transcript levels within each layer were then all of which are potentially toxic. Mn. and serves on the Board of Directors for TAIR (The Arabidopsis Information Resource).5 AHA2 is the primary root H+-ATPase in the Fe deficiency response. served as the associate dean Arabidopsis. in biology from Dalhousie ciency.14 professor in 1985. but in the irt1-1 loss of function Advisory Committee for the Biological Sciences Directorate at NSF. while genes related to root hair morphogenesis were down-regulated. She was chair of the Department This appears to be the rate-limiting step in Fe uptake in of Biological Sciences from 1994 to 1998.7 Thus. is a mutant. it was found that the vast majority of the transcriptome is altered by environmental stress and that these changes are most dramatic in the root epidermis. She teaches Microbiology found to be transported by the tomato orthologs LeIRT1 and and Molecular Genetics and has mentored numerous undergraduates. she has been systematically dissecting how iron gets from the “soil to the seed” and Once reduced. Zn. In particular. two specific strategies of Fe uptake lab of Mary Lou Guerinot. program at Dartmouth College in 2004 and joined the changes in the epidermis. making Fe more soluble. His graduate studies are focused on have been identified in plants. Electrons are transferred from the Department of Biological Sciences at Dartmouth as an assistant NADH+ across four heme groups to Fe in the rhizosphere. After completing postdoctoral studies at the University of The NADPH-dependent ferric chelate reductase. this core may define the essential features of each cell type.20. tobacco. the Fe deficiency response also leads to the uptake of metals other than Fe. Nongraminaceous plants characterizing the function of the ferroportin genes in Arabidopsis and reduce Fe3+ via a membrane-bound reductase to make it using ionomic screens to identify genes involved in metal homeostasis.16-18 regulation of gene expression by metals.4. promoters.24 of the American Society of Plant Biologists. She earned her bachelor’s degree in found to reduce rhizosphere acidification during Fe defi- biology at Cornell University and her Ph.1. while regulation of Fe levels in the root is facilitated by modulating uptake in the epidermis. 2009. and soybeans from 2001 to 2004.13 University. The expression of genes related to metal transport and chelation was increased in the epidermis. Interestingly. A similarly broad range of metals was of Applied and Environmental Microbiology. Guerinot is a fellow of the American epidermal cells by the divalent metal transporter AtIRT1. and only the loss of AHA2 was Mary Lou Guerinot is the Ronald and Deborah Harris Professor in the Sciences at Dartmouth College. and Co decreases member of the Scientific Advisory Board for the Donald Danforth Plant significantly.19-21 Association for the Advancement of Science and has served as President AtIRT1 also transports Zn. She was promoted to an associate professor with tenure in 1991 and to full professor in 1997. indicating layer-specific roles Fe deficiency.12 This lowers the soil pH.1. AHA2. She is currently an associate editor gesting that AtIRT1 is a primary transporter for these metals of Plant Cell and Environment as well as a member of the editorial board under Fe deficiency. Cd. Toxic Metals and Fe Deficiency cannot be seen when the root as a whole is examined. These results suggest that sensing of Fe levels and control of the Fe deficiency response occurs in the vasculature. No.25 sug- Science Center. In response to Fe defi- ciency. Fe(II) can then be transported into the root has identified many key genes. by AHA H+-ATPases expressed in the epidermis. in the stele. and the plants become Cd tolerant. Cd interferes with Fe movement from root to . and AHA7 are all up-regulated in the root epidermis in response to Fe deficiency. AtFRO2. When these Fe deficiency-induced changes were compared with the response to salt stress. Vol. measured via microarray analysis. there is also a small set of genes unaffected by either stress. Reduction-Based Strategy Components of the reduction strategy have been described in many nongraminaceous species.15 In fact. and Cu.D. He entered transcriptional responses to environmental stresses.22. Large transcriptional differences between layers in response to Fe The presence of Cd and Co have been shown to exacerbate deficiency were identified. 2. and postdoctoral researchers. 10 Morrissey and Guerinot (Figure 1). genes associated with signaling and stress responses were up-regulated. LeIRT2. Maryland and at the DOE-MSU Plant Research Laboratory. shoot accumulation of Fe. Co. she joined then reduces Fe3+ to Fe2+.

30 Mutations that destroyed IRT1 function were almost exclusively found in the two R-helices that compose the fourth and fifth transmembrane domains. by Fe toxicity. Within the epidermis. localizes to the vacuolar mem- Cd increased. rather than uptake.32 Inhibition of root elongation in the Arabi- The uptake of Cd during Fe deficiency is of special dopsis ecotype Col-0 was known to occur during phosphorus interest. and is thought to sequester Zn in the vacuole during Fe access to the phloem. could modulate selec- tivity of Fe. 2009. Because ingestion of plants was likely no longer complexed with phosphate. because Cd is considered one of the most toxic crop limitation and was believed to be caused by a phosphorus- contaminants.Iron Uptake and Transport in Plants Chemical Reviews. the soil by simply removing Fe from the growth medium. attributed to phosphorus deficiency was found to be caused treated plants (Morrissey and Guerinot. It is treated with Co: Fe uptake increased. Fe deficiency. 2008 The American Association for the Advancement of Science (ref 4). The ferric The influx of toxic metals via IRT1 is counteracted in part chelate reductase FRO2 is expressed. producing variants optimized to biofortify crops while excluding toxic contaminants. Cor ) cortex. Sequestration and Buffering of Metal Influx Fe deficiency in nongraminaceous species. In response to 2. greatly is the primary route of Cd exposure for nonsmokers. A similar role has been described for MTP3. especially the first extracellular loop. 10 4555 Figure 1. Ulti- mately. pattern and root localization of FPN2 suggests that it serves as an adaptation to the influx of Ni and Co during Fe shoot.29 efforts increasing its bioavailability.28 Thus. phloem saps also decreased significantly. Expression of mutagenized IRT1 in yeast found that FPN2. unpub- become Fe starved. FPN2 is localized to the vacuolar membrane24 metal transporter IRT1. copyright 2001 Nature Publishing Group (ref 153). A similar phenotype was seen in mung bean seedlings Fe itself is highly reactive and potentially toxic. Ste ) stele. The influx of Fe via IRT1 may have been made to understand and modify the selectivity of be buffered in the outer root layers by the expression of IRT1. protons are exuded into the rhizosphere. QC ) quiescent center. unpublished data). Fe uptake from soil. This suggests that the presence of Cd impairs brane.2. Vol. it is believed that this region forms a metal binding pocket that facilitates ion movement across the membrane. a phenotype long deficiency response in Arabidopsis is up-regulated in Co. Zn. End ) endodermis. Mn.1. RNase. this demonstrates that the broad substrate range of Fe transporters can be adjusted. Fe presumably moves out of the epidermis via the Guerinot. but buffering move from the root to the shoot. presumably .153 (B) Enriched Gene-Ontology categories: miRNA.31 napus. Cd treatment of Brassica napus produces a dramatic deficiency. Transcriptional changes in response to Fe deficiency in specific root layers. Cei ) cortex/endodermis initial. Figure 2.26 The level of Fe in the xylem and lished data). while the level of which is also Fe-regulated. Indeed. reducing Fe(III) to Fe(II). GTPase. microRNA. No.4 Reprinted with permission. most likely by the AHA2 H+-ATPase. reduction strategy. (A) Root layers marked by propidium iodide staining of the cell wall (red) and expression of GFP in the stele and endodermis. accordingly. the Fe Fe uptake is clearly important. Epi ) epidermis. the divalent metal in the two outermost root layers of Arabidopsis and appears effluxer FPN2 is expressed during Fe deficiency on the vacuolar membrane and may serve to buffer Fe uptake by sequestering excess to sequester metals in the vacuole. guanosine triphosphatase. ribonuclease. the loss of FPN2 results in increased increase in Fe accumulation in the roots. at least in B. Cd has the opposite bioavailability profile sensing regulatory pathway. The conditions that trigger IRT1 expression also enhance Cd inhibition was actually caused by the toxic effects of Fe that availability for uptake by IRT1. The Fe-regulated expression plasmodesmata.27 Additionally. by the expression of the metal effluxer FPN2 (IREG2) during which can then be transported into the root epidermis by the divalent Fe deficiency. but Fe was unable to unclear which ligands bind Fe after transport. amino acid substitutions in the first third of the protein. it was recently shown compared with Fe: in aerobic conditions where Fe is oxidized that root growth is restored during phosphorus deficiency and insoluble. and Cd. The Fe may be bound by phytate or NA in FPN2 confers tolerance to Ni24 and Co (Morrissey and the vacuole. When expressed in yeast. while the shoots sensitivity to Ni and Co (Morrissey and Guerinot. unpublished data). Instead. free Fe in the vacuole. which transports Fe (unpublished data). Cd becomes more soluble. 109.

