Plant polygalacturonase structures specify enzyme dynamics and processivities to fine-tune cell wall pectins.

Polygalacturonases (PGs) fine-tune pectins to modulate cell wall chemistry and mechanics, impacting plant development. The large number of PGs encoded in plant genomes leads to questions on the diversity and specificity of distinct isozymes. Herein, we report the crystal structures of 2 Arabidopsis thaliana PGs, POLYGALACTURONASE LATERAL ROOT (PGLR), and ARABIDOPSIS DEHISCENCE ZONE POLYGALACTURONASE2 (ADPG2), which are coexpressed during root development. We first determined the amino acid variations and steric clashes that explain the absence of inhibition of the plant PGs by endogenous PG-inhibiting proteins (PGIPs). Although their beta helix folds are highly similar, PGLR and ADPG2 subsites in the substrate binding groove are occupied by divergent amino acids. By combining molecular dynamic simulations, analysis of enzyme kinetics, and hydrolysis products, we showed that these structural differences translated into distinct enzyme-substrate dynamics and enzyme processivities: ADPG2 showed greater substrate fluctuations with hydrolysis products, oligogalacturonides (OGs), with a degree of polymerization (DP) of ≤4, while the DP of OGs generated by PGLR was between 5 and 9. Using the Arabidopsis root as a developmental model, exogenous application of purified enzymes showed that the highly processive ADPG2 had major effects on both root cell elongation and cell adhesion. This work highlights the importance of PG processivity on pectin degradation regulating plant development.

Magnesium and calcium overaccumulate in the leaves of a schengen3 mutant of Brassica rapa.

Magnesium (Mg) and calcium (Ca) are essential mineral nutrients poorly supplied in many human food systems. In grazing livestock, Mg and Ca deficiencies are costly welfare issues. Here, we report a Brassica rapa loss-of-function schengen3 (sgn3) mutant, braA.sgn3.a-1, which accumulates twice as much Mg and a third more Ca in its leaves. We mapped braA.sgn3.a to a single recessive locus using a forward ionomic screen of chemically mutagenized lines with subsequent backcrossing and linked-read sequencing of second back-crossed, second filial generation (BC2F2) segregants. Confocal imaging revealed a disrupted root endodermal diffusion barrier, consistent with SGN3 encoding a receptor-like kinase required for normal formation of Casparian strips, as reported in thale cress (Arabidopsis thaliana). Analysis of the spatial distribution of elements showed elevated extracellular Mg concentrations in leaves of braA.sgn3.a-1, hypothesized to result from preferential export of excessive Mg from cells to ensure suitable cellular concentrations. This work confirms a conserved role of SGN3 in controlling nutrient homeostasis in B. rapa, and reveals mechanisms by which plants are able to deal with perturbed shoot element concentrations resulting from a "leaky" root endodermal barrier. Characterization of variation in leaf Mg and Ca accumulation across a mutagenized population of B. rapa shows promise for using such populations in breeding programs to increase edible concentrations of essential human and animal nutrients.

Effect of phosphorus supply on root traits of two Brassica oleracea L. genotypes.

