Jean-François Briat

Involvement of NRAMP1 from Arabidopsis thaliana in iron transport.

Nramp genes code for a widely distributed class of proteins involved in a variety of processes, ranging from the control of susceptibility to bacterial infection in mammalian cells and taste behaviour in Drosophila to manganese uptake in yeast. Some of the NRAMP proteins in mammals and in yeast are capable of transporting metal ions, including iron. In plants, iron transport was shown to require a reduction/Fe(II) transport system. In Arabidopsis thaliana this process involves the IRT1 and Fro2 genes. Here we report the sequence of five NRAMP proteins from A. thaliana. Sequence comparison suggests that there are two classes of NRAMP proteins in plants: A. thaliana (At) NRAMP1 and Oriza sativa (Os) NRAMP1 and 3 (two rice isologues) represent one class, and AtNRAMP2-5 and OsNRAMP2 the other. AtNramp1 and OsNramp1 are able to complement the fet3fet4 yeast mutant defective both in low- and high-affinity iron transports, whereas AtNramp2 and OsNramp2 fail to do so. In addition, AtNramp1 transcript, but not AtNramp2 transcript, accumulates in response to iron deficiency in roots but not in leaves. Finally, overexpression of AtNramp1 in transgenic A. thaliana plants leads to an increase in plant resistance to toxic iron concentration. Taken together, these results demonstrate that AtNramp1 participates in the control of iron homoeostasis in plants.

Plant responses to metal toxicity.

Metal toxicity for living organisms involves oxidative and/or genotoxic mechanisms. Plant protection against metal toxicity occurs, at least in part, through control of root metal uptake and of long distance metal transport. Inside cells, proteins such as ferritins and metallothioneins, and glutathion-derived peptides named phytochelatins, participate in excess metal storage and detoxification. Low molecular weight organic molecules, mainly organic acids and amino acids and their derivatives, also play an important role in plant metal homeostasis. When these systems are overloaded, oxidative stress defense mechanisms are activated. Molecular and cellular knowledge of these processes will be necessary to improve plant metal resistance. Occurrence of naturally tolerant plants which hyperaccumulate metals provides helpful tools for this research.

Cytokinins negatively regulate the root iron uptake machinery in Arabidopsis through a growth-dependent pathway.

Plants display a number of biochemical and developmental responses to low iron availability in order to increase iron uptake from the soil. The ferric-chelate reductase FRO2 and the ferrous iron transporter IRT1 control iron entry from the soil into the root epidermis. In Arabidopsis, expression of IRT1 and FRO2 is tightly controlled to maintain iron homeostasis, and involves local and long-distance signals, as well as transcriptional and post-transcriptional events. FIT encodes a putative basic helix-loop-helix (bHLH) transcription factor that regulates iron uptake responses in Arabidopsis. Here, we uncover a new regulation of the root iron uptake genes. We show that IRT1, FRO2 and FIT are repressed by the exogenous addition of cytokinins (CKs), and that this repression acts at the level of transcript accumulation, and depends on the AHK3 and CRE1 CK receptors. The CKs and iron-deficiency signals act through distinct pathways to regulate the soil iron uptake genes, as (i) CK repression is independent of the iron status, (ii) IRT1 and FRO2 downregulation is unchanged in a fit loss-of-function mutant, indicating that FIT does not mediate CK repression, and (iii) the iron-regulated genes AtNRAMP3 and AtNRAMP4 are not downregulated by CKs. We show that root growth-inhibitory conditions, such as abiotic stresses (mannitol, NaCl) and hormonal treatments (auxin, abscissic acid), repress the iron starvation response genes. We propose that CKs control the root iron uptake machinery through a root growth dependent pathway in order to adapt nutrient uptake to the demand of the plant.