expression of the between the sixth and seventh transmembrane regions OsIRO2 transcription factor increases dramatically over the provided the selectivity. Ferric chelate reductase activity and proton extrusion also ZmYS1 is expressed in roots in response to Fe deficiency. To species. the reduction strategy. in nongraminaceous species. deficiency response in maize and results in reduced Fe levels For instance.43 uptake.4556 Chemical Reviews. In response to This pool decreases when a plant becomes Fe deficient. phenolics were filtered from the liquid japonica. 2009. to regreen. and while into the root epidermis via a high-affinity uptake system.34 2. the genes required for sulfur uptake. AtYSL3. The resulting Fe(III)-PS complexes are readily transported Maize DMA also appears to bind Cd in soil. Uptake of Apoplastic Fe Another aspect of the Fe deficiency response in non- graminaceous plants is the secretion of phenolic compounds into the rhizosphere2 and the uptake of apoplastic Fe. Whether an Fe-phenolic complex is directly is Fe limiting for plants. Under normal growth conditions. A candidate would be the that transport of Fe-PS is still efficient at very high pH. 10 Morrissey and Guerinot sequestering excess Fe in the vacuole. barley. and siderophores with a high affinity for Fe3+. and although initially chlorotic. and PS synthesis are dramatically up-regulated in tween the two transporters showed that the extracellular loop the first 24 h of Fe deficiency. No. Vol.1. the HvYS1 peptide formed an R helix in solution.35 because the negatively charged carboxyl groups of the cell walls serve as a cation sink.47 However. methionine ZmYS1. suggesting mobilization into the symplast.2. the expression of ZmYS1 in oocytes showed root epidermis of nongrasses.39 which are adapted for growing in anaerobic soils determine whether this is an essential component of the Fe where Fe is more soluble.2. and Ni at the same Grasses depend on the uptake of Fe chelated by soluble rate as Fe-PS. which transports Fe and Zn33 and has been localized to vesicles in the epidermis. The maize PS deoxymugineic acid (DMA) also readily 2.46 When the loops were synthesized course of the first 5 days of Fe starvation and is believed to in Vitro. the type of environment that symplast. which is up- regulated in the root epidermis under Fe deficiency. in the xylem sap. which is 95% similar to barley. This produced plants that were much The most well-characterized Fe-PS transporter is the more chlorotic and could not recover from Fe deficiency. only transports Fe-PS. allowing transport into the root adapted to high pH solutions. because potential Fe more difficult with increasing soil pH due to the pH optimum chelate transporters have not yet been characterized in the of the reductase. and there is a strong correlation between presence of Cd in growth media up-regulates the Fe the volume of PS released and resistance to Fe limiting soils. activate the expression of genes related to PS synthesis and while the ZmYS1 peptide remained flexible.36 How this Fe is The PS readily chelate Fe3+. deficiency response. like IRT1 Fe deficiency. and OsYSL15 in rice). including those toxic to plants (Figure 3). It has been observed that as much as 75% of Fe in the roots is attached to the apoplast. although it is unclear how phenolics facilitate in a yellowing between the veins (interveinal chlorosis). 109. suggesting that Fe uptake.2 Figure 3. and the recovery from Fe deficiency. PSs are synthesized and secreted into the rhizosphere. the leaves began soil. HvYS1. Fe(II). Cu.2.42.2.38 In and humans. the growth medium by constant recirculation through a resin overexpression of enzymes in the barley PS synthesis column. The filtering of phenolics. Chelation-Based Strategy chelates other metals. suggesting .45 Domain swapping be- synthesis. Perhaps phenolics mediate extraction of Fe from the The transport of Fe-PS by ZmYS1 appears to be well- negatively charged cell walls.2 In response to Fe(III) complexed with the PS precursor NA. Thus. ZmYS1. resulting in red clover). but it was recently found that phenolics transported into the root by members of the YS/YSL family (YS1 exuded by the root in response to Fe deficiency facilitate in maize and barley. maize oligopeptide transporter (OPT) family member.1.44 ZmYS1 also transported Ni. but this alone was not able to counteract the severe and its loss results in decreased Fe uptake. however. in Oryza satiVa var.3 This resulted in a apoplastic Fe was found to decrease in response to Fe 4-fold increase in grain yield by rice grown on Fe-limited limitation. and the Fe(III)-PS complex is taken up is unclear.41 Cd disrupts Fe homeostasis in maize. the level of pathway greatly increased PS secretion. Chelation and Toxic Metals it has not been tested for Fe-phenolic transport. resulted in no decrease in apoplastic Fe. which is adapted to alkaline soils.40 this structural difference dictates substrate specificity. chelation strategy.37 Phenolics secreted by red clover roots were shown to efficiently strip Fe from purified cell walls. Indeed. the utilization of apoplastic Fe. While reduction of Fe3+ becomes transported into the root is unclear. which grows poorly on calcareous soils. in the leaf. Fe deficiency. increased. Yellow-Stripe 1 the shoot became lower. Thus. At the same time. Interestingly.3. and a constitutive chlorosis. Fe uptake from soil. perhaps via anionic channels or vesicles.39 In rice. A similar buffering role has been proposed for IRT2. while the Fe concentration in 2. ZmYS1 also serves as gateway sized from L-methionine and released from the root epidermis for a broad range of metals. mugineic acid (MA) family PSs are synthe. phenolic-mediated mobilization of apoplastic Fe deficiency response. the level of Cd uptake releases a much greater volume of PSs than most rice in maize was similar in wild-type and ys1 mutants. the decrease in Fe- Fe is an integral part of the Fe deficiency response (at least containing proteins impairs chlorophyll synthesis. and ZmYS1 has been shown to transport PS complexed with Zn. the Cd-PS complex The chelation strategy is less sensitive to pH than the is not readily transported by ZmYS1.4 although 2.44 Fe-nicotianamine (NA) transporter.