BACKGROUND: Phosphorus (P) deficiency limits crop production worldwide. Crops differ in their ability to acquire and utilise the P available. The aim of this study was to determine root traits (root exudates, root system architecture (RSA), tissue-specific allocation of P, and gene expression in roots) that (a) play a role in P-use efficiency and (b) contribute to large shoot zinc (Zn) concentration in Brassica oleracea. RESULTS: Two B. oleracea accessions (var. sabellica C6, a kale, and var. italica F103, a broccoli) were grown in a hydroponic system or in a high-throughput-root phenotyping (HTRP) system where they received Low P (0.025 mM) or High P (0.25 mM) supply for 2 weeks. In hydroponics, root and shoot P and Zn concentrations were measured, root exudates were profiled using both Fourier-Transform-Infrared spectroscopy and gas-chromatography-mass spectrometry and previously published RNAseq data from roots was re-examined. In HTRP experiments, RSA (main and lateral root number and lateral root length) was assessed and the tissue-specific distribution of P was determined using micro-particle-induced-X-ray emission. The C6 accession had greater root and shoot biomass than the F103 accession, but the latter had a larger shoot P concentration than the C6 accession, regardless of the P supply in the hydroponic system. The F103 accession had a larger shoot Zn concentration than the C6 accession in the High P treatment. Although the F103 accession had a larger number of lateral roots, which were also longer than in the C6 accession, the C6 accession released a larger quantity and number of polar compounds than the F103 accession. A larger number of P-responsive genes were found in the Low P treatment in roots of the F103 accession than in roots of the C6 accession. Expression of genes linked with "phosphate starvation" was up-regulated, while those linked with iron homeostasis were down-regulated in the Low P treatment. CONCLUSIONS: The results illustrate large within-species variability in root acclimatory responses to P supply in the composition of root exudates, RSA and gene expression, but not in P distribution in root cross sections, enabling P sufficiency in the two B. oleracea accessions studied.

Arabidopsis halleri shows hyperbioindicator behaviour for Pb and leaf Pb accumulation spatially separated from Zn.

Lead (Pb) ranks among the most problematic environmental pollutants. Background contamination of soils is nearly ubiquitous, yet plant Pb accumulation is barely understood. In a survey covering 165 European populations of the metallophyte Arabidopsis halleri, several field samples had indicated Pb hyperaccumulation, offering a chance to dissect plant Pb accumulation. Accumulation of Pb was analysed in A. halleri individuals from contrasting habitats under controlled conditions to rule out aerial deposition as a source of apparent Pb accumulation. Several elemental imaging techniques were employed to study the spatial distribution and ligand environment of Pb. Regardless of genetic background, A. halleri individuals showed higher shoot Pb accumulation than A. thaliana. However, dose-response curves revealed indicator rather than hyperaccumulator behaviour. Xylem sap data and elemental imaging unequivocally demonstrated the in planta mobility of Pb. Highest Pb concentrations were found in epidermal and vascular tissues. Distribution of Pb was distinct from that of the hyperaccumulated metal zinc. Most Pb was bound by oxygen ligands in bidentate coordination. A. halleri accumulates Pb whenever soil conditions render Pb phytoavailable. Considerable Pb accumulation under such circumstances, even in leaves of A. thaliana, strongly suggests that Pb can enter food webs and may pose a food safety risk.

Phosphorus-zinc interactions in cotton: consequences for biomass production and nutrient-use efficiency in photosynthesis.

The fragmentary information on phosphorus (P) × zinc (Zn) interactions in plants warrants further study, particularly in plants known for their high P and Zn requirements, such as cotton (Gossypium hirsutum L.). The objective of this study was to investigate the effect of P × Zn interactions in a modern cultivar of cotton grown hydroponically. Biomass, mineral nutrition and photosynthetic parameters were monitored in plants receiving contrasting combinations of P and Zn supply. Root biomass, length and surface area were similar in plants with low P and/or low Zn supply to those in plants grown with high P and high Zn supply, reflecting an increased root/shoot biomass quotient when plants lack sufficient P or Zn for growth. Increasing P supply and reducing Zn supply increased shoot P concentrations, whilst shoot Zn concentrations were influenced largely by Zn supply. A balanced P × Zn supply (4 mM P × 4 µM Zn) enabled greatest biomass accumulation, while an imbalanced supply of these nutrients led to Zn deficiency, P toxicity or Zn toxicity. Net photosynthetic rate, stomatal conductance, transpiration rate and instantaneous carboxylation efficiency increased as P or Zn supply increased. Although increasing P supply reduced the P-use efficiency in photosynthesis (PUEP) and increasing Zn supply reduced the Zn-use efficiency in photosynthesis (ZnUEP), increasing Zn supply at a given P supply increased PUEP and increasing P supply at a given Zn supply increased ZnUEP. These results suggest that agricultural management strategies should seek for balanced mineral nutrition to optimize yields and resource-use efficiencies.

