Yellow Stripe-like Research Articles

Dynamic Expressions of Yellow Stripe-Like (YSL) Genes During Pod Development Shed Light on Associations with Iron Distribution in Phaseolus vulgaris.

The global prevalence of iron deficiency-induced "hidden hunger" highlights a critical health concern, underscoring the pressing need to improve iron nutrition through safe and efficient means, such as increasing iron intake from plant-based foods. Yellow Stripe-Like (YSL) genes play a crucial role in long-distance iron transport between source and sink tissues in plants. Here, we report on the analysis of YSL family genes in the common bean (Phaseolus vulgaris L.), an iron-rich legume crop. We identified 9 YSL genes in the common bean genome using BLAST and HMM methods. Gene duplication analysis revealed that PvYSL7a and PvYSL7b originated through tandem duplication events. Structural analysis noted an absence of conservative motifs in PvYSL3b and PvYSL7a, which led to distinct predicted 3D protein structures. Leveraging publicly available RNA-seq data from developing bean pods, the expression patterns of PvYSL genes alongside pod and seed development were analyzed. Notably, PvYSL7a and PvYSL7b, as well as PvYSL1a and PvYSL1b, exhibited diverged expression patterns in seeds, signifying their functional divergence in this tissue. Moreover, PvYSL3a and PvYSL3b exhibited divergent expression patterns in both pod walls and seeds during pod development, underscoring their distinct roles in facilitating iron transportation between pods and seeds. This study provides valuable insights into the gene regulatory basis of iron accumulation in bean pods and seeds.

A mutation in the promoter of the yellow stripe-like transporter gene in cucumber results in a yellow cotyledon phenotype

Leaf color mutants in higher plants are considered to be ideal materials for studying the chlorophyll biosynthesis, photosynthesis mechanism and chloroplast development. Herein, we identified a spontaneous mutant, yc412, in cultivated cucumber that exhibited yellow cotyledons. The yellow-lethal mutant was diagnosed with an abnormal chloroplast ultrastructure, and reduced photosynthetic capacity and pigment content. Through bulked segregant analysis-based whole-genome sequencing and fine genetic mapping, we narrowed the yellow cotyledons (yc) locus to a 96.8 kb interval on chromosome 3. By resequencing and molecular cloning, we showed that Csyc is a potential candidate gene, which encodes a yellow stripe-like (YSL) transporter. The T to C mutation in the promoter region of Csyc caused the yellow cotyledon phenotype in yc412. Compared to YZU027A (WT), the expression of Csyc was significantly downregulated in the cotyledons of yc412. Silencing of Csyc in cucumber via virus-induced gene silencing resulted in chlorotic leaves, mainly by suppressing the chlorophyll content. Furthermore, a comparative transcriptome analysis revealed that chloroplast-related genes and chlorophyll biosynthesis genes were significantly downregulated in yc412 cotyledons. Our results provide new insights into the molecular function of the YSL transporter in plant chloroplast development and chlorophyll synthesis.

The transcription factor WRKY12 negatively regulates iron entry into seeds in Arabidopsis.

Yellow Stripe 1-Like 1 (YSL1) and Yellow Stripe 1-Like 3 (YSL3) transport metal-nicotianamine (NA) complexes to leaves, pollen, and developing seeds and play an important role in regulating iron (Fe) accumulation during the seed development and maturation stages; however, how their gene transcript levels are regulated remains unknown. In this study, we used yeast one-hybrid screening to identify a transcription factor, WRKY12, in Arabidopsis that directly regulates the transcription levels of YSL1 and YSL3 genes. WRKY12 has opposite expression patterns to YSL1 and YSL3. wrky12 mutants are tolerant to Fe deficiency, whereas WRKY12 overexpression lines are sensitive to Fe deficiency. During the development and maturation of seeds, WRKY12 can directly bind to the promoters of YSL1 and YSL3 and inhibit their expression. Genetic analysis showed that WRKY12 functions upstream of YSL1 and YSL3 in Fe intake during the seed development and maturation stages. Together, our results suggest that WRKY12 negatively regulates the iron intake in plant seeds by inhibiting the expression of YSL1 and YSL3.