The rice. which transporter loads Fe into the xylem. No. the mutant plants even accumulated this has not yet been successfully demonstrated. suggesting that they load Fe into the seed. Interestingly. and heterologous expression in yeast indicates that it can transport Fe. Mn.50 OsYSL15 is up. although it is unclear Fe-PS uptake. the concentration of shoot maize and barley. and that it would be expressed in the same tissue as the Fe2+ tant role in Fe homeostasis. thus. surprisingly.Iron Uptake and Transport in Plants Chemical Reviews. The resulting ferric chelate reductase deficiency and had reduced Fe concentrations in their shoots. Two osysl15 interesting to determine the structural and functional sig- insertional mutants exhibited chlorotic phenotypes under Fe nificance of this change. HvIRT1 is also up-regulated under Mn deficiency and higher expression of Figure 4. 2009. 109.9 times But.52 while ectopic that will have to be engineered. This indicates a tight regulation of Fe ers OsIRT1 and OsIRT2 in the root epidermis in response homeostasis in rice and that uptake is immediately down- to Fe deficiency. and Cd in the shoot. Cu.55 including plants. in addition to near one of the four heme-coordinating sites. these Fe(II) transporters were strategy can be incorporated into grass species to augment found to be dramatically up-regulated in the roots: 30-fold Fe uptake. It is also unclear. but once in the xylem. so this may represent another step of Zn. it loads very little .51 In fact. due to its Despite its use of Fe2+ transporters. and Fe limitation in rice After entering the epidermis. in rice mutants without up.56 the gene encoding the yeast ferric chelate reductase FRE1. Fe is effluxed environment where Fe2+ is readily available. Citrate itself is transported into the 2. but the tissue-specific localization has yet to be determined. and opt3 mutants all have decreased seed Fe content. and constitutive overex- conditions.28 as in Arabidopsis.16 This is likely because interconnected cytoplasm of the root. these notransferase (NAAT) in tobacco has been shown to mutant plants died. leading to intervenial chlorosis and sterility. Fe is likely bound by increases Cd uptake and translocation. This suggests that introduction of PS synthesis into non- The elevated expression of OsIRT1 and OsIRT2 in rice grasses is feasible. Fe moves symplastically through the low ferric chelate reductase activity. Zn. for OsIRT1 and 64-fold for OsIRT2. In aerobic soils (where Fe is limiting) and in pression of the PS synthesis enzyme nicotianamine ami- hydroponic solution where only Fe3+ was supplied. expression of OsIRT1 increased Fe. the first cytosolic Fe chaperone was recently and selected for elevated activity in high pH environments. To recreate the into the xylem and moves toward the shoot through the reductase strategy system found in nongraminaceous plants. HvIRT1 is up-regulated in response to Fe deficiency. An IRT1 ortholog was recently identified in barley. indicating an impor. Fe is known to be bound by citrate. and Cd accumula- tion. identified in humans: PCBP1. has optimal activity in acidic conditions. it is premature to say whether barley transports free Fe into the root epidermis like nongrasses. When human ferritin is expressed in yeast. Fe is almost certainly efficient genotype of barley.49 soil compared with wild-type plants and produced 7. and seeds. rice roots have very potential reactivity. although it is unclear by what. Fe chelation and long-distance Fe transport. while the seed levels did not Rice compensates by expressing the divalent metal transport.47 perhaps a Ca transporter or channel or a divalent metal transporter similar to IRT1. This resulted in plants that thrived on high pH to Fe loading of seeds than Fe-PS uptake by roots. unknown chelators or chaperones (Figure 4). rice produces much less PS than greater grain yield. 10 4557 Cd primarily enters the roots through another transporter. It would be the stele. rice.52 This compensated in Similarly. making it less tolerant of calcareous soils. although these could relate more transporter. improving crops.53 This suggests these transporters have a similar range 3. and Cd. NA is an essential of the reduction strategy seen in nongraminaceous plants with part of long distance movement to the seeds. When expressed in yeast. transpiration stream. Fe increased only slightly. both OsIRT1 and OsIRT2 transport Cd. combines components transporters and an iron transport protein (ITP). Thus. once it is loaded into the seeds. Vol.48 Like AtIRT1. but we still do not understand how PSs mutants lacking PSs also resulted in increased concentrations are secreted from roots. a ubiquitously expressed RNA A common mutation in the alleles tolerant of high pH was binding protein that also facilitates Fe loading of ferritin. as mentioned above. which existence of a cytosolic Fe chaperone in plants is unproven. Of the 18 yellow-stripe like (YSL) genes in in what form the Fe is held. Long-Distance Fe Transport of substrates as AtIRT1. OsYSL15 is the primary transporter responsible for ysl1. although available. Both are similar to AtIRT1 and transport regulated once the necessary amount of Fe has been taken Fe when expressed in yeast.57 a substitution of methionine with arginine at position 312. Once HvIRT1 correlated with increased Mn uptake by a Mn transported into the root epidermal cells. coding sequence was fused with the OsIRT1 promoter to roots.54 inadequate in Fe2+-limited conditions.3.16 This also demonstrates that components of the reduction the ability to synthesize PSs. It has been proposed that NA may serve as a shuttle between the YSL Another graminaceous species. ysl3. regulated in response to Fe deficiency and is expressed on the plasma membrane in the root epidermis. increase at all. Combination of Reduction and Chelation xylem via FRD3. flowers. and developing seeds. YSLs in rice may transport Fe into the phloem. All plants more Fe in roots and shoots than wild-type under these synthesize the PS precursor NA. Zn.48 chelated. the reduction strategy alone is consume NA. Strategies where it is likely bound by nicotianamine (NA).50 Reducing OsYSL15 expression with RNAi ensure that expression of the modified gene was Fe regulated resulted in severe germination defects.49.2 At the pericycle. perhaps diffusing along many rice varieties are adapted to the anaerobic paddy the concentration gradient. Mn.16 The identified in many organisms. uptake of Fe-PS from the rhizosphere. incorporating the components of the chelation terms of Fe uptake in waterlogged soil where Fe2+ is readily strategy could increase Fe uptake in nongrass crops. was mutagenized Interestingly. Although Cu chaperones have been rice was transformed with a ferric chelate reductase.

65 While FRD3 mRNA is detected OsYSL15 and OsYSL2 are both up-regulated in response under Fe sufficiency.67 Without citrate. Ni2+. flower.62-64 of the rice YSLs also suggest a role in the long-distance indeed. Similarly.2. The eight via FRD3 (Figure 4).73 NA also readily binds Cu2+. the osfrdl1 loss of function additionally. This suggests that the as a biomarker to identify Fe-deficient plants NAS expression in rice is used to produce NA predominately The orthologue of FRD3 was recently identified in rice. Nicotianamine only a 40% decrease in citrate in the xylem.65 Despite the severe phenotype. PCBP1 is also found OsFRDL1 results in chlorotic plants with Fe precipitation complexed to ferritin in ViVo and is able to bind Fe in Vitro. rice accumulates much more NA in its roots. and reduced the Fe deficiency response to wild-type levels.76 However. 109. based on its expression utilized by the shoot. the genes most similar to HsPCBP1 mutants also resulted in increased concentrations of Zn. the concentration of complex with Fe2+.2.59 The expression pattern members also efflux citrate to mitigate aluminum toxicity. Xylem The Arabidopsis orthologs of ZmYS1 and HvYS1 do not When Fe enters the xylem. growth is tied it has a higher affinity for Fe3+ but forms a more stable to the availability of Fe. Co. while only trace . 