Spatially resolved analysis of variation in barley (Hordeum vulgare) grain micronutrient accumulation.

Genetic biofortification requires knowledge on natural variation and the underlying mechanisms of micronutrient accumulation. We therefore studied diversity in grain micronutrient concentrations and spatial distribution in barley (Hordeum vulgare), a genetically tractable model cereal and an important crop with widespread cultivation. We assembled a diverse collection of barley cultivars and landraces and analysed grain micronutrient profiles in genebank material and after three independent cultivations. Lines with contrasting grain zinc (Zn) accumulation were selected for in-depth analysis of micronutrient distribution within the grain by micro-proton-induced X-ray emission (µ-PIXE). Also, we addressed association with grain cadmium (Cd) accumulation. The analysis of > 120 lines revealed substantial variation, especially in grain Zn concentrations. A large fraction of this variation is due to genetic differences. Grain dissection and µ-PIXE analysis of contrasting lines showed that differences in grain Zn accumulation apply to all parts of the grain including the endosperm. Cd concentrations exceeded the Codex Alimentarius threshold in most of the representative barley lines after cultivation in a Cd-contaminated agricultural soil. Two important conclusions for biofortification are: first, high-Zn grains contain more Zn also in the consumed parts of the grain; and second, higher micronutrient concentrations are strongly associated with higher Cd accumulation.

Gomphrena claussenii, a novel metal-hypertolerant bioindicator species, sequesters cadmium, but not zinc, in vacuolar oxalate crystals.

Gomphrena claussenii is a recently described zinc (Zn)- and cadmium (Cd)-hypertolerant Amaranthaceae species displaying a metal bioindicator Zn/Cd accumulation response. We investigated the Zn and Cd distribution in stem and leaf tissues of G. claussenii at the cellular level, and determined metabolite profiles to investigate metabolite involvement in Zn and Cd sequestration. Gomphrena claussenii plants exposed to high Zn and Cd supply were analysed by scanning electron microscopy with energy-dispersive X-ray (SEM-EDX) and micro-proton-induced X-ray emission (micro-PIXE). In addition, gas chromatography-time of flight-mass spectrometry (GC-TOF-MS) was used to determine metabolite profiles on high Zn and Cd exposure. Stem and leaf tissues of G. claussenii plants exposed to control and high Cd conditions showed the abundant presence of calcium oxalate (CaOx) crystals, but on high Zn exposure, their abundance was strongly reduced. Ca and Cd co-localized to the CaOx crystals in Cd-exposed plants. Citrate, malate and oxalate levels were all higher in shoot tissues of metal-exposed plants, with oxalate levels induced 2.6-fold on Zn exposure and 6.4-fold on Cd exposure. Sequestration of Cd in vacuolar CaOx crystals of G. claussenii is found to be a novel mechanism to deal with Cd accumulation and tolerance.

De novo transcriptome assembly of hyperaccumulating Noccaea praecox for gene discovery.

Hyperaccumulators are a group of plant species that accumulate high concentrations of one or more metal(loid)s in their above-ground tissues without showing any signs of toxicity. Several hyperaccumulating species belong to the Brassicaceae family, among them the Cd and Zn hyperaccumulator Noccaea praecox. In this paper, we present de novo transcriptome assembled from two naturally occurring N. praecox populations growing in (i) metal-enriched soil and (ii) soil non-contaminated with metals (control site). Total RNA was extracted from the leaves of both populations. We obtained 801,935,101 reads, which were successfully assembled and annotated. The resulting assembly contains 135,323 transcripts, with 103,396 transcripts (76.4%) annotated with at least one function and encoding 53,142 putative proteins. Due to its close relationship with the hyperaccumulating model species N. cearulescens, it will be possible to derive protein functions from sequence comparisons with this species. Comparisons will highlight common and differing pathways of metal acquisition, storage, and detoxification which will allow us to expand our knowledge of these processes.

