Reduced expression of bZIP19 and bZIP23 increases zinc and cadmium accumulation in Arabidopsis halleri.

Zinc is an essential micronutrient for all living organisms. When challenged by zinc-limiting conditions, Arabidopsis thaliana plants use a strategy centered on two transcription factors, bZIP19 and bZIP23, to enhance the expression of several zinc transporters to improve their zinc uptake capacity. In the zinc and cadmium hyperaccumulator plant Arabidopsis halleri, highly efficient root-to-shoot zinc translocation results in constitutive local zinc deficiency in roots and in constitutive high expression of zinc deficiency-responsive ZIP genes, supposedly boosting zinc uptake and accumulation. Here, to disrupt this process and to analyze the functions of AhbZIP19, AhbZIP23 and their target genes in hyperaccumulation, the genes encoding both transcriptional factors were knocked down using artificial microRNAs (amiRNA). Although AhbZIP19, AhbZIP23, and their ZIP target genes were downregulated, amiRNA lines surprisingly accumulated more zinc and cadmium compared to control lines in both roots and shoot driving to shoot toxicity symptoms. These observations suggested the existence of a substitute metal uptake machinery in A. halleri to maintain hyperaccumulation. We propose that the iron uptake transporter AhIRT1 participates in this alternative pathway in A. halleri.

Tattoo inks: evaluation of cellular responses and analysis of some trace metals.

After tattoo application, inks remain in the skin, mostly in the dermal layer, and manufacturers use inks that have not been adequately evaluated for safety in tattoo production. In this study, the metal contents (Cd, Hg, Pb, and Cr) of tattoo inks available in the Turkish market were determined and the relationship between cell viability and inflammatory response of the detected metal levels was investigated. Nine tattoo inks (3 colors) from 3 different brands abbreviated as E, I, and W were examined. ICP-MS was used for element analysis. The viability of human keratinocyte cells was determined by the WST-1 assay following ink exposures at various dilutions. IL-18 levels were measured in cell culture supernatant by ELISA method following ink or metal (Cd, Cr, Hg, and Pb) exposures. The concentrations of trace elements were found in inks as follows: Cd, 0.0641-1.3857; Hg, 0.0204-0.2675; Pb, 0.8527-6.5981; Cr, 0.1731-45.3962 µg mL-1. It was observed that the levels of Pb and especially Cr in the samples exceeded the limit values. Tattoo inks reduced the cell viability in a dose- and color-dependent manner. IL-18 release was significantly increased in all groups except Cr and black ink of brand I treated cells (p < 0.05). Our results show that the metal contents of tattoo inks exceed Council of Europe Resolution values in some samples and some inks induce immune system activation (IL-18 secretion) and cytotoxic effects. It is thought that these findings may contribute to the toxic/adverse effects of tattoo inks commonly used.

Transcriptomics, proteomics, and metabolomics interventions prompt crop improvement against metal(loid) toxicity.

The escalating challenges posed by metal(loid) toxicity in agricultural ecosystems, exacerbated by rapid climate change and anthropogenic pressures, demand urgent attention. Soil contamination is a critical issue because it significantly impacts crop productivity. The widespread threat of metal(loid) toxicity can jeopardize global food security due to contaminated food supplies and pose environmental risks, contributing to soil and water pollution and thus impacting the whole ecosystem. In this context, plants have evolved complex mechanisms to combat metal(loid) stress. Amid the array of innovative approaches, omics, notably transcriptomics, proteomics, and metabolomics, have emerged as transformative tools, shedding light on the genes, proteins, and key metabolites involved in metal(loid) stress responses and tolerance mechanisms. These identified candidates hold promise for developing high-yielding crops with desirable agronomic traits. Computational biology tools like bioinformatics, biological databases, and analytical pipelines support these omics approaches by harnessing diverse information and facilitating the mapping of genotype-to-phenotype relationships under stress conditions. This review explores: (1) the multifaceted strategies that plants use to adapt to metal(loid) toxicity in their environment; (2) the latest findings in metal(loid)-mediated transcriptomics, proteomics, and metabolomics studies across various plant species; (3) the integration of omics data with artificial intelligence and high-throughput phenotyping; (4) the latest bioinformatics databases, tools and pipelines for single and/or multi-omics data integration; (5) the latest insights into stress adaptations and tolerance mechanisms for future outlooks; and (6) the capacity of omics advances for creating sustainable and resilient crop plants that can thrive in metal(loid)-contaminated environments.

