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Foliar fertilization delivers essential nutrients directly to plant tissues, reducing excessive soil fertilizer applications that can lead to eutrophication following nutrient leaching. Foliar nutrient absorption is a dynamic process affected by leaf surface structure and composition, plant nutrient status, and ion physicochemical properties. We applied multiple methods to study the foliar absorption behaviors of manganese (Mn) and phosphorus (P) in nutrient-deficient spring barley (Hordeum vulgare) at two growth stages. Nutrient-specific chlorophyll a fluorescence assays were used to visualize leaf nutrient status, while laser ablation-inductively coupled plasma-mass spectrometry (LA-ICP-MS) was used to visualize foliar absorption pathways for P and Mn ions. Rapid Mn absorption was facilitated by a relatively thin cuticle with a low abundance of waxes and a higher stomatal density in Mn-deficient plants. Following absorption, Mn accumulated in epidermal cells and in the photosynthetically active mesophyll, enabling a fast (6 h) restoration of Mn-dependent photosynthetic processes. Conversely, P-deficient plants developed thicker cuticles and epidermal cell walls, which reduced the penetration of P across the leaf surface. Foliar-applied P accumulated in trichomes and fiber cells above leaf veins without reaching the mesophyll and, as a consequence, no restoration of P-dependent photosynthetic processes was observed. This study reveals new links between leaf surface morphology, foliar-applied ion absorption pathways, and the restoration of affected physiological processes in nutrient-deficient leaves. Understanding that ions may have different absorption pathways across the leaf surface is critical for the future development of efficient fertilization strategies for crops in nutrient-limited soils.
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Hordeum , Manganês , Fósforo , Folhas de Planta , Clorofila A/análise , Hordeum/metabolismo , Íons/metabolismo , Manganês/metabolismo , Nutrientes/análise , Fósforo/metabolismo , Folhas de Planta/metabolismo , SoloRESUMO
Increasing atmospheric CO2 concentration is expected to enhance the grain yield of C3 cereal plants, while at the same time reducing the concentrations of minerals and proteins. This will lead to a lower nutritional quality and increase global problems associated with micronutrient malnutrition. Among the barley grain storage proteins, the C-hordein fraction has the lowest abundance of sulfur (S) containing amino acids and is poorest in binding of zinc (Zn). In the present study, C-hordein-suppressed barley lines with reduced C-hordein content, obtained by use of antisense or RNAi technology, were investigated under ambient and elevated atmospheric CO2 concentration. Grains of the C-hordein-suppressed lines showed 50% increase in the concentrations of Zn and iron (Fe) in the core endosperm relative to the wild-type under both ambient and elevated atmospheric CO2 . Element distribution images obtained using laser ablation-inductively coupled plasma-mass spectrometry confirmed the enrichment of Fe and Zn in the core endosperm of the lines with modified storage protein composition. We conclude that modification of grain storage proteins may improve the nutritional value of cereal grain with respect to Zn and Fe under both normal and future conditions of elevated atmospheric CO2 .
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Endosperma , Hordeum , Dióxido de Carbono/metabolismo , Grão Comestível/metabolismo , Hordeum/metabolismo , Ferro/metabolismo , Zinco/metabolismoRESUMO
Manganese (Mn) plays an important role in the oxygen-evolving complex, where energy from light absorption is used for water splitting. Although changes in light intensity and Mn status can interfere with the functionality of the photosynthetic apparatus, the interaction between these two factors and the underlying mechanisms remain largely unknown. Here, maize seedlings were grown hydroponically and exposed to two different light intensities under Mn-sufficient or -deficient conditions. No visual Mn deficiency symptoms appeared even though the foliar Mn concentration in the Mn-deficient treatments was reduced to 2 µg g-1. However, the maximum quantum yield efficiency of PSII and the net photosynthetic rate declined significantly, indicating latent Mn deficiency. The reduction in photosynthetic performance by Mn depletion was further aggravated when plants were exposed to high light intensity. Integrated transcriptomic and proteomic analyses showed that a considerable number of genes encoding proteins in the photosynthetic apparatus were only suppressed by a combination of Mn deficiency and high light, thus indicating interactions between changes in Mn nutritional status and light intensity. We conclude that high light intensity aggravates latent Mn deficiency in maize by interfering with the abundance of PSII proteins.