2009. OsFRDL1 was found to transport very high in Fe-deficient rice leaves. the knockdown of PCBP1 increases cytosolic insertion mutant has increased OsIRT1 expression and Fe levels in cultured cells. because Zn2+. The loss of FRD3 results in severe chlorosis transport from root to shoot to seed via the phloem: and a constitutive Fe deficiency response. for long-distance Fe transport.70 And. have been characterized for their role in binding viral RNA Mn. and Cd in the shoot. while Perls staining showed shoot. synthesized by the condensation of three molecules results in increased IRT1 expression and Fe uptake.49.59 growth is retarded. there is 3. lamina joints.61 FRD3 is a member of the multidrug and toxic posed to transport Fe chelated by the PS precursor NA in compound extrusion (MATE) family. In the elevated expression of OsIRT1 and OsIRT2 in MA-free rice Arabidopsis genome. Thus. to increasing Fe levels. Similarly. the ferritin fills with Fe.68 It has been described as an MA.49 tolerance in Arabidopsis.72 the root vasculature. also transports Fe-PS but does not appear to be involved in Fe does not efficiently move through the xylem and is not uptake from the rhizosphere. and the loss of is coexpressed.1.75 in soil. consequently. keep increasing with increasing IRT1 sections showed that NA increases within cells in response expression. But when of S-adenosylmethionine in a reaction catalyzed by nicoti- plants increase IRT1 expression. like atfrd3. especially in the vasculature. but not Fe2+.59 In Arabidopsis. expression in oocytes showed that significant Fe3+ accumulation in the root vasculature. like OsYSL15. NA and MA levels were also While not Fe regulated. At the same time. PCBP1 was also first characterized as an the substrate specificity of uptake and translocation to the RNA binding protein.66 Xylem exudate OsYSL15 in the root vasculature. however. while barley roots accumulate (and secrete) higher levels of agement System (PiiMS). facilitating Fe loading. in the xylem. under ionomic signature for Fe deficiency69 and could be utilized both Fe sufficiency and Fe deficiency. NA is a nonproteogenic amino acid ubiquitous in higher The constitutive Fe deficiency response of the frd3 mutant plants.59 The exchange of Fe from citrate to NA has been predicted to 3. abolished Fe3+ accumulation in the base of the sheath.71 Interestingly.5. supplementation in the Purdue Ionomics Information Man. Accordingly. This unique pattern in Fe-deficient plants was further because NA is used as a precursor to MA. it may be involved in DMA-mediated Fe distribution apoplast walls. The chelators believed to bind Fe have proper. and developing collected from the top of frd3 mutant roots contained nearly seed and OsYSL2 in the phloem companion cells of the 50% less translocated Fe. all three substantiated by analyzing the shoot metal profiles of over rice NAS genes were up-regulated in the roots in response 70 000 Arabidopsis plants grown with different levels of Fe to Fe deficiency.5.1. adding citrate to the growth in reproductive organs.4558 Chemical Reviews.60 3. which is expressed in the root Arabidopsis yellow stripe like (YSL) transporters are pro- vasculature.52 This suggests that the Arabidopsis (BTR1) and regulation of flower development (HEN4 and Fe deficiency signature could be adapted to rice. Co2+. Interestingly. of which several other and out of the phloem (Figure 4). Mo concentrations decrease. The immunolabeling of NA in tomato root and shoot and Cd. indicating the requirement for a chaperone. instead it likely precipitates on the pattern. Rather. the loss of Os- In the vasculature. However. NA complexes with Fe2+ and Fe3+. The shoot concentrations of Zn. even nongrasses to grasses.61. in decreasing order of affinity. less Fe than those of wild-type ones. 7.49 OsYSL18. Co. the cytosolic Fe to ferritin. it is up-regulated 2-fold in response to to Fe deficiency and may coordinate long-distance Fe Fe deficiency. citrate is effluxed into the xylem distribution of Fe.58 and its Fe function was only shoot does not diverge between these two species or perhaps identified in 2008. Cu. suggesting a role for other citrate effluxers. and Mn2+.74 This is in contrast to barley.5.71 while OsYSL15 shoots of frd3 plants have been shown to accumulate slightly transports Fe-PS but not Fe-NA. the uptake of Zn. NA and YSLs 3. Fe is likely chelated to prevent FRDL1 reduced the concentration of Fe3+ in the xylem sap precipitation. Mn. No. Fe remains relatively constant under limitation. including delivery to the seeds.65. anamine synthase (NAS).65 The OsYSL2 transports Fe-NA but not Fe-PS. most likely via the phloem. 10 Morrissey and Guerinot Fe. while NA prevents xylem. it is believed to complex with transport Fe-PS from soil but play a significant role in the citrate. Phloem occur at pH 5.2. In the shoot. Vol. overexpression of FRD3-GFP increases aluminum transport of Fe complexes. PCBP1 likely delivers accumulates more Zn and Mn in the shoot. precipitation of Fe at the pH of phloem sap. when PCBP1 citrate when expressed in Xenopus oocytes.2. Mn.70 This suggests that there is an additional ties appropriate for their respective environments: citrate chelator besides citrate involved in Fe movement in the readily binds Fe at the xylem pH of 5. and Cd via IRT1 also goes up. where as the acidification of the rhizosphere reduces its availability NAS1 expression in roots is up-regulated by Fe deficiency. because FLK). and phloem cells at media regreened the plants.

and Ni in the shoots. resulting in decreased root Fe. Mn. Ni-NA was detected in the xylem. species for both Fe uptake and long-distance transport. yet NA began accumulating in the roots. During Fe to bind Ni. When treated with Ni. YSLs and Long-Distance Fe Transport in observed only when the quadruple mutant was created. and YSL3.83 raises the question of what chelators barley uses for long. All but YSL3 transport Fe-NA when The overexpression of NAS in tobacco and Arabidopsis expressed in yeast. which was significantly up-regulated. The double mutant also displayed movement of Fe-NA to the seeds most likely involved what was described as interveinal chlorosis.77 in part. Rice plants with disrupted PS synthesis et al. 109. Vol.2. Additionally.52 The increased seed section (section 4. exposure to Ni induces the shoots.78. The YSLs are also believed to the roots driven by the creation of new Fe sinks in the shoots. there may be a sharing of transport- Fe regulation. leads to 3.77 In the Ni hyperaccumulator Thlaspi more significant role for NA in long-distance Fe transport caerulescens.79 and overexpression of expressed in the shoots of both seedlings and adult plants. in Fe movement throughout the entire leaf. the NAS When radiolabeled Fe was applied to the cotyledons.54. This role does not appear into the interveinal tissues. NAS confers resistance to Ni. This suggests that in T.59 3. NA is translocated from the shoot to the root In Arabidopsis.81 Despite the chlorosis. and Ni appears to utilize components found in nongraminaceous rapidly accumulated in the shoots. the loss of NA.2.88 Flower and long-distance Fe transport. an Fe binding protein has been NA also plays a significant role in Ni tolerance and identified in the phloem sap of 7 day old castor bean localization. characterized.1). increasing NA levels in rice and Arabidopsis alters to the flower for loading into the developing seed. Fe accumulated only in the veins phloem of roots and shoots.77 Arabidopsis Of the eight members of the Arabidopsis YSL family.54 This suggests It is proposed that OsIRT1 transports Fe(II) into the phloem.20 in this mobilization.3.2. NA and Ni Tolerance In addition to NA. presumably because NA is mobile. based on the stele-specific up-regulation Conversely. either by loss of NAS function of signaling genes during Fe deficiency. three TcYSL family suggesting a role in Fe translocation to the shoot.82 Conversely.7.81 When NA was depleted in tobacco In rice. the high expression level of NAS genes in the in rice compared with that in barley and also a possible role shoots (relative to Arabidopsis thaliana) appears to be a key for MA in Fe translocation. suggesting NA also mediates been demonstrated to readily transport Ni-NA when ex- Fe movement to the flowers. and all were found to be expressed in increases NA levels. have been NA levels have a significant affect on metal homeostasis. growing tissues. accumulated up to 43 times more NA than wild-type plants Of particular interest is the ysl1 ysl3 double mutant. NAS2 and NAS4 were up-regulated in the root.Iron Uptake and Transport in Plants Chemical Reviews.4 in tomato and Arabidopsis or by the depletion of NA by the overexpression of the NA-consuming NAAT. OsIRT1-GUS expression was detected in the by NAAT overexpression.85-88 of Fe. to the chlorosis seen in the NA-free tomato mutant chlor- dopsis. This shoots when exposed to