Advances in Genes-Encoding Transporters for Cadmium Uptake, Translocation, and Accumulation in Plants.

Cadmium (Cd) is a heavy metal that is highly toxic for plants, animals, and human beings. A better understanding of the mechanisms involved in Cd accumulation in plants is beneficial for developing strategies for either the remediation of Cd-polluted soils using hyperaccumulator plants or preventing excess Cd accumulation in the edible parts of crops and vegetables. As a ubiquitous heavy metal, the transport of Cd in plant cells is suggested to be mediated by transporters for essential elements such as Ca, Zn, K, and Mn. Identification of the genes encoding Cd transporters is important for understanding the mechanisms underlying Cd uptake, translocation, and accumulation in either crop or hyperaccumulator plants. Recent studies have shown that the transporters that mediate the uptake, transport, and accumulation of Cd in plants mainly include members of the natural resistance-associated macrophage protein (Nramp), heavy metal-transporting ATPase (HMA), zinc and iron regulated transporter protein (ZIP), ATP-binding cassette (ABC), and yellow stripe-like (YSL) families. Here, we review the latest advances in the research of these Cd transporters and lay the foundation for a systematic understanding underlying the molecular mechanisms of Cd uptake, transport, and accumulation in plants.

Genome-wide understanding of evolutionary and functional relationships of rice Yellow Stripe-Like (YSL) transporter family in comparison with other plant species

Genome-wide understanding of evolutionary and functional relationships of rice Yellow Stripe-Like (YSL) transporter family in comparison with other plant species

Efficiency of transporter genes and proteins in hyperaccumulator plants for metals tolerance in wastewater treatment: Sustainable technique for metal detoxification

Environmental contamination of heavy metals is now becoming increasingly a concern and a significant problem due to its harmful effects worldwide. The plant-meditated approach is encouraging to eliminate toxins avoiding side effects from polluted wastewater. For the development of appropriate plant species for the mechanisms of heavy metal absorption, transport, detoxification, identification, and signaling pathways would be important facts. Transporter genes like ATP-phosphoribosyl transferase (ATP-PRT), Yellow Stripe-like (YSL), NAS (nicotinamide synthase), SAMS (S-adenosyl-methionine synthetase), FER (ferritin Fe (III) binding), HMA (heavy metal ATPase), IREG (iron-regulated transporter), and proteins like cation diffusion facilitators (CDF), ZRT, IRT-like protein (ZIP), and natural resistance-associated macrophage protein (NRAMP) are active in heavy metal accumulation, translocalisation, sequestration, and resistance. Besides, chelating agents and metabolites can be used either to increase heavy metal bioavailability, which facilitates heavy metal accumulation in plants and further promote plant growth and fitness. This review paper addresses key roles and potential transporter genes and proteins for the remediation of heavy metals from hyperaccumulator plants. This review specifically focuses on the efficacy of transporter genes and proteins in hyperaccumulator plants in metal restoration, discussing the use of these plants for wastewater treatment processes. • Discusses the efficiency of hyperaccumulator plant for removal of metals. • Summarizes transporter gene and proteins in heavy metal uptake and tolerance. • Proposes improved prospects for wastewater treatment.

Shrunken4 is a mutant allele of ZmYSL2 that affects aleurone development and starch synthesis in maize.