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Manganês , Zea mays , Luz , Fotossíntese , Complexo de Proteína do Fotossistema II/metabolismo , Proteômica , Zea mays/genética , Zea mays/metabolismoRESUMO
The broad variability of Cucumis melo (melon, Cucurbitaceae) presents a challenge to conventional classification and organization within the species. To shed further light on the infraspecific relationships within C. melo, we compared genotypic and metabolomic similarities among 44 accessions representative of most of the cultivar-groups. Genotyping-by-sequencing (GBS) provided over 20,000 single-nucleotide polymorphisms (SNPs). Metabolomics data of the mature fruit flesh and rind provided over 80,000 metabolomic and elemental features via an orchestra of six complementary metabolomic platforms. These technologies probed polar, semi-polar, and non-polar metabolite fractions as well as a set of mineral elements and included both flavor- and taste-relevant volatile and non-volatile metabolites. Together these results enabled an estimate of "metabolomic/elemental distance" and its correlation with the genetic GBS distance of melon accessions. This study indicates that extensive and non-targeted metabolomics/elemental characterization produced classifications that strongly, but not completely, reflect the current and extensive genetic classification. Certain melon Groups, such as Inodorous, clustered in parallel with the genetic classifications while other genome to metabolome/element associations proved less clear. We suggest that the combined genomic, metabolic, and element data reflect the extensive sexual compatibility among melon accessions and the breeding history that has, for example, targeted metabolic quality traits, such as taste and flavor.
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BACKGROUND: To understand processes regulating nutrient homeostasis at the single-cell level there is a need for new methods that allow multi-element profiling of biological samples ultimately only available as isolated tissues or cells, typically in nanogram-sized samples. Apart from tissue isolation, the main challenges for such analyses are to obtain a complete and homogeneous digestion of each sample, to keep sample dilution at a minimum and to produce accurate and reproducible results. In particular, determining the weight of small samples becomes increasingly challenging when the sample amount decreases. RESULTS: We developed a novel method for sampling, digestion and multi-element analysis of nanogram-sized plant tissue, along with strategies to quantify element concentrations in samples too small to be weighed. The method is based on tissue isolation by laser capture microdissection (LCM), followed by pressurized micro-digestion and ICP-MS analysis, the latter utilizing a stable µL min-1 sample aspiration system. The method allowed for isolation, digestion and analysis of micro-dissected tissues from barley roots with an estimated sample weight of only ~ 400 ng. In the collection and analysis steps, a number of contamination sources were identified. Following elimination of these sources, several elements, including magnesium (Mg), phosphorus (P), potassium (K) and manganese (Mn), could be quantified. By measuring the exact area and thickness of each of the micro-dissected tissues, their volume was calculated. Combined with an estimated sample density, the sample weights could subsequently be calculated and the fact that these samples were too small to be weighed could thereby be circumvented. The method was further documented by analysis of Arabidopsis seeds (~ 20 µg) as well as tissue fractions of such seeds (~ 10 µg). CONCLUSIONS: The presented method enables collection and multi-element analysis of small-sized biological samples, ranging down to the nanogram level. As such, the method paves the road for single cell and tissue-specific quantitative ionomics, which allow for future transcriptional, proteomic and metabolomic data to be correlated with ionomic profiles. Such analyses will deepen our understanding of how the elemental composition of plants is regulated, e.g. by transporter proteins and physical barriers (i.e. the Casparian strip and suberin lamellae in the root endodermis).
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Tryptophan is a key component in many biological processes and an essential amino acid in food and feed materials. Analysis of the tryptophan content in proteins or protein-containing matrices has always been a challenge. We show here that the preparation of samples prior to tryptophan analysis can be significantly simplified, and the time consumption reduced, by using ascorbic acid as antioxidant to eliminate the problem of tryptophan degradation during alkaline hydrolysis. Combined with separation by HPLC and detection by Single Quadrupole Mass Spectrometry, this allows the analytical run time to be reduced to 10 min. The alkaline hydrolysate obtained in the method presented here may be combined with the oxidized hydrolysate obtained when sulfur-containing amino acids are to be measured, thus essentially providing two analyses for the time of one.