Metal Toxicity: Effects on Energy Metabolism in Fish.

Metals are dispersed in natural environments, particularly in the aquatic environment, and accumulate, causing adverse effects on aquatic life. Moreover, chronic polymetallic water pollution is a common problem, and the biological effects of exposure to complex mixtures of metals are the most difficult to interpret. In this review, metal toxicity is examined with a focus on its impact on energy metabolism. Mechanisms regulating adenosine triphosphate (ATP) production and reactive oxygen species (ROS) emission are considered in their dual roles in the development of cytotoxicity and cytoprotection, and mitochondria may become target organelles of metal toxicity when the transmembrane potential is reduced below its phosphorylation level. One of the main consequences of metal toxicity is additional energy costs, and the metabolic load can lead to the disruption of oxidative metabolism and enhanced anaerobiosis.

Monitoring of university wastewater within the sewage system and its performance evaluation through integrated constructed wetlands.

Rapid increase in population and industrialization has not only improved the lifestyle but adversely affected the quality and availability of water leading to ample amount of wastewater generation. The major contribution towards wastewater production is from sewage. Regular monitoring and treatment of sewage water is necessary to conserve and enhance the quality of water. The present study focuses on monitoring of sewage water within the sewage system of a residential university. A total of 16 samples from different manholes were collected for physicochemical and heavy metals analysis and compared with final effluent collected from integrated constructed wetlands (ICWs) to assess its removal efficiency. The mean concentrations of influent and effluent were compared with national environmental quality standards (NEQS) for municipal discharge (pH 6-9, COD 150 mg/L, TSS 200 mg/L and TDS 3500 mg/L) and international agricultural reuse standards (IARS) (pH 6-8, COD <150 mg/L, TSS < 100 mg/L) respectively. Among all physicochemical parameters, influent values for chemical oxygen demand (COD) (169.56-258.36) mg/L exceeded the limit of NEQS for discharge into inland waters, whereas for total suspended solids (TSS) the concentration exceeded for discharge into STP (406 mg/L) and inland waters (202.33 mg/L). However, effluent concentrations for all the parameters were found within the permissible limit set by IARS. The removal efficiency for different parameters such as phosphate- phosphorus (PO43-P), COD, TSS, total dissolved solids (TDS) and total kjeldahl nitrogen (TKN) were 52, 53, 54, 35, and 36%, respectively. Heavy metal concentrations were compared with WHO guidelines among which lead (Pb) in effluent and chromium (Cr) in influent exceeded the limit (Pb 0.01 and Cr 0.05 mg/L). Interpolation results showed that zone 2 was highly contaminated in comparison to zone 1 & 3. Statistical analysis showed that correlation of physicochemical parameters and heavy metals was found significant (p < 0.05).

Molecular Regulation and Evolution of Cytokinin Signaling in Plant Abiotic Stresses.

The sustainable production of crops faces increasing challenges from global climate change and human activities, which leads to increasing instances of many abiotic stressors to plants. Among the abiotic stressors, drought, salinity and excessive levels of toxic metals cause reductions in global agricultural productivity and serious health risks for humans. Cytokinins (CKs) are key phytohormones functioning in both normal development and stress responses in plants. Here, we summarize the molecular mechanisms on the biosynthesis, metabolism, transport and signaling transduction pathways of CKs. CKs act as negative regulators of both root system architecture plasticity and root sodium exclusion in response to salt stress. The functions of CKs in mineral-toxicity tolerance and their detoxification in plants are reviewed. Comparative genomic analyses were performed to trace the origin, evolution and diversification of the critical regulatory networks linking CK signaling and abiotic stress. We found that the production of CKs and their derivatives, pathways of signal transduction and drought-response root growth regulation are evolutionarily conserved in land plants. In addition, the mechanisms of CK-mediated sodium exclusion under salt stress are suggested for further investigations. In summary, we propose that the manipulation of CK levels and their signaling pathways is important for plant abiotic stress and is, therefore, a potential strategy for meeting the increasing demand for global food production under changing climatic conditions.

Endophytic yeast protect plants against metal toxicity by inhibiting plant metal uptake through an ethylene-dependent mechanism.