Gold-Assisted Molecular Imaging of Organic Tissue by MeV Secondary Ion Mass Spectrometry.

The quality of molecular imaging by means of MeV primary ion-induced secondary ion mass spectrometry by coating with gold was evaluated on different reference organic molecules and plant samples. The enhancement of the secondary ion yield was evident for the majority of the studied analytes, reaching the highest values at gold thicknesses between 0.5 and 2 nm, and increased the intensity up to 5-fold for reference samples and >2-fold for specific peaks within the plant sample. Improved propagation of the electric field due to the target potential on otherwise electrically insulating plant samples was also evident through improved image resolution and by reducing the background in mass spectra. However, detection of several molecules was significantly decreased at even at 1 nm thick gold layer. The results indicated that an optimized sequence of analysis is required to reliably interpret results.

Exploring the potential of cold plasma treatment followed by zinc-priming for biofortification of buckwheat sprouts.

Increasing the concentration of an element in edible produce (i.e., biofortification) can mitigate the element deficiency in humans. Sprouts are small but popular part of healthy diets providing vitamins and essential elements throughout the year. Element composition of sprouts can easily be amended, e.g., by soaking the grains in element-rich solution before germination (grain-priming). In addition, pre-treatment of grains to improve element translocation from the solution into the grain may further enhance the element concentration in the sprout. Cold plasma technique could provide such solution, as it increases wettability and water uptake of grains. Grains of common buckwheat (Fogopyrum esculentum Moench) were pre-treated/ untreated with cold plasma and soaked in ZnCl2 solution/pure water. Germination tests, α-amylase activity, grain hydrophilic properties and water uptake were assessed. Element composition of grain tissues and of sprouts was assessed by micro-particle-induced-X-ray emission and X-ray fluorescence spectroscopy, respectively. Grain-priming increased Zn concentration in shoots of common buckwheat sprouts more than five-times, namely from 79 to 423 mg Zn kg-1 dry weight. Cold plasma treatment increased grain wettability and water uptake into the grain. However, cold plasma pre-treatment followed by grain-priming with ZnCl2 did not increase Zn concentration in different grain tissues or in the sprouts more than the priming alone, but rather decreased the Zn concentration in sprout shoots (average ± standard error: 216 ± 6.13 and 174 ± 7.57 mg Zn kg-1 dry weight, respectively). When the fresh weight portion of whole sprouts (i.e., of roots and shoots) was considered, comparable average requirements of Zn, namely 24.5 % and 35 % for adult men and women would be satisfied by consuming cold plasma pre-treated and not pre-treated grains. Potential advantages of cold plasma pre-treatment need to be tested further, mainly to optimize the duration of soaking required to produce Zn-enriched sprouts.

Tissue-specific calcium and magnesium allocation to explain differences in bulk concentration in leaves of one-year-old seedlings of two olive (Olea europaea L.) cultivars.

Olive tree (Olea europaea L.) leaves have recently been recognised as a valuable source in cosmetic and pharmaceutical industry as well as in preparation of health-supporting beverages. Little is known about the element composition of olive leaves and almost nothing about tissue-specific allocation of elements. Element composition and tissue-specific distribution were determined in leaves of two olive cultivars, Leccino and Istarska bjelica using micro-particle induced X-ray emission (micro-PIXE). In leaves of the Istarska bjelica cultivar larger bulk concentrations of potassium, sodium, molybdenum and boron, but smaller concentrations of calcium and magnesium were found than in leaves of the Leccino cultivar. Tissue-specific investigation revealed that larger concentration of calcium in epidermis and in leaf blade tissues (secondary veins, palisade and spongy mesophyll) contributed to the larger leaf bulk calcium concentration in the Leccino cultivar. For magnesium, all leaf tissues, except the bundle sheath cells and consequently the main vascular bundle, contributed to the larger bulk concentration in the Leccino cultivar. Potassium was not predominant in any of the leaf tissues examined, while sodium and molybdenum were below the limit of detection, and boron not detectable by micro-PIXE. The results indicate that sinks for calcium and magnesium are stronger in specific leaf tissues of the Leccino than of the Istarska bjelica cultivar. The new understanding of tissue-specific allocation of elements in leaves of olive will serve as a basis for detailed studies into the effects of foliar and/or soil fertilisers in olive.