Minerals are stored in the aleurone layer and embryo during maize seed development, but how they affect endosperm development and activity is unclear. Here, we cloned the gene underlying the classic maize kernel mutant shrunken4 (sh4) and found that it encodes the YELLOW STRIPE-LIKE oligopeptide metal transporter ZmYSL2. sh4 kernels had a shrunken phenotype with developmental defects in the aleurone layer and starchy endosperm cells. ZmYSL2 showed iron and zinc transporter activity in Xenopus laevis oocytes. Analysis using a specific antibody indicated that ZmYSL2 predominately accumulated in the aleurone and sub-aleurone layers in endosperm and the scutellum in embryos. Specific iron deposition was observed in the aleurone layer in wild-type kernels. In sh4, however, the outermost monolayer of endosperm cells failed to accumulate iron and lost aleurone cell characteristics, indicating that proper functioning of ZmYSL2 and iron accumulation are essential for aleurone cell development. Transcriptome analysis of sh4 endosperm revealed that loss of ZmYSL2 function affects the expression of genes involved in starch synthesis and degradation processes, which is consistent with the delayed development and premature degradation of starch grains in sh4 kernels. Therefore, ZmYSL2 is critical for aleurone cell development and starchy endosperm cell activity during maize seed development.

Medicago truncatula Yellow Stripe‐Like7 encodes a peptide transporter participating in symbiotic nitrogen fixation

Yellow Stripe-Like (YSL) proteins are a family of plant transporters that are typically involved in transition metal homeostasis. Three of the four YSL clades (I, II and IV) transport metals complexed with the non-proteinogenic amino acid nicotianamine or its derivatives. No such capability has been shown for any member of clade III, but the link between these YSLs and metal homeostasis could be masked by functional redundancy. We studied the role of the clade III YSL protein MtSYL7 in Medicago truncatula nodules. MtYSL7, which encodes a plasma membrane-bound protein, is mainly expressed in the pericycle and cortex cells of the root nodules. Yeast complementation assays revealed that MtSYL7 can transport short peptides. M. truncatula transposon insertion mutants with decreased expression of MtYSL7 had lower nitrogen fixation rates and showed reduced plant growth whether grown in symbiosis with rhizobia or not. YSL7 mutants accumulated more copper and iron in the nodules, which is likely to result from the increased expression of iron uptake and delivery genes in roots. Taken together, these data suggest that MtYSL7 plays an important role in the transition metal homeostasis of nodules and symbiotic nitrogen fixation.

TaYS1A, a Yellow Stripe-Like Transporter Gene, Is Required for Wheat Resistance to Puccinia striiformis f. sp. Tritici.

Yellow stripe-like (YSL) transporters are required for the transportation of metal-phytosiderophores and are structurally related to metal-nicotianamine complexes. Some studies also reported the involvement of YSL transporters in pathogen-induced defense. However, the molecular mechanisms of YSL genes involved in biotic stress responses are still not clear, especially in cereal crops. This study aimed to functionally characterize TaYS1A during the interaction of wheat and Puccinia striiformis f. sp. tritici (Pst), the causal agent of stripe rust disease. TaYS1A was localized in the cell membrane of wheat protoplasts and Nicotiana benthamiana cells. TaYS1A was significantly up-regulated in wheat leaves after being infected with the avirulent Pst isolate CYR23 and after treatment with salicylic acid (SA). Silencing of TaYS1A by the virus-induced gene silencing method enhanced the susceptibility of wheat to Pst accompanied by reducing the accumulation of SA and H2O2 and down-regulating the transcriptions of TaPR1 and TaPR2. In addition, TaYS1A was found to interact with TaNH2, a homolog of OsNH2, by yeast-two-hybrid assay, and silencing of TaYS1A diminished the expression of TaNH2. Our findings suggested the existence of positive regulation of TaYS1A in providing resistance against Pst by modulating SA-induced signaling and offered new insight into the biological role of YSL in wheat against pathogens.

The Medicago truncatula Yellow Stripe1-Like3 gene is involved in vascular delivery of transition metals to root nodules.