Chapter 4 Iron Dynamics in the Rhizosphere: Consequences for Plant Health and Nutrition

Abstract Iron is an essential micronutrient for most organisms due to its role in fundamental metabolic processes. In cultivated soils, soil solution iron is mostly oxidized [Fe(III) species] unless local anoxic conditions develop. The concentration of these Fe(III) species is small in soil solution due to the low solubility of ferric oxides, oxyhydroxides, and hydroxides, which is minimal at neutral and alkaline pH. In the rhizosphere, iron concentration in the soil solution is even lower because of its uptake by aerobic organisms (plants and microorganisms), leading to a high level of competition for Fe(III). In order to face iron competition, these organisms have evolved active uptake strategies based on acidification, chelation, and/or reduction processes. Iron competition plays a major role in microbial and plant–microbe interactions in the rhizosphere. This review summarizes current knowledge on the iron status in soils and rhizospheres, and the acquisition strategies of plants and microbes. This review also shows how the dynamic interactions between soil minerals, plants, and microorganisms impact plant health and nutrition. Analysis of these complex interactions offers an interesting case study of research on rhizosphere ecology integrating different scientific expertises and approaches.

The iron-sulfur cluster assembly machineries in plants: current knowledge and open questions

Many metabolic pathways and cellular processes occurring in most sub-cellular compartments depend on the functioning of iron-sulfur (Fe-S) proteins, whose cofactors are assembled through dedicated protein machineries. Recent advances have been made in the knowledge of the functions of individual components through a combination of genetic, biochemical and structural approaches, primarily in prokaryotes and non-plant eukaryotes. Whereas most of the components of these machineries are conserved between kingdoms, their complexity is likely increased in plants owing to the presence of additional assembly proteins and to the existence of expanded families for several assembly proteins. This review focuses on the new actors discovered in the past few years, such as glutaredoxin, BOLA and NEET proteins as well as MIP18, MMS19, TAH18, DRE2 for the cytosolic machinery, which are integrated into a model for the plant Fe-S cluster biogenesis systems. It also discusses a few issues currently subjected to an intense debate such as the role of the mitochondrial frataxin and of glutaredoxins, the functional separation between scaffold, carrier and iron-delivery proteins and the crosstalk existing between different organelles.

Post-Translational Regulation of AtFER2 Ferritin in Response to Intracellular Iron Trafficking during Fruit Development in Arabidopsis

Ferritins are major players in plant iron homeostasis. Surprisingly, their overexpression in transgenic plants led only to a moderate increase in seed iron content, suggesting the existence of control checkpoints for iron loading and storage in seeds. This work reports the identification of two of these checkpoints. First, measurement of seed metal content during fruit development in Arabidopsis thaliana reveals a similar dynamic of loading for Fe, Mn, Cu, and Zn. The step controlling metal loading into the seed occurs by the regulation of transport from the hull to the seed. Second, metal loading and ferritin abundance were monitored in different genetic backgrounds affected in vacuolar iron transport (AtVIT1, AtNRAMP3, AtNRAMP4) or plastid iron storage (AtFER1 to 4). This approach revealed (1) a post-translational regulation of ferritin accumulation in seeds, and (2) that ferritin stability depends on the balance of iron allocation between vacuoles and plastids. Thus, the success of ferritin overexpression strategies for iron biofortification, a promising approach to reduce iron-deficiency anemia in developing countries, would strongly benefit from the identification and engineering of mechanisms enabling the translocation of high amounts of iron into seed plastids.

Chapter 12 Role of Iron in Plant–Microbe Interactions

Abstract Iron is an essential micronutrient for plants and associated microorganisms. Iron nutrition of these organisms relies on the soil supply. However, bioavailability of iron in cultivated soils is low. Plants and microorganisms have thus evolved active strategies of iron uptake based on acidification, chelation, and/or reduction processes. Iron acquisition by these organisms leads to complex interactions ranging from mutualism to competition. In the rhizosphere, plants support abundant and active microbial communities through the release of rhizodeposits. Iron uptake by these microorganisms and by the host plant decrease even more the concentration of iron in solution. Therefore, there is an intense competition for iron among rhizosphere microorganisms, favoring those with the most efficient iron uptake strategy. This is the case for fluorescent Pseudomonas bacteria that synthesize siderophores, called pyoverdines or pseudobactines, which have a high affinity for iron and suppress fungal phytopathogens and deleterious microorganisms. Pyoverdines also elicit plant defense reactions and contribute to plant iron acquisition. Taken together, these mutual effects promote plant growth and health. However, competition for iron may also occur between plants and microbes during pathogenesis. Siderophores contribute to the iron uptake of the host plant and to the virulence of pathogens; conversely, host plants activate mechanisms aimed at depriving pathogens of nutritional iron. The iron-withholding mechanisms of the host plant rely on controlling its iron homeostasis. In this chapter, we describe the strategies of iron uptake of plants and microorganisms, the resulting complex interactions between them, and the challenges represented by their monitoring in agroecology.