Fe-deficient conditions yet remained suggests a tissue-specific component of Fe sensing in chlorotic compared with wild-type plants. NA Levels and Fe Localization three members. OsIRT1 symptoms of Fe deficiency like interveinal chlorosis. nearly quadruple knock-down mutant was found to be highly all was recovered in the phloem sap associated with the 17 . was not altered. the YSLs likely play a role only been detected in the epidermis. interveinal chlorosis and sterility were 3. load Fe from senescent leaves for long-distance transport Similarly.4. No. ers and translocation pathways by Ni-NA and Fe-NA. In Arabidopsis. TcYSL3 has tive to reproductive growth.5. via the xylem sap. resulted in response to Fe deficiency. In fact. At divergence between graminaceous species and that rice the same time. NAS expression was only detected distance Fe transport.51 Expression was up-regulated of the leaf. which is discussed in the significantly more Fe than wild-type ones.77.6. YSL1.79.79 It is unclear It is often proposed that the YSLs serve to translocate Fe whether these changes are the result of greater root to shoot from the xylem into the phloem so that it can move to translocation facilitated by NA or increased metal uptake in young. especially the vasculature. caerulescens. the addition of exogenous NA. reduced growth. however. and the Ni-NA is then translocated back. indicating functional redundancy. since its expression within the root has Fe-NA transport in Arabidopsis. is a dehydrin.80 These plants interesting that Fe starvation in the interveinal areas of the accumulated significantly more Fe in both the roots and leaf is not enough to trigger the Fe deficiency response. This indicates that Arabidopsis. YSL2. Despite the varied patterns and pressed in yeast.2. all the single mutants had wild-type NA levels. and although not regulated by Ni. there are four NAS genes. orthologs.54.2. ITP 3. which in Fe-starved roots.41. an NAS overexpressing line accumulated 100 times onerVa. expression of all four NAS genes. Curie the localization of Fe. and sterility. The low levels of NA in barley component of Ni tolerance and long-distance transport. or ITP.89 The iron transport protein. A similar role was proposed for the efficient utilization of Fe in the shoot. somewhat similar OsYSL2. It is interesting that there is such a in the shoots.75 This further indicates a sensitive to Ni. in which the veins may be more important than ubiquitous NA increases Fe translocation but impairs the the interveinal tissues. In Arabi. especially in companion cells. 2009.84 Thus.88 unlike the NA-free tomato mutant. at least deficiency. Zn. root vasculature. 10 4559 amounts were detected in barley. Based on what is known about to apply to AtIRT1.59 have recently reviewed the YSLs. resulting in the increased accumulation a broad range of tissues. NAS3 members have much higher expression than their Arabidopsis expression increased 4-fold after the transition from vegeta. that NA is essential for Fe mobilization from the vasculature where it is then chelated by NA. It is and constitutive IRT1 and FRO2 expression. 3. resulting in seeds accumulating seed set were especially affected. this dramatic change in NA increased had lower Fe levels in both leaves and seeds. the Fe deficiency response as much NA as wild type.

because leaves and flower development but could not completely the flow of the xylem is driven by transpiration and seeds rescue the impaired seed set.54. The level of remobilization from especially high. respectively).4 But. The most similar genes in Arabidopsis struction. It have annotations related to stress. This figure is reproduced from ref 90 Creative Commons: Japanese Society of Plant Physiologists and Plant and Cell Physiology. At1g54410. working under the assumption that an ITP many staple crops are agronomically productive but have exists in other plant species. Consequently. 52Fe was Fe allocation to the seed. Because the plant-based diet offers relatively low amounts of bioavailable Fe. should be noted that crop breeding has often selected for regulated in response to Fe deficiency in the root (BTI1.54 Thus. (C) time course of radioactivity accumulation analyzed using PETIS. NA. YSL1.90 PETIS allows the nondestructive visual. right arrowhead +Fe). Arrowheads indicate the first detection of DC (discrimination center) (left arrowhead -Fe. while The loss or depletion of NA results in deformed flowers older leaves receive Fe from the xylem. Loading of Fe region regulates Fe distribution in barley.94 The timing and regulation of senescence leaves during flowering (+216%).54 Interestingly.93 while wheat transports 77% of shoot growth begins and accumulates significantly more Fe in the Fe to the seeds. 109. large portions of the developing 3. indicating that Fe moves quickly to the has been shown to have a significant effect on Fe accumula- phloem and nearly all is bound by ITP. often creates sinks and translocation with a positron emitting tracer imaging in the leaves rather than the seed. and Fe remobilization interact is still unclear. The images are shown at 15 and 30 min intervals (0-60 and 60-360 min.97 In response to this. and ITP remains reported 30%. 10 Morrissey and Guerinot Figure 5. Data were scored every 3 min. In tissue and intracellular level and how this ultimately affects both Fe-sufficient and Fe-deficient barley plants. while the actual movement of intake96 and are a large part of the diet in many developing Fe within the phloem occurs via ITP.1. No.81 This fits well with the observed NAS of radiolabeled 52Fe and 62Zn in rice plants. research has focused on understanding how nutrients are In barley. however. as well as significant decreases in floral Fe used to visualize and quantify the uptake and translocation accumulation.91 expression pattern in tobacco. countries. to the stele. The Arabidopsis NAS quadruple knock- shoot to seed varies by species: rice transports only 4% of down mutant also becomes chlorotic when reproductive shoot Fe to the seeds. improved grain maturation time but ignored nutrient ac- BTI2.4560 Chemical Reviews. including over 60% of all Barley children in Africa and Southeast Asia. importance of determining how Fe levels are sensed at the ization of metal movement in live plants. an Fe discrimination center (DC) in the basal transported to seeds and how this can be increased.95 How developmental changes. young leaves receive Fe primarily from the phloem. although none are specific cumulation in the grain as a desirable trait. it would appear that do not transpire. Control of Long-Distance Fe Transport in world suffer from Fe deficiency. The purified ITP tion in the seeds. transcription factors with RNAi was found to delay senes- Unfortunately.3.90 They also found 4. and several are highly up. in real time. (B) PETIS images of 52Fe accumulation after 6 h.1. serves as a shuttle. and to decrease seed Fe by over plant model organisms is difficult. found to first accumulate at a central location in the basal part of the shoot (Figure 5). obtaining large amounts of phloem sap from cence by over three weeks. the grafting of NA-depleted tobacco shoots onto 4. Realtime 52Fe movement in barley shoots: (A) gross image of Fe-deficient (left) and Fe-sufficient (right) barley analyzed using PETIS (the same frame was used for panels B and C). with highest expression being seen in flowers. Over- shoot was identified by monitoring the dynamics of Fe uptake expression of Fe-related genes. especially in anthers and pollen. most likely via the phloem. PETIS has also been and sterility. In wheat. Fe and Seeds NAS overexpressing shoots restored Fe mobilization in Fe moves to the seeds. suggesting that this 4. and YSL3 that damaging the phloem impaired Fe movement to young leaves but not old leaves. it has been proposed that NA low levels of nutrients like Fe in the seed. the .77 At the same time.92 Developing seeds receive Fe from the roots the Fe-NA requirement for normal seed development is and from senescent leaves. the knockdown of multiple NAM protein was found to preferentially bind Fe3+ but not Fe2+.16. This provides more evidence that Fe-NA is essential for flower and seed development. Vol. facilitating Fe movement in and out of Cereal seeds provide more than 50% of the world’s energy the phloem (via the YSLs). yet the Fe was only translocated to the leaves in the Fe-deficient plants. kD ITP protein.98 This shows the system (PETIS). photosystem decon- only in castor beans. 2009. At2g44060).1.