Symbiotic nitrogen fixation carried out in legume root nodules requires transition metals. These nutrients are delivered by the host plant to the endosymbiotic nitrogen-fixing bacteria living within the nodule cells, a process in which vascular transport is essential. As members of the Yellow Stripe-Like (YSL) family of metal transporters are involved in root to shoot transport, they should also be required for root to nodule metal delivery. The genome of the model legume Medicago truncatula encodes eight YSL proteins, four of them with a high degree of similarity to Arabidopsis thaliana YSLs involved in long-distance metal trafficking. Among them, MtYSL3 is a plasma membrane protein expressed by vascular cells in roots and nodules and by cortical nodule cells. Reducing the expression level of this gene had no major effect on plant physiology when assimilable nitrogen was provided in the nutrient solution. However, nodule functioning was severely impaired, with a significant reduction of nitrogen fixation capabilities. Further, iron and zinc accumulation and distribution changed. Iron was retained in the apical region of the nodule, while zinc became strongly accumulated in the nodule veins in the ysl3 mutant. These data suggest a role for MtYSL3 in vascular delivery of iron and zinc to symbiotic nitrogen fixation.

Melatonin confers cadmium tolerance by modulating critical heavy metal chelators and transporters in radish plants.

Cadmium (Cd) is an environmental pollutant that causes health hazard to living organisms. Melatonin (MT) has emerged as a ubiquitous pleiotropic molecule capable of coordinating heavy metal (HM) stresses in plants. However, it remains unclear how melatonin mediates Cd homeostasis and detoxification at transcriptional and/or post-transcriptional levels in radish. Herein, the activities of five key antioxidant enzymes were increased, while root and shoot Cd contents were dramatically decreased by melatonin. A combined small RNA and transcriptome sequencing analysis showed that 14 differentially expressed microRNAs (DEMs) and 966 differentially expressed genes (DEGs) were shared between the Cd and Cd+MT conditions. In all, 23 and ten correlated miRNA-DEG pairs were identified in Con vs. Cd and Con vs. Cd+MT comparisons, respectively. Several DEGs encoding yellow stripe 1-like (YSL), heavy metal ATPases (HMA), and ATP-binding cassette (ABC) transporters were involved in Cd transportation and sequestration in radish. Root exposure to Cd2+ induced several specific signaling molecules, which consequently trigger some HM chelators, transporters, and antioxidants to achieve reactive oxygen species (ROS) scavenging and detoxification and eliminate Cd toxicity in radish plants. Notably, transgenic analysis revealed that overexpression of the RsMT1 (Metallothionein 1) gene could enhance Cd tolerance of tobacco plants, indicating that the exogenous melatonin confers Cd tolerance, which might be attributable to melatonin-mediated upregulation of RsMT1 gene in radish plants. These results could contribute to dissecting the molecular basis governing melatonin-mediated Cd stress response in plants and pave the way for high-efficient genetically engineering low-Cd-content cultivars in radish breeding programs.

Molecular Analysis of Iron Deficiency Response in Hexaploid Wheat

Iron deficiency leads to severe chlorosis in crop plants, including wheat, thereby reducing total yield and quality. Furthermore, grains of most bread wheat varieties are poor source of iron, which is vital for human nutrition. Despite the significance, iron uptake and translocation mechanisms in bread wheat have not been studied in detail, particularly under iron limited growth conditions. In this study, bread wheat plants were grown under iron deficiency stress until maturity. Data were collected at three distinct developmental time points during grain-filling. The plants experiencing low iron availability exhibited significantly lower chlorophyll content as well as low iron concentration in leaves and grains. The expression levels of bread wheat genes homologous to iron deficiency responsive genes of rice, barley, and Arabidopsis were significantly changed under iron deficiency stress. The wheat homologs of genes involved in phytosiderophore (PS) synthesis and transport were significantly up-regulated in the iron-deficient roots through all development stages, confirming an important role of deoxymugineic acid (DMA) in iron acquisition. The up-regulation of NICOTIANAMINE SYNTHASE (NAS) and DEOXYMUGINEIC ACID SYNTHASE (DMAS) in flag leaves and grains suggested the involvement of nicotianamine (NA) and DMA in iron chelation and translocation in wheat, particularly at the commencement of grain-filling. In line with this, the homolog of gene encoding TRANSPORTER OF MUGINEIC ACID (TOM) was up-regulated in the wheat roots under iron deficiency. Additionally, genes encoding long-distance iron transporter YELLOW STRIPE-LIKE (YSL), the vacuolar transporter NATURAL RESISTANCE ASSOCIATED MACROPHAGE PROTEIN (NRAMP), and the transcription factor BASIC HELIX-LOOP-HELIX (bHLH), were also up-regulated upon iron starvation. A tissue specific and growth stage specific gene expression differences in response to iron deficiency stress were observed, providing new insights into iron translocation, storage and regulation in bread wheat.