Toxic metal pollution requires significant adjustments in plant metabolism. Here, we show that the plant microbiota plays an important role in this process. The endophytic Sporobolomyces ruberrimus isolated from a serpentine population of Arabidopsis arenosa protected plants against excess metals. Coculture with its native host and Arabidopsis thaliana inhibited Fe and Ni uptake. It had no effect on host Zn and Cd uptake. Fe uptake inhibition was confirmed in wheat and rape. Our investigations show that, for the metal inhibitory effect, the interference of microorganisms in plant ethylene homeostasis is necessary. Application of an ethylene synthesis inhibitor, as well as loss-of-function mutations in canonical ethylene signalling genes, prevented metal uptake inhibition by the fungus. Coculture with S. ruberrimus significantly changed the expression of Fe homeostasis genes: IRT1, OPT3, OPT6, bHLH38 and bHLH39 in wild-type (WT) A. thaliana. The expression pattern of these genes in WT plants and in the ethylene signalling defective mutants significantly differed and coincided with the plant accumulation phenotype. Most notably, down-regulation of the expression of IRT1 solely in WT was necessary for the inhibition of metal uptake in plants. This study shows that microorganisms optimize plant Fe and Ni uptake by fine-tuning plant metal homeostasis.

Regulatory mechanisms of sulfur metabolism affecting tolerance and accumulation of toxic trace metals and metalloids in plants.

Soil contamination with trace metals and metalloids can cause toxicity to plants and threaten food safety and human health. Plants have evolved sophisticated mechanisms to cope with excess trace metals and metalloids in soils, including chelation and vacuolar sequestration. Sulfur-containing compounds, such as glutathione and phytochelatins, play a crucial role in their detoxification, and sulfur uptake and assimilation are regulated in response to the stress of toxic trace metals and metalloids. This review focuses on the multi-level connections between sulfur homeostasis in plants and responses to such stresses, especially those imposed by arsenic and cadmium. We consider recent progress in understanding the regulation of biosynthesis of glutathione and phytochelatins and of the sensing mechanism of sulfur homeostasis for tolerance of trace metals and metalloids in plants. We also discuss the roles of glutathione and phytochelatins in controlling the accumulation and distribution of arsenic and cadmium in plants, and possible strategies for manipulating sulfur metabolism to limit their accumulation in food crops.

Silicon modification improves biochar's ability to mitigate cadmium toxicity in tomato by enhancing root colonization of plant-beneficial bacteria.

Modification of biochar, such as impregnation with minerals, can improve biochar's efficacy to mitigate heavy metal toxicity in plants. Biochar amendments can alter plant rhizosphere microbiome, which has profound effects on plant growth and fitness. Here, we tested whether rhizosphere microbiome is involved in the ability of silicon (Si)-modified biochar to mitigate cadmium toxicity in tomato (Solanum lycopersicum L.). We demonstrated that Si modification altered biochar's physico-chemical properties and enhanced its ability to mitigate cadmium toxicity in tomato. Particularly, the Si-modified biochar contained higher content of Si and increased plant-available Si content in the soil. The rhizosphere microbiome transplant experiment showed that changes in rhizosphere microbiome contributed to the mitigation of cadmium toxicity by biochar amendments. The raw biochar and Si-modified biochar differently altered tomato rhizosphere bacterial community composition. Both biochars, especially the Si-modified biochar, promoted specific bacterial taxa (e.g., Sphingomonas, Lysobacter and Pseudomonas spp.). Subsequent culturing found these promoted bacteria could mitigate cadmium toxicity in tomato. Moreover, both biochars stimulated tomato to recruit plant-beneficial bacteria with Si-modified biochar having stronger stimulatory effects, indicating that the positive effects of biochar on plant-beneficial bacteria was partially mediated via the host plant. Overall, Si modification enhanced biochar's ability to mitigate cadmium toxicity, which was linked to the stimulatory effects on plant-beneficial bacteria.

Moringa oleifera smoke induced positive changes in biochemical, metabolic, and antioxidant profile of rice seedling under cadmium stress.