and many are expressed in the vasculature showed what were likely Fe-phytate globoids in the vacuole. the vacuolar in Arabidopsis has found that there is very little ferritin in globoids. double mutant was chlorotic. but these accumulated significantly less Fe.54. additionally. most likely due to functional redundancy. OPT3 is expressed in pollen. and most flowers did not and is expressed in the vasculature. C) three- pollen. and transiently in the ER. 2009. since Perls staining of Fe in embryos101 essential role for AtOPT3 in seed development. however. suggesting compensation. the resulting In Arabidopsis.2. indicating that Fe was not being mobilized from 4. provascular strands of the embryo in wild-type seeds. indicating seeds. because Fe-NA is critical in seed development. while IRT1 expression in the flower increased. An opt3 strongly resembles the vascular localization of Fe demon- knock down line. These phenotypes are similar to the floral deformity seeds. in response to Fe deficiency.1. YSL1 Figure 6.105 This and the AtYSLs. in addition to expression in the flower. while in the Vit1 mutant. No. plays an essential role in Fe loading of mislocalization of Fe resulted in decreased seedling viability the seed. Copyright 2006 The American lines contained 30-65% less Fe and germinated more slowly Association for the Advancement of Science. and reproductive organs.87 This suggests a role dimensional rendering of Fe KR X-ray fluorescence in Col-0 and in Fe loading from senescent leaves for transport to develop. resulting in brown necrotic spots. in the phenotypes. have reported which disappeared as germination progressed. Visualization of Fe distribution by synchrotron X-ray seed development requires not just the availability of NA fluorescence microtomography106 showed Fe concentrated in but also specific Fe-NA transporters. (B. young siliques. respectively. It should also be noted that IRT1 expression in the flower is also exclusively in the anther. Watering plants with exogenous Fe could not restore Fe accumulation in the seeds. especially within the seed: (A) three-dimensional rendering of total X-ray in senescent leaves. Fe was not associated with the vascular 4.20 indicating a role for both Fe-NA and Fe(II) in flower development. VIT1 transports Fe2+ into the vacuole. absorption of a wild-type Arabidopsis seed. Thus. as had been assumed that found as much as 90% of Fe in ferritin.100 double nramp3 nramp4 mutant. but rather was seen throughout the hypocotyl and radicle and was concentrated in a layer of cells just inside OPT3. OPT3 system. Vol. the nramp3 nramp4 double mutant OPT3 is unknown.105 While the loss and irregular and 80% less likely to germinate than wild-type overexpression of VIT1 does not affect total Fe levels in seeds. none. on Fe-deficient medium. The expres- sion pattern of YSL3 is somewhat similar to that of YSL1: 4.108 appear . Fe is severely mislocalized in the loss of function and sterility seen in plants lacking NA. vacuolar Fe loading via VIT1 is vasculature.99.101 The opt3-2 plants also exhibit constitutive expression of genes 4. compensated by other transporters or chelators. When a second quadruple mutant with NA synthesis completely abolished was created. Indeed. role in Fe-NA delivery to the developing seed. which are ions complexed with phytate. The Fe stored in the vacuoles of the vasculature may be in the opt3 null mutant is embryo lethal.100 Unlike the ysl mutants. This was based on earlier experiments in legumes in Arabidopsis seeds is not ferritin.2. Loss of the VIT1 transporter changes Fe distribution expression was found in and around leaf veins.105 seeds. essential for proper Fe distribution in the embryo. especially during produce siliques.103 Recent work based on earlier work in legume seeds. the silique on Fe-limited soil. indicating an the Fe3+ form. if not essential. In Arabidopsis. Vit1-1. This leads to the accumulation of very In Arabidopsis. But. 10 4561 level of seed Fe only decreased 46%. the seeds of the ysl1 loss of function mutant with permission from ref 105.Iron Uptake and Transport in Plants Chemical Reviews. the YSLs play an important. NRAMP3 and NRAMP4 involved in the root Fe deficiency response. Thus. causing the germination defects seen on Fe- limited media. and embryo. Interestingly.81 indicating that mutant.107 chelator. Reprinted ing seed. and the developing embryo. a member of the family that includes ZmYS1 the abaxial epidermis of the cotyledons (Figure 6). opt3-2. allowed embryo formation in strated by SXRF. hence the germination phe- storage state of Fe in Arabidopsis seeds was unknown. regardless of exogenous Fe supply. to the vacuoles in the vasculature but transport Fe out of especially during the seed-filling stage. but its phenotypes and relation to the seeds contain the same level of Fe as wild-type but YSLs suggests it likely transports chelated Fe or an Fe produce seedlings that grow poorly on Fe-limited soil.2. The few resulting seeds were small and embryo and seed development. The substrate of the vacuole. Like Vit1. VIT1 in the vasculature of shoots and in pollen and anthers. the result was sterility. it has been observed that developing seeds not expressed until the third day of germination. which in expression in the root and shoot vasculature is up-regulated turn determines seedling viability under low Fe conditions. the first store Mn and Zn complexed with phytate in the vacuoles of two days of growth rely on mobilization of vacuolar Fe stores the embryo and endosperm. although it was long assumed to be stored in ferritin in the this also demonstrated that the primary storage form of Fe plastid. There are eight other members of the Arabidopsis Visualization of wild-type seeds by electron microscopy OPT subfamily. NRAMP3 and NRAMP4 also localize high levels of Fe in leaves. Instead.104 raising the possibility that in Arabidopsis most seed that YSL1 plays a role in seed loading that cannot be Fe is bound by phytate or some other chelator in the vacuole.102 The via NRAMP3 and NRAMP4. with both seeds identically oriented.88 When the two loss of function mutants were crossed.2. 109.2. notypes of Vit1 and nramp3 nramp4 mutants.1. Because the Fe uptake transporter IRT1 is In Arabidopsis. the globules remained during germination. Storage of Fe the vacuole.