The search for candidate genes associated with natural variation of grain Zn accumulation in barley.

Combating hidden hunger through molecular breeding of nutritionally enriched crops requires a better understanding of micronutrient accumulation. We studied natural variation in grain micronutrient accumulation in barley (Hordeum vulgare L.) and searched for candidate genes by assessing marker-trait associations (MTAs) and by analyzing transcriptional differences between low and high zinc (Zn) accumulating cultivars during grain filling. A collection of 180 barley lines was grown in three different environments. Our results show a pronounced variation in Zn accumulation, which was under strong genotype influence across different environments. Genome-wide association mapping revealed 13 shared MTAs. Across three environments, the most significantly associated marker was on chromosome 2H at 82.8 cM and in close vicinity to two yellow stripe like (YSL) genes. A subset of two pairs of lines with contrasting Zn accumulation was chosen for detailed analysis. Whole ears and flag leaves were analyzed 15 days after pollination to detect transcriptional differences associated with elevated Zn concentrations in the grain. A putative α-amylase/trypsin inhibitor CMb precursor was decidedly higher expressed in high Zn cultivars in whole ears in all comparisons. Additionally, a gene similar to barley metal tolerance protein 5 (MTP5) was found to be a potential candidate gene.

Gene expression and biochemical response of giant reed under Ni and Cu stress

Giant reed (Arundo donax) has proved to be effective in detoxification, accumulation and tolerance of toxic metals. The present study explored the stress response of giant reed against Cu and Ni stress. The effect of metal stress was studied on dry weight, chlorophyll pigments antioxidant enzymes production and selected genes expression. The accumulation of heavy metals increased in a concentration-dependent manner and depicted toxicity symptoms in leaves beyond 75 mg/L of Cu or Ni. Oxidative stress was evident in giant reed under highest exposure of Ni and Cu which increased antioxidants activities (SOD, POD and CAT). It was observed that metal transport and detoxification were possible due to the expression of glutathione reductase, Natural Resistance-Associated Macrophage Protein (NRAMP) and Yellow Stripe-Like (YSL) genes. These insights into the genetic basis of a successful remediating plant species will be useful in understanding heavy metals tolerance in giant reed.

The Yellow Stripe-Like (YSL) Gene Functions in Internal Copper Transport in Peanut

Copper (Cu) is involved in fundamental biological processes for plant growth and development. However, Cu excess is harmful to plants. Thus, Cu in plant tissues must be tightly regulated. In this study, we found that the peanut Yellow Stripe-Like family gene AhYSL3.1 is involved in Cu transport. Among five AhYSL genes, AhYSL3.1 and AhYSL3.2 were upregulated by Cu deficiency in peanut roots and expressed mainly in young leaves. A yeast complementation assay suggested that the plasma membrane-localized AhYSL3.1 was a Cu-nicotianamine complex transporter. High expression of AhYSL3.1 in tobacco and rice plants with excess Cu resulted in a low concentration of Cu in young leaves. These transgenic plants were resistant to excess Cu. The above results suggest that AhYSL3.1 is responsible for the internal transport of Cu in peanut.

OsYSL16 is Required for Preferential Cu Distribution to Floral Organs in Rice.