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Plants have evolved different strategies to utilize various forms of nitrogen (N) from the environment. While regulation of plant growth and development in response to application of inorganic N forms has been characterized, our knowledge about the effect on cell wall structure and composition is quite limited. In this study, we analysed cell walls of Brachypodium distachyon supplied with three types of inorganic N (NH4NO3, NO3-, or NH4+). Cell wall profiles showed distinct alterations in both the quantity and structures of individual polymers. Nitrate stimulated cellulose, but inhibited lignin deposition at the heading growth stage. On the other hand, ammonium supply resulted in higher concentration of mixed linkage glucans. In addition, the chemical structure of pectins and hemicelluloses was strongly influenced by the form of N. Supply of only NO3- led to alteration in xylan substitution and to lower esterification of homogalacturonan. We conclude that the physiological response to absorption of different inorganic N forms includes pleotropic remodelling of type II cell walls.
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Brachypodium/metabolismo , Parede Celular/metabolismo , Nitrogênio/farmacologia , Compostos de Amônio/metabolismo , Biomassa , Brachypodium/efeitos dos fármacos , Brachypodium/crescimento & desenvolvimento , Parede Celular/efeitos dos fármacos , Celulose/metabolismo , Epitopos/metabolismo , Esterificação , Glucanos/metabolismo , Lignina/metabolismo , Nitratos/farmacologia , Raízes de Plantas/efeitos dos fármacos , Raízes de Plantas/metabolismo , Brotos de Planta/efeitos dos fármacos , Brotos de Planta/metabolismoRESUMO
KEY MESSAGE: This study demonstrates that an active breeding nursery with rotation can be used to identify marker-trait associations for biomass yield and quality parameters that are important for biorefinery purposes. Wheat straw is a valuable feedstock for bioethanol production, but due to the recalcitrant nature of lignocellulose, its efficient use in biorefineries is limited by its low digestibility and difficult conversion of structural carbohydrates into free sugars. A genome-wide association study (GWAS) was conducted to search for significant SNP markers that could be used in a breeding programme to improve the value of wheat straw in a biorefinery setting. As part of a 3-year breeding programme (2013-2016), 190 winter wheat lines were phenotyped for traits that affect the yield and quality of the harvested biomass. These traits included straw yield, plant height, lodging at three growth stages and Septoria tritici blotch (STB) susceptibility. Release of glucose, xylose and arabinose was determined after hydrothermal pretreatment and enzymatic hydrolysis of the straw. The lines were genotyped using 15 K SNP markers and 5552 SNP markers could be used after filtering. Heritability for all traits ranged from 0.02 to 0.74. GWASs were conducted using CMLM, SUPER and FarmCPU algorithms, to analyse which algorithm could detect the highest number of marker-trait associations (MTAs). Comparable tendencies were obtained from CMLM and FarmCPU, but FarmCPU produced the most significant results. MTAs were obtained for lodging, harvest index, plant height, STB, glucose, xylose and arabinose at a significance level of p < 9.01 × 10-6. MTAs in chromosome 6A were observed for glucose, xylose and arabinose, and could be of importance for increasing sugar release for bioethanol production.
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Melhoramento Vegetal , Característica Quantitativa Herdável , Triticum/crescimento & desenvolvimento , Triticum/genética , Biomassa , Estudos de Associação Genética , Marcadores Genéticos , Genótipo , Fenótipo , Polimorfismo de Nucleotídeo ÚnicoRESUMO
Manganese (Mn) deficiency affects various processes in plant shoots. However, the functions of Mn in roots and the processes involved in root adaptation to Mn deficiency are largely unresolved. Here, we show that the suberization of endodermal cells in barley (Hordeum vulgare) roots is altered in response to Mn deficiency, and that the intensity of Mn deficiency ultimately determines whether suberization increases or decreases. Mild Mn deficiency increased the length of the unsuberized zone close to the root tip, and increased the distance from the root tip at which the fully suberized zone developed. By contrast, strong Mn deficiency increased suberization closer to the root tip. Upon Mn resupply, suberization was identical to that seen on Mn-replete plants. Bioimaging and xylem sap analyses suggest that the reduced suberization in mildly Mn-deficient plants promotes radial Mn transport across the endodermis at a greater distance from the root tip. Less suberin also favors the inwards radial transport of calcium and sodium, but negatively affects the potassium concentration in the stele. During strong Mn deficiency, Mn uptake was directed toward the root tip. Enhanced suberization provides a mechanism to prevent absorbed Mn from leaking out of the stele. With more suberin, the inward radial transport of calcium and sodium decreases, whereas that of potassium increases. We conclude that changes in suberization in response to the intensity of Mn deficiency have a strong effect on root ion homeostasis and ion translocation.