116 Thus. while increasing the amount of ferritin. rice. In the mitochondria. including in the maize ectopically express Aspergillus phytase and soybean ferritin embryo and the aleurone cells of wheat.122 Of course.119-121 rats fed ferritin overexpressing rice recov- In addition to vacuoles.3. Early attempts to reduce phytic acid across the whole plant successfully reduced accumulation in seeds. Fe is of phytate synthesis. the loss of sequestered by ferritin and frataxin (FH). Because phytate represents around 1-2% of the dry weight of cereal seeds. as was fed to cultured human cells. promoters produced the same fold increase in Fe as trans- ferritin likely serves more as an Fe buffer. No. invasion.g. overexpression of ferritin does have only FER2 is expressed in developing seeds. which membrane the transporter localizes or even what it tered in ferritin. but these plants often germinated poorly and were more susceptible Figure 7. suggesting further Fe to prevent oxidative stress. ferritin has been nent of Fe storage in seeds.2. most likely to minimize the phytate precursors did alter phosphate sensing. possibly the inner silencing of an ABC transporter.113 this poses a serious impediment to dietary Fe uptake. ferritin in tobacco resulted in a constitutive Fe deficiency regulated in response to the plant hormone abscisic acid response. most notably during Fe deficiency and germination. a combination of the two approaches has been Large amounts of the antinutrient phytate accumulate in undertaken. several strategies have been employed to reduce the amount of phytate in seeds. at the same time. the increase in sequestered Fe fact. In the developing world. the flowers accumulated more Fe and were highly resistance to Fe overload. The oxidative stress. The fer2 loss of function mutant does not affect Fe also a 2-fold increase in Cd when grown on contaminated accumulation or seed viability under normal conditions. Fe(III) is reduced by FRO7. the overexpression of soybean and bean 4. causing greater Fe uptake and accumulation but (ABA).118 Although it is unclear to membrane-localized PIC1. Fe Bioavailability for Humans Finally. Fe is seques.109 While these muta- NRAMP4 transport Fe out of the vacuole. NRAMP3 and produce seeds with 93% less phytate. it is believed that the Fe NRAMP4 are essential for seed viability under Fe limited complexed in ferritin is readily absorbed and a highly conditions and implicate the vacuole as an integral compo. both loading of Fe into the seed vacuole via VIT1 Although the mechanism of dietary ferritin uptake in the and its release during germination via NRAMP3 and human gut is unknown. It reduction of phytate in seeds affects Fe homeostasis has not would thus be interesting to determine the Fe storage form been examined. in soil. and pathogen sensitive to Fe supplementation. including Fe. Vol.117 Recently in Arabidopsis. This resulted in deformed. Zn. the prevalence of phytate in the plant-based diet is believed to contribute to the high rate of Fe deficiency and anemia. accessible source of Fe. A second approach is to overexpress ferritin in seeds.111 in the endosperm.125 4. Thus. the disruption of the transported into the vacuole by FPN2 and VIT1 (although they inositol polyphosphate kinases required for the later steps are expressed in different tissues). NA) mutants. One obvious approach is to disrupt phytate biosynthesis.116 Consequently. Indeed. ferritin was estimated to account for only 5% of total produced improved resistance to oxidative stress. Fe stored within ferritin is believed to be safe and have high bioavailability. Overexpression of soybean germination. high fiber diets have been shown to induce Fe deficiency in healthy women. Fe uptake was significantly . its loss prevents phytate from accumulating in the seed without compromising seed viability.115 because phytate is presum- ably binding Fe from other foods in the intestine.114 In fact. indicating that the Fe is bioavail- with ferritin. 109. Accordingly.110 Accordingly. Intracellular Fe transport and sequestration. is able to bind Fe in the vacuole. but it would be interesting to look at the in the reduced phytate Arabidopsis mutant.126 This increased total Fe content in seeds Phytate is composed of a phosphorylated myo-inositol ring by 20-70% and resulted in the degradation of nearly all and strongly chelates metal cations.112 these salts accumulate as globules in the vacuole.124 Ferritin overexpression with more powerful less functional flowers and increased oxidative stress. Transgenic maize plants were generated to the seeds of many staple crops. FH also plays a role in Fe-S cluster assembly authors noted that this could be overcome by using promoters or repair. 2009. Fe is found in the plastids bound ered from Fe deficiency.109 Perhaps ferritin interplay between vacuolar and plastid Fe pools in these levels increase to compensate or another chelator (e. 10 Morrissey and Guerinot mentioned above.. AtIPK1 and AtIPK2. When paste from the resulting seeds Mn. and during consequences for the plant.3. low phytate maize the matrix. was found to known to be complexed with phytate and NA.4562 Chemical Reviews. FER2 ferritins in rice seed resulted in 2-3-fold increases in seed Fe content. Within the vacuole. sequestering free genics with weaker promoter constructs.123 Over- seed Fe. Conversely. Arabidopsis has four ferritin genes. and soybean seeds were generated by the seed-specific and then taken up by an Fe(II) transporter. transports. Fe is to stress. increasing Fe accumulation is limited by Fe uptake and transport and not by ferritin levels. viewed as a means of increasing bioavailable Fe in staple crops. and endogenous phytate.104 When the other three ferritin genes were knocked expression of alfalfa ferritin in tobacco produced an increased out. and barley. of which able. Thus. FER2 is the only ferritin up. How the to be a primary Fe storage form in Arabidopsis seeds. ATM3 is believed to transport Fe-S clusters out of specific for the seeds. Within the chloroplast. To enter the chloroplast. oxidative stress. making it unavailable for uptake.98. Fe is tions did not affect seed yield or germination.

which correlates with the low levels Plant mitochondria require Fe for respiration. Mitochondria when Fe is limiting. and the synthesis of Fe-S clusters. the Fe-binding proteins ferritin and frataxin have suggested that PIC1 transports Fe and Cu across the been localized to the mitochondria in several organisms. integral to the Fe sensing mechanism. This implies that the plastid Fe pool in seeds is insubstantial.110 Additionally. resulting in to the chloroplast in the ferric form and must be reduced increased oxidative stress and deformed flowers. The roles of the ferritin paralogs is differentiated by heme biosynthesis. rather than the plastid. This mislocalization of Fe also changes the on its putative transit peptide. all of which localization and regulation: FER2 is only expressed in the take place in the chloroplast. homeostasis within the chloroplast. such as NtNAS. and both transporters and Fe sequestering proteins have been found to be essential for mitochondria function (Figure 7). the FRO7.104 This further impli.2. yet very little is known about seeds. Thus.107 the fro7 loss of function mutant accumulating in the flower. because translocon component. proper Fe homeostasis in the mitochondria is vital. Fe-regulated genes in the shoot localize to the mitochondria. FER1. but also in the whole cell. The triple facilitated import of ferrous Fe into the plastid is essential mutant also showed no decrease in photosynthesis. 10 4563 higher compared with those fed wild-type seed paste.132 and is essential for Fe homeostasis within 5. the chloroplasts of the fro7 mutant contain 33% although it is unclear whether this is by physically sequestering less Fe. While the ing that ferritins are not essential for chloroplast development nramp3 nramp4 double mutant grows poorly when germi. These chondria.131 However. Intracellular Fe 5. AtYSL1. while the other three ferritins are expressed in the how Fe is transported in and out of this organelle.133 but cates the vacuole.Iron Uptake and Transport in Plants Chemical Reviews. PIC1 localizes to the inner envelope of the chloroplast131. the ferritins prevent excess free Fe from nated on Fe-limited soil. is highly dependent on energy from the mitochondria. This sup- to enter the chloroplast. and the expression of the root Fe uptake transporter the chloroplast (Aramemnon Plant Membrane Database). whereas the outside the plastid.1. of the chloroplast affects the Fe homeostasis of the entire attempts to increase bioavailable Fe in seeds are becoming cell. biosynthesis.3.139 They appear to play a very change in the pic1 mutant. the expression of the three nonseed its levels within the plastid in response to cellular demand ferritins increases in response to high Fe levels.54. the plants are dwarfed and important role in metal homeostasis not only in the mito- chlorotic. regulates long-distance Fe transport to the flower. other than confirming their presence in being utilized properly in the plastid and instead accumulates purified mitochondria from pea and Arabidopsis. rather than protein AtOPT3. This indicates that the chloroplast is The fer4 loss of function mutant does not have a phenotype. and Fe-S cluster assembly. Although overall Fe levels in the leaf do not including Arabidopsis. FER4 is predicted to localize to It is believed that chloroplasts hold nearly 90% of the Fe the mitochondria. as the primary site the combination of electrons and free Fe is highly toxic.1. although it may also target to cells.88. both Fe mobilization from the vacuole and Fe import into the plastid are essential for seedling development 5. or function. or be dual targeted to both organelles.129. Plastids to localize to the plastid. especially microsporogenesis. FRO7 excess free Fe to prevent oxidative stress.20. 5. 5. Ferritin In Arabidopsis.110 within a leaf. dies. heme of ferritin found in Arabidopsis seed. regulating roots.100 translocation.104 indicat- for seedling growth under Fe-limited conditions. The expression of the ferric reductase FRO7 on the the triple mutant showed a shift in Fe accumulation from chloroplast membrane indicates that some Fe is trafficked stem to flower when supplemented with Fe. Accordingly. This prevents excess Fe movement to the flower. IRT1 was repressed.1. PIC1 Flower development.128 Previous experiments with ports the hypothesis that chloroplasts are an important Fe purified pea chloroplasts showed that Fe was likely trans. resulting in decreased photosynthetic efficiency and Fe in the shoot or whether the Fe status of the plastid fewer healthy photosystems. AtYSL3. Fe is required for photosynthesis. Transport. where it causes damage. shoots and flowers. with impaired chloroplast development. genes in the root.110 Ferritins appear to buffer Fe levels and sequester 5.1.127 Indeed. in addition to the expression of Fe deficiency response more successful.136-138 membrane. Ferritin and Frataxin the plastid and plant as a whole. and FER3 are predicted 5.2. and AtIRT1. AtFER4 is the most likely to expression of nonplastid. FER2. Instead. seed FER2 is expressed in response to the plant hormone ABA. in addition to FER1 expression in the ers likely serve as the gateway for Fe (Figure 7).132 The heterologous expression of the plastid-localized transporter in yeast Recently. 2009. because the Fe status perhaps because one or more of its paralogs are also targeted .134 Maintaining PIC1 (Tic21) was originally identified as a chloroplast mitochondrial Fe levels is thus of high importance.2. Thus.1.1.104 When the three genes encoding nonseed ferritins were knocked out.139 Based in ferritin. 109.130 plastids. during germination. sink and that ferritins may sequester some Fe in the leaf ported across the inner envelope in the ferrous state. many Fe-related it has also been proposed that developmental defects seen genes are highly expressed in the anthers (the portion of the in the loss of function mutant are related to impaired Fe male organ of the flower containing pollen). Vol. because it immunoprecipitates with the Fe deficiency produces deformed mitochondria in rice pollen major components of the Toc and Tic translocon. this suggests that Fe was no longer ferritins in plants. plastids were also found to have elevated levels of ferritin Very little research has been done on mitochondrial and lacked thylakoids. and reduces seed yield.135 Appropriately. No.128 Additionally. for Fe storage in seeds and subsequent site for mobilization Thus.