Deficiency of copper (Cu) causes low fertility in many plant species, but the molecular mechanisms underlying distribution of Cu to the floral organs are poorly understood. Here, we found that a member of yellow-stripe like (YSL) family, YSL16 encoding the Cu-nicotianamine (Cu-NA) transporter, was highly expressed in the rachilla, with less expression in the palea and lemma of rice (Oryza sativa). β-Glucuronidase (GUS) staining of transgenic rice carrying the OsYSL16 promoter-GUS showed that OsYSL16 was mainly expressed in vascular bundles of the rachilla as well as the palea and lemma. Knockout of OsYSL16 resulted in decreased Cu distribution to the stamens, but increased distribution to the palea and lemma. A short-term (24 h) 65Cu labeling experiment confirmed increased Cu concentration of palea and lemma in the mutant. Furthermore, we found that redistribution of Cu from the palea and lemma was impaired in the osysl16 mutant after exposure to Cu-free solution. The osysl16 mutant showed low pollen germination, but this was rescued by addition of Cu in the medium. Our results indicate that OsYSL16 expressed in the vascular bundles of the rachilla is important for preferential distribution of Cu to the stamens, while OsYSL16 in vascular bundles of the palea and lemma is involved in Cu redistribution under Cu-limited conditions in rice.

Genome-wide analysis indicates diverse physiological roles of the turnip (Brassica rapa var. rapa) oligopeptide transporters gene family

Oligopeptide transporters (OPTs) encode integral membrane-localized proteins and have a broad range of substrate transport capabilities. Here, 28 BrrOPT genes were identified in the turnip. Phylogenetic analyses revealed two well-supported clades in the OPT family, containing 15 BrrOPTs and 13 BrrYSLs. The exon/intron structure of OPT clade are conserved but the yellow stripe-like (YSL) clade was different. The exon/intron of the YSL clade possesses structural differences, whereas the YSL class motifs structure are conserved. The OPT genes are distributed unevenly among the chromosomes of the turnip genome. Phylogenetic and chromosomal distribution analyses revealed that the expansion of the OPT gene family is mainly attributable to segmental duplication. For the expression profiles at different developmental stages, a comprehensive analysis provided insights into the possible functional divergence among members of the paralog OPT gene family. Different expression levels under a variety of ion deficiencies also indicated that the OPT family underwent functional divergence during long-term evolution. Furthermore, BrrOPT8.1, BrrYSL1.2, BrrYSL1.3, BrrYSL6 and BrrYSL9 responded to Fe(II) treatments and BrrYSL7 responded to calcium treatments, BrrYSL6 responded to multiple treatments in root, suggesting that turnip OPTs may be involved in mediating cross-talk among different ion deficiencies. Our data provide important information for further functional dissection of BrrOPTs, especially in transporting metal ions and nutrient deficiency stress adaptation.

Overexpression of a Miscanthus sacchariflorus yellow stripe-like transporter MsYSL1 enhances resistance of Arabidopsis to cadmium by mediating metal ion reallocation

The yellow stripe-like (YSL) family of transporters mediates the uptake, translocation, and distribution of various mineral elements in vivo by transferring metal ions chelated with phytosiderophore or nicotianamine (NA). However, little is known about the roles of the YSL genes against cadmium in planta. In this study, we first cloned and characterized a vital member of the YSL gene family, MsYSL1, from the bioenergy plant Miscanthus sacchariflorus. MsYSL1 localized in the plasma membrane and was widely expressed throughout the whole seedling with the highest expression level in the stem. In addition, its expression in the root was stimulated by excess manganese (Mn), cadmium (Cd), and lead, and a shortage of iron (Fe), zinc (Zn), and copper. Functional complementation in yeast indicated that MsYSL1 showed transport activity for Fe(II)–NA and Zn–NA, but not for Cd–NA. Although they exhibited no significant differences versus the wild type under normal cultivation conditions, MsYSL1-overexpressing Arabidopsis lines displayed a higher resistance to Cd accompanied by longer root lengths, lower Cd, Zn, and Mn levels in roots, and higher Cd, Fe, and Mn translocation ratios under Cd stress. Moreover, genes related to NA synthesis, metal translocation, long-distance transport, and Cd exclusion were highly induced in transgenic lines under Cd stress. Thus, MsYSL1 may be an essential transporter for diverse metal–NAs to participate in the Cd detoxification by mediating the reallocation of other metal ions.