Cadmium as a heavy metal contaminates the agricultural soil and effect plant growth due to rapid increases in industrialization and anthropogenic activities. Smoke water of Moringa oleifera was used in the current study to alleviate the effect of cadmium on the physiological, biochemical, metabolic, and antioxidant profile of Basmati 385 and Shaheen Basmati seedling. Cadmium stress of 100, 200, and 400 µM were given to 28 days-old seedlings along with smoke water (1:1,000) for one week in hydroponic culture. As a result, Cd+2 toxicity negatively affects the seedling length, fresh and dry weight, photosynthetic pigment, and electrolytes leakage, while the application of smoke water alleviated those effects. Furthermore, Cd+2 content, cell injury, metabolic parameters (proline, total soluble sugar), and antioxidants (peroxidase, catalase) were increased with increasing Cd+2 concentration while smoke water-treated seedlings showed reduction at high concentration. From present study, it can be concluded that smoke water had some regulatory compound which could reduce the Cd+2 stress level in rice seedlings and improve plant growth.

Moringa (Moringa oleifera) is a famous medicinal plant. Its fruits, roots, leaves, and flowers are used as vegetables in different part of the world. Moringa leaves are rich source of vitamin A, C riboflavin, beta carotenoid, iron, and phenolic acid and also reported for antioxidant properties. The unique aspect of current study is use to M. oliferia leaves for the preparation of smoke water, because of its nutritional and antioxidant properties and further its effects was observed on rice seedling under cadmium stress, which has not been evaluated or reported earlier.

Vitamin K2 protects against aluminium chloride-mediated neurodegeneration.

Recent studies have shown that, coupled with other environmental factors, aluminium exposure may lead to neurodegeneration resulting in cognitive impairment resembling Alzheimer's disease. Menaquinone, a form of vitamin K2, aids in maintaining healthy bones and avoids coronary calcification. It also has anti-inflammatory and antioxidant properties. Here, we study the neuroprotective effects of vitamin K2 (MK-7) using the animal model of Alzheimer's disease (AD). Aluminium chloride (AlCl3; 100 mg/kg for 3 weeks orally) was administered to Swiss albino mice to induce neurodegeneration and Vitamin K2 (100 g/kg for 3 weeks orally) was applied as treatment. This was followed by behavioural studies to determine memory changes. The behavioural observations correlated with proinflammatory, oxidative, and brain histopathological changes in AlCl3-treated animals with or without vitamin K2 treatment. AlCl3 administration led to memory decline which was partially restored in Vitamin K2 treated animals. Myeloperoxidase levels in the brain increased due to AlCl3-mediated inflammation, which Vitamin K2 prevented. The acetylcholine esterase and oxidative stress markers induced by AlCl3 were reversed by Vitamin K2. Also, Vitamin K2 helps to restore hippocampal BDNF levels and reduced the amyloid ß accumulation in AlCl3-administered animals. Additionally, Vitamin K2 protected the hippocampal neurons against AlCl3-mediated damage as observed in histopathological studies. We conclude that Vitamin K2 could partially reverse AlCl3-mediated cognitive decline. It increases acetylcholine and BDNF levels while reducing oxidative stress, neuroinflammation, and ß-amyloid deposition, thus protecting the hippocampal neurons from AlCl3-mediated damage.

Exposure Assessment of Essential and Potentially Toxic Metals in Wheat-Based Sweets for Human Consumption: Multivariate Analysis and Risk Evaluation Studies.