and its tion of Fe shifts to the chloroplast and FER1 expression loss is embryo lethal. It database68 has allowed the identification of the Fe-deficiency would be very interesting to investigate how the constitutive signature and will likely yield more unexpected discoveries export of Fe-S clusters from the mitochondria affects the in the future. the expressed in the vasculature. which is then released into the AtATM3 was able to rescue the ∆atm1 yeast. and the atm3 dwarfs were more sensitive to Cd clarifying ferritin function and in identifying the vacuole’s than wild-type plants.143. similar vacuolar globules were identified as containing Fe and phosphate.3.3. Thus. but as studies of rice and other that IRT1 expression may be up-regulated. Vol.2. Conclusions and Future Directions ATM3 also appears to play a role in Cd detoxification in Much progress has been made in the past few years in Arabidopsis. Interestingly. 10 Morrissey and Guerinot to the mitochondria or frataxin is able to compensate. in response to Fe deficiency. NA and the Vacuole serve as a chaperone. 2009. mediating Fe delivery to the Fe-S cluster assembly scaffold. it terization of transcriptional changes in specific root layers has been suggested that ATM3 may also function to export of Arabidopsis has proven an invaluable resource. in addition to ing the resolution of our understanding of the Fe deficiency its role in transporting Fe-S clusters. it is worth speculating In Arabidopsis. storing and releasing Fe flies.140. In addition to sequestering Fe.2.144 Frataxin is essential to growth increases. 6. the vacuole serves as the most overload. while ATM3 overexpression enhanced role in Fe storage and mobilization during seed set and Cd resistance.136.148 Like ∆atm1 yeast. these vacuolar Fe globules were only seen in Like FER4. frataxin is believed to 5.141 Interestingly. vacuolar membrane in the vasculature of the roots and shoots chondria of these plants accumulated more nonheme. because FER4 ex. 109. Because ATM3 is closely related to the fission germination. however. but the thousands of mutant Arabidopsis lines into the PiiMS concentration of Fe and other metals were not reported. because line also accumulates more Fe in the root. VIT1 is increased oxidative stress. The increase in shoot Cd suggests generalized to all plants. could and NRAMP4.147 The Arabidopsis ATMs were first identified by the chlorotic. increas- chelated Cd complexes out the mitochondria.105 and like NRAMP3 and NRAMP4. Vacuole likely. While it is expressed in response to Fe As described earlier. especially in Cd or Pb. VIT1. the only identified mitochondrial Fe whether the depletion of NA shifts Fe sequestration within transporters are the ATMs. and the expression they were found in the vacuole proteome of Arabidopsis of FER1 and FER4 is increased. Some of the discoveries in Arabidopsis can be Fe deficiency response. electron microscopy cluster assembly. protecting is essential for functional Fe-S clusters and that even the cell from oxidative stress. presumably to prevent suspension cells. VIT1 most likely loads Fe into protein Fe than those of wild-type plants.145 The knock-down of frataxin in NA is found in the vacuole74 and is likely chelated to Fe Arabidopsis is not lethal but results in increased ROS and there. of chlorotic leaf sections showed the appearance of electron dense globules in the vacuole.4564 Chemical Reviews. but only filling the vacuole with Fe. in human cells has been linked to cytosolic Fe depletion. When expressed in yeast. Additionally. it was recently demonstrated that phosphorus pression was highest in the flowers and floral stalk. leaf sections from plants grown under phosphorus sufficiency plants. No. grasses have shown. consequences in terms of sequestration of free Fe and Fe-S When NA was depleted in tobacco. suggesting that ferritin is now binding Fe.149 These cells of the vasculature during Fe deficiency by NRAMP3 results suggest that ATM3. the recent large-scale charac- yeast SpHMT1. ATM3 analyzed.2. The vacuole also stress. because the overexpression of mitochondrial ferritin showed Fe and phosphate in globules in the vacuole. perform a similar function of Fe-S cluster export from the Arabidopsis mitochondria. frataxin is not Fe regulated.74 It has been reduced activity.146 Interestingly. resulting in the vacuole. Fe-S cluster-related genes was found primarily in the cytoplasm during Fe deficiency in the mitochondria are up-regulated in the mutant.1. under phosphorus deficiency. and NRAMP4 clusters from the matrix. the localiza- mitochondrial ferritin.3. non.137 FER4 appears to play an important role in the (Figure 7) in response to changes in cytosolic Fe levels. and Fe sufficiency but was shown to concentrate in the the resultant Fe-S-containing proteins (like aconitase) have cytoplasm and vacuole during Fe overload.150 ATM3 is up-regulated in roots treated with studying Fe homeostasis in Arabidopsis.151 Thus. the deposition of ICP-MS data from ATM3 also accumulate more Cd in roots and shoot.152 In the shoots of tomatoes and peas. YSL4 and YSL6 are candidates. are orthologs of ScATM1.142 in addition to the developing embryo.110 Like the mitochondrial ferritin in humans and fruit plays a role in the roots and shoots. mitochondria-rich reproductive organs. if not the other ATMs.144 The knock-down out of the vacuole.54 Although these were not 5. the role of VIT1 may be Arabidopsis ATMs localized to the mitochondria. NA oxidative stress. frataxin is expressed in the mitochondria of the the cells surrounding the vasculature and not in other tissue flowers. FER4 is not essential for mitochondria function under normal conditions.73. because it has functions beyond mitochondrial Fe sequestra- tion. More 5.74 and this excess Fe-NA decreased expression of frataxin has serious phenotypic may then be sequestered in the vacuole. it is down-regulated in response to oxidative important Fe store in Arabidopsis seeds.148. there are species-specific aspects of Fe . ScATM1 localizes to the mito- chondrial inner membrane and is believed to efflux Fe-S 5. mito. It would availability controls the subcellular localization of Fe in be interesting to see the effects of FER4 overexpression in Arabidopsis leaves. If Fe-NA complexes are indeed transported in and decreased vegetative growth and seed set. Similarly. NRAMP3. half-molecule ABC proteins that the vacuole into phytate complexes. However.144 This indicates that Arabidopsis frataxin proposed that NA functions as an Fe(II) scavenger. a vacuolar phytochelatin-Cd transporter. dwarf phenotype of the atm3 loss AtNRAMP3 and AtNRAMP4 are both expressed on the of function mutant (or sta1).143 Unlike layers.151 Using X-probe microscopy. Plants overexpressing response.

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