The iron-chelate transporter OsYSL9 plays a role in iron distribution in developing rice grains.

Rice OsYSL9 is a novel transporter for Fe(II)-nicotianamine and Fe(III)-deoxymugineic acid that is responsible for internal iron transport, especially from endosperm to embryo in developing seeds. Metal chelators are essential for safe and efficient metal translocation in plants. Graminaceous plants utilize specific ferric iron chelators, mugineic acid family phytosiderophores, to take up sparingly soluble iron from the soil. Yellow Stripe 1-Like (YSL) family transporters are responsible for transport of metal-phytosiderophores and structurally similar metal-nicotianamine complexes. Among the rice YSL family members (OsYSL) whose functions have not yet been clarified, OsYSL9 belongs to an uncharacterized subgroup containing highly conserved homologs in graminaceous species. In the present report, we showed that OsYSL9 localizes mainly to the plasma membrane and transports both iron(II)-nicotianamine and iron(III)-deoxymugineic acid into the cell. Expression of OsYSL9 was induced in the roots but repressed in the nonjuvenile leaves in response to iron deficiency. In iron-deficient roots, OsYSL9 was induced in the vascular cylinder but not in epidermal cells. Although OsYSL9-knockdown plants did not show a growth defect under iron-sufficient conditions, these plants were more sensitive to iron deficiency in the nonjuvenile stage compared with non-transgenic plants. At the grain-filling stage, OsYSL9 expression was strongly and transiently induced in the scutellum of the embryo and in endosperm cells surrounding the embryo. The iron concentration was decreased in embryos of OsYSL9-knockdown plants but was increased in residual parts of brown seeds. These results suggested that OsYSL9 is involved in iron translocation within plant parts and particularly iron translocation from endosperm to embryo in developing seeds.

NaCl alleviates iron deficiency through facilitating root cell wall iron reutilization and its translocation to the shoot in Arabidopsis thaliana

The present study was undertaken to investigate the mechanism underlying NaCl-alleviated Fe (iron) deficiency in Arabidopsis thaliana Col-0 (Columbia ecotype). Six-week-old Col-0 with rosettes of similar diameters was grown in full-strength nutrient solution lacking Fe (Fe-deficient) or full-strength nutrient solution (Fe-sufficient) with or without 10 mM NaCl for 7 days. Roots were then harvested for analysis of total Fe content, soluble Fe concentration, and abscisic acid (ABA) content as well as for cell wall and RNA extraction, while shoots were harvested for total Fe content and soluble Fe concentration measurements. The total Fe content and the soluble Fe concentration were higher in roots and shoots of −Fe + NaCl-treated plants than −Fe-treated plants, whereas Fe retention in the cell wall was reduced, suggesting the presence of a cell wall Fe-reutilization mechanism. This conclusion was confirmed by the observation that less hemicelluloses Fe was found in plants under −Fe + NaCl treatment. In addition, associated to the upregulation of genes related to the long-distance transport of Fe, such as FRD3 (FERRIC REDUCTASE DEFECTIVE3), YSL2 (YELLOW STRIPE-LIKE), and NAS1 (NICOTIANAMINE SYNTHASE1) under −Fe + NaCl treatment, more Fe was available in shoots. Furthermore, endogenous ABA is involved in NaCl-alleviated Fe deficiency, as the addition of Flu (fluridone), an inhibitor of ABA biosynthesis, abolished the positive effect of NaCl on Fe deficiency. Under −Fe condition, NaCl not only is involved in the reutilization of cell wall Fe but also participates in the translocation of Fe from root to shoot in Arabidopsis, partially through its effect on ABA contents.