In recent years, there has been a growing concern about the negative impact of unforeseen contaminants such as metals in commonly consumed food items, which pose a threat to human well-being. Therefore, it is of utmost importance to evaluate the levels of these contaminants to guarantee the safe consumption of these food items. The goal of the current research is to determine the levels of essential (EMs: Mg, Ca, Mn, Fe, Co, Cu, and Zn) and potentially toxic metals (PTMs: Al, Cr, Ni, As, Cd, and Pb) in various brands of wheat-based sweets. One hundred samples were collected and analysed via flame atomic absorption spectrometry (FAAS) and inductively coupled plasma-optical emission spectrometry (ICP-OES). Also, the current study was to investigate the distribution, correlation, and multivariate analysis of 13 metals (Mg, Ca, Mn, Fe, Co, Cu, Zn, Al, Cr, Ni, As, Cd, and Pb). Hierarchical cluster analysis (HCA) and principal component analysis (PCA) were used to interpret the metals' association. The concentration (mg/kg) ranges of EMs were, in order, Mg (12.70-65.67), Ca (24.02-209.12), Mn (1.32-9.61), Fe (4.55-111.23), Co (0.32-8.94), Cu (2.12-8.61), and Zn (2.60-19.36), while the concentration (mg/kg) ranges of PTMs were, in order, Al (0.32-0.87), Cr (0.17-5.74), Ni (0.36-1.54), Cd (0.16-0.56), and Pb (0.14-0.92), and As was not detected in any sample under investigation. The HCA data revealed that Co, Al, and Ni form clusters with other metals. Sweets are prepared at high temperatures, and the elevated temperatures can increase the likelihood of Ni and Al leaching from stainless steel. Tolerable dietary intake (TDI) values for Ni were higher than the values established by the European Food Safety Authority (EFSA). The CR value found for the Ni and Cr was at the threshold level of cancer risk, if an amount of 25 g were to be used over a lifetime. In a nutshell, this study highlights the monitoring of EM and PTM levels in wheat-based sweets, and from a food safety perspective, the study is important for consumers of wheat-based sweets.

Influences of peanut hull-derived biochar, Trichoderma harzianum and supplemental phosphorus on hairy vetch growth in Pb- and Zn-contaminated soil.

In the present study, in order to improve the growth performance of hairy vetch (Vicia villosa Roth., Local landrace from Ardabil, Iran) seedlings grown in the soil contaminated with heavy metals Pb and Zn, our attention was directed toward the application of biochar, inoculation with conidial suspension of Trichoderma harzianum Rifai-T22 and management of phosphorus (P) nutrition. Heavy metal toxicity reduced leaf greenness, membrane stability index, maximum quantum yield of PSΙΙ (Fv/Fm), P concentration and uptake in plant tissues and root and shoot biomass, but increased Pb and Zn concentration and uptake in root and leaf, H2O2 and malondialdehyde content and CAT and POX activity in the leaves. The application of biochar, inoculation with Trichoderma fungus and P supplementation increased the shoot P content, which might contribute to the alleviation of P insufficiency and a subsequent elevation in P transfer to aboveground biomass, and eliminated the toxicity of heavy metal on hairy vetch plants, which was revealed in reducing oxidative stress and enhancing plant growth performance. The biochar considerably increased Zn immobilization, while being able to slightly stabilize Pb. Co-application of Trichoderma and 22 mg P/kg soil (22P) increased the concentration and uptake of Zn in the roots and decreased the translocation of this element to the shoots, especially when biochar was not amended. Although the biochar and P inputs could compensate the negative Trichoderma effects, the results suggested that biochar application in combination with fungal inoculation and 22-P supplementation could not only increase hairy vetch growth performance but also decline heavy metal uptake to ensure the production of a forage crop in soils polluted with heavy metals based on the nutritional standards of livestock.

Integral Study of Paramecium caudatum Acute and Chronic Toxicity, Sites of Entry and Distribution, Bioconcentration and Body Burdens of Five Metals.

An integral analysis of the acute and chronic toxicity, bioaccumulation, sites of entry, and distribution of four trace metals: copper, iron, lead, and nickel, and the non-trace metal mercury were performed in the ciliate Paramecium caudatum. Mercury was the fastest metal accumulated, and the most toxic. The sensitivity of Paramecium caudatum to the five metals tested (Cu, Fe, Hg, Ni, and Zn) falls in the range of other ciliate species. We observed similarities between the toxicity of the five metals to the ciliate P. caudatum with the rotifer Euchlanis dilatata: (a) Mercury was the most toxic metal in terms of acute and body burdens. (b) Acute values were very similar in both species, Hg as the most toxic and Fe as the less toxic, (c) the vacuole/ingestion chronic tests were more sensitive than growth inhibition chronic tests. These analyses would ideally help generate safer guidelines for protecting aquatic biota.

Pathological analysis of lesions in the exocrine pancreas of rats induced by Zinc Maltol.