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Hordeum/metabolismo , Lipídeos , Manganês/metabolismo , Raízes de Plantas/metabolismo , Homeostase , Hordeum/crescimento & desenvolvimento , Íons/metabolismo , Espectrometria de Massas/métodosRESUMO
Cytosolic glutamine synthetase (GS1) plays a central role in nitrogen (N) metabolism. The importance of GS1 in N remobilization during reproductive growth has been reported in cereal species but attempts to improve N utilization efficiency (NUE) by overexpressing GS1 have yielded inconsistent results. Here, we demonstrate that transformation of barley (Hordeum vulgare L.) plants using a cisgenic strategy to express an extra copy of native HvGS1-1 lead to increased HvGS1.1 expression and GS1 enzyme activity. GS1 overexpressing lines exhibited higher grain yields and NUE than wild-type plants when grown under three different N supplies and two levels of atmospheric CO2 . In contrast with the wild-type, the grain protein concentration in the GS1 overexpressing lines did not decline when plants were exposed to elevated (800-900 µL/L) atmospheric CO2 . We conclude that an increase in GS1 activity obtained through cisgenic overexpression of HvGS1-1 can improve grain yield and NUE in barley. The extra capacity for N assimilation obtained by GS1 overexpression may also provide a means to prevent declining grain protein levels under elevated atmospheric CO2 .
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Dióxido de Carbono/química , Glutamato-Amônia Ligase/metabolismo , Proteínas de Grãos/metabolismo , Hordeum/metabolismo , Nitrogênio/metabolismo , Regulação da Expressão Gênica de Plantas , Glutamato-Amônia Ligase/genética , Hordeum/genética , Plantas Geneticamente Modificadas/metabolismoRESUMO
Foliar application of zinc (Zn) to crops is an effective way to increase the grain concentration of Zn. However, the development of more efficient foliar Zn fertilizers is limited by a lack of knowledge regarding the distribution, mobility, and speciation of Zn in leaves once it is taken up by the plant. We performed an experiment using radiolabelled Zn (65Zn), and in situ time-resolved elemental imaging using synchrotron X-ray fluorescence microscopy (XFM), to investigate the behaviour of two commonly used Zn foliar fertilizers (Zn sulphate and ZnEDTA) in wheat (Triticum aestivum) leaves. Both experiments showed that Zn had limited mobility in leaves, moving <25 mm from the application point after 24 h. Although limited, the translocation of Zn occurred quickly for both treatments; moving more between 3 h and 12 h after application than between 12 h and 24 h. Speciation analysis using synchrotron-based X-ray absorption near-edge structure (XANES) showed that ZnEDTA was in fact taken up in chelated form and not as ionic Zn (Zn2+). The XANES data also showed that Zn, from both treatments, was then complexed by ligands in the leaf (e.g. phytate and citrate), potentially in response to localized Zn toxicity. The results of the present study provide important insights into the behaviour of commonly used foliar-applied Zn fertilizers, and can be used to optimize current fertilization strategies and contribute to the development of more efficient foliar Zn fertilizers.