Imaging of Potassium and Calcium Distribution in Plant Tissues and Cells to Monitor Stress Response and Programmed Cell Death.

In plants, the response to stress, such as salinity, pathogen attack, drought, high concentration of metals, hyperthermia, and hypothermia, is usually accompanied by potassium ion (K+) leakage from the cytosol to the cell wall, mediated by plasma membrane cation conductivity. Stress-induced electrolyte leakage co-occurs with accumulation of reactive oxygen species (ROS) and calcium ions (Ca2+) and often results in programmed cell death (PCD). The development of X-ray and mass spectrometry (MS) based imaging techniques has enabled insight into the spatial tissue and cell-specific redistribution of major and trace elements during the stress response. In this chapter a workflow for sample preparation, imaging, and image analysis by X-ray and MS based techniques is presented.

Immobilization of chromium enhanced by arbuscular mycorrhizal fungi in semi-aquatic habitats with biochar addition.

Arbuscular mycorrhizal fungi (AMF) exhibit great potential in heavy-metal immobilization in semi-aquatic habitats. Under high heavy-metal stress, however, the role of AMF is limited, and the detoxification mechanism of AMF in heavy metals' stabilization remains unclear. This study investigated the effects of AMF on a wetland plant (Iris pseudacorus) and chromium (Cr) immobilization at different water depths in semi-aquatic habitats with biochar addition. Results showed that AMF increased the physiological and photosynthetic functions in I. pseudacorus under Cr exposures. Besides, AMF alleviated the accumulation of reactive oxygen species and lipid peroxidation by enhancing the antioxidant enzyme activities. AMF and biochar significantly decreased Cr concentrations in outlet water and increased Cr accumulation in I. pseudacorus. Besides, biochar also vastly improved Cr accumulation in the substrate under the fluctuating water depth. AMF reduced Cr bioavailability in the substrate, with Cr (Ⅵ) concentrations and acid-soluble forms of Cr decreased by 0.3-64.5% and 19.0-40.8%, respectively. Micro-proton-induced X-ray emission was used to determine element localization and revealed that AMF improved the nutrients uptake by wetland plants and inhibited Cr translocation from roots to shoots. Overall, this study demonstrated that the interaction between AMF and biochar could significantly enhance the immobilization of high Cr concentrations in semi-aquatic habitats.

The Effect of Feeding with Central European Local Mulberry Genotypes on the Development and Health Status of Silkworms and Quality Parameters of Raw Silk.

Silkworm rearing activities ceased in the 1970's in several European countries. Attempts on the re-establishment of ecological and sustainable sericulture in Slovenia and Hungary are ongoing. The aim of the study was to assess the usability of locally adapted mulberry genotypes for sericulture and to estimate connections between leaf compound and silkworm performance parameters. A controlled feeding experiment of silkworms was performed to test the influence of leaves from selected trees on the growth of larvae, the health and microbiological status of larvae (e.g., gut bacterial microbiome, Bombyx mori nucleopolyhedrovirus infection), weight of cocoons and raw silk parameters. The Slovenian and Hungarian mulberry genotypes had significantly higher total protein contents, and lower total phenolic contents and differed significantly in some individual phenolics compared to the reference sericultural and fruit varieties. Significant differences were found in the contents of the macro- and microelements, namely S, Mn, Fe, and Sr. Based on correlative statistics and multivariate analysis, a combined positive influence of proteins, specific phenolics, and microelements on larval growth and silk thread parameters was predicted. The results of the study indicate that selected local Slovenian and Hungarian mulberry varieties are suitable for high-quality silk cocoon and raw silk production.