Metal resistance in yeast mediated by the expression of a maize 20S proteasome α subunit

Transformation of yeast cells with a maize cDNA ZmPAA, encoding a 20S proteasome α-subunit, conferred resistance to nickel, cadmium and cobalt. This resistance is not linked to a modification of the intracellular nickel content, as no accumulation of nickel was measured between yeast cells transformed with a void vector or the ZmPAA cDNA. The abundance of the ZmPAA mRNA was increased in the shoots of maize plants upon nickel treatment. These results suggest that the proteasome might be involved in nickel resistance by scavenging metal oxidized proteins both in plants and yeast.

The soil type affects both the differential accumulation of iron between wild-type and ferritin over-expressor tobacco plants and the sensitivity of their rhizosphere bacterioflora to iron stress

Transgenic tobacco P6 over-expressing ferritin is known to activate iron transport systems and to have increased iron content. Iron phytoextraction by this transgene is then expected to be higher than that of the wild-type (WT). In the present study, the possibility to modify iron availability for bacteria via the cultivation of the transgene P6 was explored by comparing the sensitivity to iron stress of bacteria isolated from the rhizosphere of the two plant genotypes (WT and P6). This sensitivity was evaluated by measuring the bacterial density when plated on a solid media depleted (supplemented with 8-hydroxiquinoline) or not (supplemented with Fe-8-hydroxyquinoline) in iron. The experimental conditions favorable to the differential iron accumulation between the wild-type and transgenic tobacco were identified. The two plant genotypes were grown in three soils (Hervau, Thory and Oudun) chosen for their differences in iron content, and the plants were yielded at three stages (vegetative, floral bud and flowering). The highest differential accumulation of iron in favor of the over-expressing transgene was found in the plants at the floral bud stage when cultivated in the Oudun and Thory soils. Since at that stage, the plant growth was significantly higher in the Oudun soil, the phytoextraction of iron was the highest in this soil. At the floral bud stage, bacteria isolated from the rhizosphere of the transgene cultivated in the Oudun and Thory soils appeared to be less susceptible to iron stress than those from the wild-type. Bacterial density recovered on agar medium depleted in iron was significantly the highest in the rhizosphere of the transgene cultivated in the Oudun soil. Altogether, these data indicate that the over-expressing ferritin transgenic plants, that accumulate and extract more iron from the rhizosphere than the wild-type plants, select in their rhizosphere bacteria less susceptible to iron stress compared to those selected by the wild-type plants.

Iron around the clock

Carbon assimilation, a key determinant of plant biomass production, is under circadian regulation. Light and temperature are major inputs of the plant clock that control various daily rhythms. Such rhythms confer adaptive advantages to the organisms by adjusting their metabolism in anticipation of environmental fluctuations. The relationship between the circadian clock and nutrition extends far beyond the regulation of carbon assimilation as mineral nutrition, and specially iron homeostasis, is regulated through this mechanism. Conversely, iron status was identified as a new and important input regulating the central oscillator, raising the question of the nature of the Fe-dependent signal that modulates the period of the circadian clock. Several lines of evidence strongly suggest that fully developed and functional chloroplasts as well as early light signalling events, involving phytochromes, are essential to couple the clock to Fe responses. Nevertheless, the exact nature of the signal, which most probably involves unknown or not yet fully characterized elements of the chloroplast-to-nucleus retrograde signalling pathway, remains to be identified. Finally, this regulation may also involves epigenetic components.