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Ácido Edético/farmacocinética , Fertilizantes/análise , Folhas de Planta/metabolismo , Triticum/efeitos dos fármacos , Sulfato de Zinco/farmacocinética , Zinco/farmacocinética , Transporte Biológico , Grão Comestível/química , Grão Comestível/efeitos dos fármacos , Folhas de Planta/efeitos dos fármacos , Triticum/metabolismo , Espectroscopia por Absorção de Raios XRESUMO
This special issue of Physiologia Plantarum has been prepared to publish some of the highlights presented at the XVIII International Plant Nutrition Colloquium (IPNC), which took place during 19-24 August, 2017 in Copenhagen, Denmark. The meeting was organized by University of Copenhagen. Close to 600 participants from more than 50 different countries participated. This article is protected by copyright. All rights reserved.
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BACKGROUND: The amino acid profile of plants is an important parameter in assessments of their growth potential, resource-use efficiency and/or quality as food and feed. Screening studies may involve large number of samples but the classical amino acid analysis is limited by the fact that it is very time consuming with typical chromatographic run times of 70 min or more. RESULTS: We have here developed a high-throughput method for analysis of amino acid profiles in plant materials. The method combines classical protein hydrolysis and derivatization with fast separation by UHPLC and detection by a single quadrupole (QDa) mass spectrometer. The chromatographic run time is reduced to 10 min and the precision, accuracy and sensitivity of the method are in line with other recent methods utilizing advanced and more expensive mass spectrometers. The sensitivity of the method is at least a factor 10 better than that of methods relying on detection by fluorescence or UV. It is possible to downscale sample size to 20 mg without compromising reproducibility, which makes the method ideal for analysis of very small sample amounts. CONCLUSION: The developed method allows high-throughput analysis of amino acid profiles in plant materials. The analysis is robust and accurate as well as compatible with both free amino acids and protein hydrolysates. The QDa detector offers high sensitivity and accuracy, while at the same time being relatively simple to operate and cheap to purchase, thus significantly reducing the overall analytical costs compared to methods based on more advanced mass spectrometers.
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Silicon (Si) has many beneficial effects in plants, especially for the survival from biotic and abiotic stresses. However, Si may negatively affect the quality of lignocellulosic biomass for bioenergy purposes. Despite many studies, the regulation of Si distribution and deposition in plants remains to be fully understood. Here, we have identified the Brachypodium distachyon mutant low-silicon 1 (Bdlsi1-1), with impaired channeling function of the Si influx transporter BdLSI1, resulting in a substantial reduction of Si in shoots. Bioimaging by laser ablation-inductively coupled plasma-mass spectrometry showed that the wild-type plants deposited Si mainly in the bracts, awns and leaf macrohairs. The Bdlsi1-1 mutants showed substantial (>90%) reduction of Si in the mature shoots. The Bdlsi1-1 leaves had fewer, shorter macrohairs, but the overall pattern of Si distribution in bracts and leaf tissues was similar to that in the wild-type. The Bdlsi1-1 plants supplied with Si had significantly lower seed weights, compared to the wild-type. In low-Si media, the seed weight of wild-type plants was similar to that of Bdlsi1-1 mutants supplied with Si, while the Bdlsi1-1 seed weight decreased further. We conclude that Si deficiency results in widespread alterations in leaf surface morphology and seed formation in Brachypodium, showing the importance of Si for successful development in grasses.
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Brachypodium/efeitos dos fármacos , Proteínas de Membrana Transportadoras/metabolismo , Silício/farmacologia , Brachypodium/crescimento & desenvolvimento , Proteínas de Membrana Transportadoras/genética , Mutação , Folhas de Planta/efeitos dos fármacos , Folhas de Planta/crescimento & desenvolvimento , Sementes/efeitos dos fármacos , Sementes/crescimento & desenvolvimentoRESUMO
Transporters involved in manganese (Mn) uptake and intracellular Mn homeostasis in Arabidopsis and rice are well characterized, while much less is known for barley, which is particularly prone to Mn deficiency. In this study we have investigated the role of the iron-regulated transporter 1 (IRT1) for Mn uptake and translocation in barley plants. We employed an RNAi approach to reduce HvIRT1 expression to 5% of the wild-type level. This enabled characterization of the functional role of HvIRT1 by use of advanced imaging and phenotyping techniques applied to plants growing in hydroponics or soils with different Mn availability. Our results highlight the importance of HvIRT1 for the transport of Mn across the root endodermis into the stele. In the hvirt1-RNAi lines, a chlorotic phenotype with reduced shoot Mn concentration and impaired photosynthetic functionality was observed, especially under conditions with low Mn availability. We also document that HvIRT1 controlled the Mn distribution within the barley grain. Surprisingly, unlike other IRT1 orthologues, HvIRT1 played no significant role in iron uptake. We conclude that the barley IRT1 orthologue has a novel function with respect to ensuring sufficient shoot Mn concentrations. The preference of IRT1 for Mn instead of Fe is discussed in an evolutionary context.