The Halophyte Species Solanum chilense Dun. Maintains Its Reproduction despite Sodium Accumulation in Its Floral Organs.

Salinity is a growing global concern that affects the yield of crop species, including tomato (Solanum lycopersicum). Its wild relative Solanum chilense was reported to have halophyte properties. We compared salt resistance of both species during the reproductive phase, with a special focus on sodium localization in the flowers. Plants were exposed to NaCl from the seedling stage. Salinity decreased the number of inflorescences in both species but the number of flowers per inflorescence and sepal length only in S. lycopersicum. External salt supply decreased the stamen length in S. chilense, and it was associated with a decrease in pollen production and an increase in pollen viability. Although the fruit set was not affected by salinity, fruit weight and size decreased in S. lycopersicum. Concentrations and localization of Na, K, Mg, and Ca differed in reproductive structures of both species. Inflorescences and fruits of S. chilense accumulated more Na than S. lycopersicum. Sodium was mainly located in male floral organs of S. chilense but in non-reproductive floral organs in S. lycopersicum. The expression of Na transporter genes differed in flowers of both species. Overall, our results indicated that S. chilense was more salt-resistant than S. lycopersicum during the reproductive phase and that differences could be partly related to dissimilarities in element distribution and transport in flowers.

New insights into globoids of protein storage vacuoles in wheat aleurone using synchrotron soft X-ray microscopy.

Mature developed seeds are physiologically and biochemically committed to store nutrients, principally as starch, protein, oils, and minerals. The composition and distribution of elements inside the aleurone cell layer reflect their biogenesis, structural characteristics, and physiological functions. It is therefore of primary importance to understand the mechanisms underlying metal ion accumulation, distribution, storage, and bioavailability in aleurone subcellular organelles for seed fortification purposes. Synchrotron radiation soft X-ray full-field imaging mode (FFIM) and low-energy X-ray fluorescence (LEXRF) spectromicroscopy were applied to characterize major structural features and the subcellular distribution of physiologically important elements (Zn, Fe, Na, Mg, Al, Si, and P). These direct imaging methods reveal the accumulation patterns between the apoplast and symplast, and highlight the importance of globoids with phytic acid mineral salts and walls as preferential storage structures. C, N, and O chemical topographies are directly linked to the structural backbone of plant substructures. Zn, Fe, Na, Mg, Al, and P were linked to globoid structures within protein storage vacuoles with variable levels of co-localization. Si distribution was atypical, being contained in the aleurone apoplast and symplast, supporting a physiological role for Si in addition to its structural function. These results reveal that the immobilization of metals within the observed endomembrane structures presents a structural and functional barrier and affects bioavailability. The combination of high spatial and chemical X-ray microscopy techniques highlights how in situ analysis can yield new insights into the complexity of the wheat aleurone layer, whose precise biochemical composition, morphology, and structural characteristics are still not unequivocally resolved.

Localization of aluminium in tea (Camellia sinensis) leaves using low energy X-ray fluorescence spectro-microscopy.