The pancreas plays an important role in the homeostasis of zinc (Zn), a nutritionally essential metal. In several previous studies, Zn ions induced inflammatory changes in the exocrine pancreas; however, little is known about Zn complexes. In this study, we microscopically, immunohistochemically, and ultrastructurally examined pancreatic lesions in Sprague-Dawley (SD) rats induced by a 4-week repeated oral dose toxicity study of Zinc Maltol (ZM), a zinc (II) complex. ZM induces acinar atrophy and increases the number of duct-like structures. Immunohistochemistry revealed a decrease in the number of trypsin-positive cells, and an increase in the number of SOX9-positive cells. Interstitial fibrosis and macrophage infiltration also correlated with the degree of acinar atrophy. Electron microscopic evaluation revealed that the acinar cells that lost granules were surrounded by fibroblasts and collagen fibers. In conclusion, we provided a detailed description of ZM-induced pancreatic lesions in SD rats.

An updated overview on metal nanoparticles toxicity.

Although thousands of different nanoparticles (NPs) have been identified and synthesized to date, well-defined, consistent guidelines to control their exposure and evaluate their potential toxicity have yet to be fully established. As potential applications of nanotechnology in numerous fields multiply, there is an increased awareness of the issue of nanomaterials' toxicity among scientists and producers managing them. An updated inventory of customer products containing NPs estimates that they currently number over 5.000; ten years ago, they were one fifth of this. More often than not, products bear no information regarding the presence of NPs in the indicated list of ingredients or components. Consumers are therefore largely unaware of the extent to which nanomaterials have entered our lives, let alone their potential risks. Moreover, the lack of certainties with regard to the safe use of NPs is curbing their applications in the biomedical field, especially in the diagnosis and treatment of cancer, where they are performing outstandingly but are not yet being exploited as much as they could. The production of radical oxygen species is a predominant mechanism leading to metal NPs-driven carcinogenesis. The release of particularly reactive metal ions capable of crossing cell membranes has also been implicated in NPs toxicity. In this review we discuss the origin, behavior and biological toxicity of different metal NPs with the aim of rationalizing related health hazards and calling attention to toxicological concerns involved in their increasingly widespread use.

Phytochelatin-mediated metal detoxification pathway is crucial for an organomercurial phenylmercury tolerance in Arabidopsis.

KEY MESSAGE: An organomercurial phenylmercury activates AtPCS1, an enzyme known for detoxification of inorganic metal(loid) ions in Arabidopsis and the induced metal-chelating peptides phytochelatins are essential for detoxification of phenylmercury. Small thiol-rich peptides phytochelatins (PCs) and their synthases (PCSs) are crucial for plants to mitigate the stress derived from various metal(loid) ions in their inorganic form including inorganic mercury [Hg(II)]. However, the possible roles of the PC/PCS system in organic mercury detoxification in plants remain elusive. We found that an organomercury phenylmercury (PheHg) induced PC synthesis in Arabidopsis thaliana plants as Hg(II), whereas methylmercury did not. The analyses of AtPCS1 mutant plants and in vitro assays using the AtPCS1-recombinant protein demonstrated that AtPCS1, the major PCS in A. thaliana, was responsible for the PheHg-responsive PC synthesis. AtPCS1 mutants cad1-3 and cad1-6, and the double mutant of PC-metal(loid) complex transporters AtABCC1 and AtABCC2 showed enhanced sensitivity to PheHg as well as to Hg(II). The hypersensitivity of cad1-3 to PheHg stress was complemented by the own-promoter-driven expression of AtPCS1-GFP. The confocal microscopy of the complementation lines showed that the AtPCS1-GFP was preferentially expressed in epidermal cells of the mature and elongation zones, and the outer-most layer of the lateral root cap cells in the meristematic zone. Moreover, in vitro PC-metal binding assay demonstrated that binding affinity between PC and PheHg was comparable to Hg(II). However, plant ionomic profiles, as well as root morphology under PheHg and Hg(II) stress, were divergent. These results suggest that PheHg phytotoxicity is different from Hg(II), but AtPCS1-mediated PC synthesis, complex formation, and vacuolar sequestration by AtABCC1 and AtABCC2 are similarly functional for both PheHg and Hg(II) detoxification in root surficial cell types.

Construction and analysis of a Noccaea caerulescens TILLING population.