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Hordeum/metabolismo , Ferro/metabolismo , Manganês/metabolismo , Proteínas de Plantas/metabolismo , Transporte Biológico , Regulação da Expressão Gênica de Plantas , Hordeum/genética , Modelos Biológicos , Fenótipo , Proteínas de Plantas/genética , Raízes de Plantas/metabolismo , Brotos de Planta/metabolismo , Interferência de RNA , Sementes/metabolismo , Xilema/metabolismoRESUMO
A novel zinc (Zn) fertilizer concept based on Zn-doped layered double hydroxides (Zn-doped Mg-Fe-LDHs) has been investigated. Zn-doped Mg-Fe-LDHs were synthesized, their chemical composition was analyzed, and their nutrient release was studied in buffered solutions with different pH values. Uptake of Zn by barley (Hordeum vulgare cv. Antonia) was evaluated in short- (8 weeks), medium- (11 weeks), and long-term (28 weeks) experiments in quartz sand and in a calcareous soil enriched with Zn-doped Mg-Fe-LDHs. The Zn release rate of the Zn-doped Mg-Fe-LDHs was described by a first-order kinetics equation showing maximum release at pH 5.2, reaching approximately 45% of the total Zn content. The Zn concentrations in the plants receiving the LDHs were between 2- and 9.5-fold higher than those in plants without Zn addition. A positive effect of the LDHs was also found in soil. This work documents the long-term Zn release capacity of LDHs complying with a release-on-demand behavior and serves as proof-of-concept that Zn-doped Mg-Fe-LDHs can be used as Zn fertilizers.
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Fertilizantes/análise , Hordeum/metabolismo , Hidróxidos/química , Zinco/metabolismo , Hordeum/crescimento & desenvolvimento , Hidróxidos/metabolismo , Cinética , Zinco/químicaRESUMO
Basic leucine zipper (bZIP) transcription factors control important developmental and physiological processes in plants. In Arabidopsis thaliana, the three gene F-bZIP subfamily has been associated with zinc deficiency and salt stress response. Benefiting from the present abundance of plant genomic data, we performed an evolutionary and structural characterization of plant F-bZIPs. We observed divergence during seed plant evolution, into two groups and inferred different selective pressures for each. Group 1 contains AtbZIP19 and AtbZIP23 and appears more conserved, whereas Group 2, containing AtbZIP24, is more prone to gene loss and expansion events. Transcriptomic and experimental data reinforced AtbZIP19/23 as pivotal regulators of the zinc deficiency response, mostly via the activation of genes from the ZIP metal transporter family, and revealed that they are the main regulatory switch of AtZIP4. A survey of AtZIP4 orthologs promoters across different plant taxa revealed an enrichment of the Zinc Deficiency Response Element (ZDRE) to which both AtbZIP19/23 bind. Overall, our results indicate that while the AtbZIP24 function in the regulation of the salt stress response may be the result of neo-functionalization, the AtbZIP19/23 function in the regulation of the zinc deficiency response may be conserved in land plants (Embryophytes).