BACKGROUND: Metals such as Zn or Cd are toxic to plant and humans when they are exposed in high quantities through contaminated soil or food. Noccaea caerulescens, an extraordinary Zn/Cd/Ni hyperaccumulating species, is used as a model plant for metal hyperaccumulation and phytoremediation studies. Current reverse genetic techniques to generate mutants based on transgenesis is cumbersome due to the low transformation efficiency of this species. We aimed to establish a mutant library for functional genomics by a non-transgenic approach, to identify mutants with an altered mineral profiling, and to screen for mutations in bZIP19, a regulator of Zn homeostasis in N. caerulescens. RESULTS: To generate the N. caerulescens mutant library, 3000 and 5000 seeds from two sister plants of a single-seed recurrent inbred descendant of the southern French accession Saint-Félix-de-Pallières (SF) were mutagenized respectively by 0.3 or 0.4% ethyl methane sulfonate (EMS). Two subpopulations of 5000 and 7000 M2 plants were obtained after 0.3 or 0.4% EMS treatment. The 0.4% EMS treatment population had a higher mutant frequency and was used for TILLING. A High Resolution Melting curve analysis (HRM) mutation screening platform was optimized and successfully applied to detect mutations for NcbZIP19, encoding a transcription factor controlling Zn homeostasis. Of four identified point mutations in NcbZIP19, two caused non-synonymous substitutions, however, these two mutations did not alter the ionome profile compared to the wild type. Forward screening of the 0.4% EMS treatment population by mineral concentration analysis (ionomics) in leaf material of each M2 plant revealed putative mutants affected in the concentration of one or more of the 20 trace elements tested. Several of the low-Zn mutants identified in the ionomic screen did not give progeny, illustrating the importance of Zn for the species. The mutant frequency of the population was evaluated based on an average of 2.3 knockout mutants per tested monogenic locus. CONCLUSIONS: The 0.4% EMS treatment population is effectively mutagenized suitable for forward mutant screens and TILLING. Difficulties in seed production in low Zn mutants, obtained by both forward and reverse genetic approach, hampered further analysis of the nature of the low Zn phenotypes.

Melatonin as a regulator of plant ionic homeostasis: implications for abiotic stress tolerance.

Melatonin is a highly conserved and ubiquitous molecule that operates upstream of a broad array of receptors in animal systems. Since melatonin was discovered in plants in 1995, hundreds of papers have been published revealing its role in plant growth, development, and adaptive responses to the environment. This paper summarizes the current state of knowledge of melatonin's involvement in regulating plant ion homeostasis and abiotic stress tolerance. The major topics covered here are: (i) melatonin's control of H+-ATPase activity and its implication for plant adaptive responses to various abiotic stresses; (ii) regulation of the reactive oxygen species (ROS)-Ca2+ hub by melatonin and its role in stress signaling; and (iii) melatonin's regulation of ionic homeostasis via hormonal cross-talk. We also show that the properties of the melatonin molecule allow its direct scavenging of ROS, thus preventing negative effects of ROS-induced activation of ion channels. The above 'desensitization' may play a critical role in preventing stress-induced K+ loss from the cytosol as well as maintaining basic levels of cytosolic Ca2+ required for optimal cell operation. Future studies should focus on revealing the molecular identity of transporters that could be directly regulated by melatonin and providing a bioinformatic analysis of evolutionary aspects of melatonin sensing and signaling.

Metal crossroads in plants: modulation of nutrient acquisition and root development by essential trace metals.

The metals iron, zinc, manganese, copper, molybdenum, and nickel are essential for the growth and development of virtually all plant species. Although these elements are required at relatively low amounts, natural factors and anthropogenic activities can significantly affect their availability in soils, inducing deficiencies or toxicities in plants. Because essential trace metals can shape root systems and interfere with the uptake and signaling mechanisms of other nutrients, the non-optimal availability of any of them can induce multi-element changes in plants. Interference by one essential trace metal with the acquisition of another metal or a non-metal nutrient can occur prior to or during root uptake. Essential trace metals can also indirectly impact the plant's ability to capture soil nutrients by targeting distinct root developmental programs and hormone-related processes, consequently inducing largely metal-specific changes in root systems. The presence of metal binding domains in many regulatory proteins also enables essential trace metals to coordinate nutrient uptake by acting at high levels in hierarchical signaling cascades. Here, we summarize the known molecular and cellular mechanisms underlying trace metal-dependent modulation of nutrient acquisition and root development, and highlight the importance of considering multi-element interactions to breed crops better adapted to non-optimal trace metal availabilities.