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Proteínas de Arabidopsis/genética , Arabidopsis/genética , Zíper de Leucina , Filogenia , Fatores de Transcrição/genética , Zinco/deficiência , Arabidopsis/metabolismo , Proteínas de Arabidopsis/metabolismo , Fatores de Transcrição/metabolismo , Zinco/metabolismoRESUMO
A catalytic manganese (Mn) cluster is required for the oxidation of water in the oxygen-evolving complex (OEC) of photosystem II (PSII) in plants. Despite this essential role of Mn in generating the electrons driving photosynthesis, limited information is available on how Mn deficiency affects PSII functionality. We have here used parameters derived from measurements of fluorescence induction kinetics (OJIP transients), non-photochemical quenching (NPQ) and PSII subunit composition to investigate how latent Mn deficiency changes the photochemistry in two barley genotypes differing in Mn efficiency. Mn deficiency caused dramatic reductions in the quantum yield of PSII and led to the appearance of two new inflection points, the K step and the D dip, in the OJIP fluorescence transients, indicating severe damage to the OEC. In addition, Mn deficiency decreased the ability to induce NPQ in the light, rendering the plants incapable of dissipating excess energy in a controlled way. Thus, the Mn deficient plants became severely affected in their ability to recover from high light-induced photoinhibition, especially under strong Mn deficiency. Interestingly, the Mn-efficient genotype was able to maintain a higher NPQ than the Mn-inefficient genotype when exposed to mild Mn deficiency. However, during severe Mn deficiency, there were no differences between the two genotypes, suggesting a general loss of the ability to disassemble and repair PSII. The pronounced defects of PSII activity were supported by a dramatic decrease in the abundance of the OEC protein subunits, PsbP and PsbQ in response to Mn deficiency for both genotypes. We conclude that regulation of photosynthetic performance by means of maintaining and inducing NPQ mechanisms contribute to genotypic differences in the Mn efficiency of barley genotypes growing under conditions with mild Mn deficiency.
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Cytosolic glutamine synthetase 1;2 plays an important role in the primary nitrogen assimilation in roots. Based on characterization of the knockout mutant gln1;2 we have recently demonstrated that Gln1;2 is also essential for ammonium handling in shoots. Here we built reciprocally grafted plants between wild type (Wt) and gln1;2 in order to separate the root and shoot roles of Gln1;2. Significant reduction in silique number and seed yield were observed in the grafted plants 1;2shoot/Wtroot relative to Wtshoot/1;2root and Wtshoot/Wtroot. Shoot Gln1;2 thus played a crucial role for seed production. Tracing experiments with 15N showed that the relative nitrogen remobilization from vegetative organs to seeds in gln1;2 was just as efficient as in the Wt plants. This was the case although the total quantity of nitrogen in gln1;2 was significantly lower compared to that in the Wt. We conclude that the functions of shoot Gln1;2 are primarily associated with internal N signaling for establishment of seed yield capacity rather than with nitrogen remobilization.
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Citosol/enzimologia , Glutamato-Amônia Ligase/metabolismo , Raízes de Plantas/enzimologia , Raízes de Plantas/metabolismo , Brotos de Planta/enzimologia , Brotos de Planta/metabolismo , Arabidopsis/enzimologia , Arabidopsis/metabolismo , Regulação da Expressão Gênica de Plantas , Nitrogênio/metabolismo , Sementes/enzimologia , Sementes/metabolismoRESUMO
Better understanding of root function is central for the development of plants with more efficient nutrient uptake and translocation. We here present a method for multielement bioimaging at the cellular level in roots of the genetic model system Arabidopsis (Arabidopsis thaliana). Using conventional protocols for microscopy, we observed that diffusible ions such as potassium and sodium were lost during sample dehydration. Thus, we developed a protocol that preserves ions in their native, cellular environment. Briefly, fresh roots are encapsulated in paraffin, cryo-sectioned, and freeze dried. Samples are finally analyzed by laser ablation-inductively coupled plasma-mass spectrometry, utilizing a specially designed internal standard procedure. The method can be further developed to maintain the native composition of proteins, enzymes, RNA, and DNA, making it attractive in combination with other omics techniques. To demonstrate the potential of the method, we analyzed a mutant of Arabidopsis unable to synthesize the metal chelator nicotianamine. The mutant accumulated substantially more zinc and manganese than the wild type in the tissues surrounding the vascular cylinder. For iron, the images looked completely different, with iron bound mainly in the epidermis of the wild-type plants but confined to the cortical cell walls of the mutant. The method offers the power of inductively coupled plasma-mass spectrometry to be fully employed, thereby providing a basis for detailed studies of ion transport in roots. Being applicable to Arabidopsis, the molecular and genetic approaches available in this system can now be fully exploited in order to gain a better mechanistic understanding of these processes.