Information on localization of Al in tea leaf tissues is required in order to better understand Al tolerance mechanism in this Al-accumulating plant species. Here, we have used low-energy X-ray fluorescence spectro-microscopy (LEXRF) to study localization of Al and other low Z-elements, namely C, O, Mg, Si and P, in fully developed leaves of the tea plant [Camellia sinensis (L.) O. Kuntze]. Plants were grown from seeds for 3 months in a hydroponic solution, and then exposed to 200 microM AlCl(3) for 2 weeks. Epidermal-mesophyll and xylem phloem regions of 20 microm thick cryo-fixed freeze-dried tea-leaf cross-sections were raster scanned with 1.7 and 2.2 keV excitation energies to reach the Al-K and P-K absorption edges. Al was mainly localized in the cell walls of the leaf epidermal cells, while almost no Al signal was obtained from the leaf symplast. The results suggest that the retention of Al in epidermal leaf apoplast represent the main tolerance mechanism to Al in tea plants. In addition LEXRF proved to be a powerful tool for localization of Al in plant tissues, which can help in our understanding of the processes of Al uptake, transport and tolerance in plants.

Unravelling homeostasis effects of phosphorus and zinc nutrition by leaf photochemistry and metabolic adjustment in cotton plants.

Phosphorus (P) and zinc (Zn) uptake and its physiological use in plants are interconnected and are tightly controlled. However, there is still conflicting information about the interactions of these two nutrients, thus a better understanding of nutritional homeostasis is needed. The objective of this work was to evaluate responses of photosynthesis parameters, P-Zn nutritional homeostasis and antioxidant metabolism to variation in the P × Zn supply of cotton (Gossypium hirsutum L.). Plants were grown in pots and watered with nutrient solution containing combinations of P and Zn supply. An excess of either P or Zn limited plant growth, reduced photosynthesis-related parameters, and antioxidant scavenging enzymes. Phosphorus uptake favoured photochemical dissipation of energy decreasing oxidative stress, notably on Zn-well-nourished plants. On the other hand, excessive P uptake reduces Zn-shoot concentration and decreasing carbonic anhydrase activity. Adequate Zn supply facilitated adaptation responses to P deficiency, upregulating acid phosphatase activity, whereas Zn and P excess were alleviated by increasing P and Zn supply, respectively. Collectively, the results showed that inter ionic effects of P and Zn uptake affected light use and CO2 assimilation rate on photosynthesis, activation of antioxidant metabolism, acid phosphatase and carbonic anhydrase activities, and plant growth-related responses to different extents.

Mineral Element Composition in Grain of Awned and Awnletted Wheat (Triticum aestivum L.) Cultivars: Tissue-Specific Iron Speciation and Phytate and Non-Phytate Ligand Ratio.

In wheat (Triticum aestivum L.), the awns-the bristle-like structures extending from lemmas-are photosynthetically active. Compared to awned cultivars, awnletted cultivars produce more grains per unit area and per spike, resulting in significant reduction in grain size, but their mineral element composition remains unstudied. Nine awned and 11 awnletted cultivars were grown simultaneously in the field. With no difference in 1000-grain weight, a larger calcium and manganese-but smaller iron (Fe) concentrations-were found in whole grain of awned than in awnletted cultivars. Micro X-ray absorption near edge structure analysis of different tissues of frozen-hydrated grain cross-sections revealed that differences in total Fe concentration were not accompanied by differences in Fe speciation (64% of Fe existed as ferric and 36% as ferrous species) or Fe ligands (53% were phytate and 47% were non-phytate ligands). In contrast, there was a distinct tissue-specificity with pericarp containing the largest proportion (86%) of ferric species and nucellar projection (49%) the smallest. Phytate ligand was predominant in aleurone, scutellum and embryo (72%, 70%, and 56%, respectively), while nucellar projection and pericarp contained only non-phytate ligands. Assuming Fe bioavailability depends on Fe ligands, we conclude that Fe bioavailability from wheat grain is tissue specific.

Breeding Buckwheat for Increased Levels of Rutin, Quercetin and Other Bioactive Compounds with Potential Antiviral Effects.

Common buckwheat (Fagopyrum esculentum Moench) and Tartary buckwheat (Fagopyrum tataricum (L.) Gaertn.) are sources of many bioactive compounds, such as rutin, quercetin, emodin, fagopyrin and other (poly)phenolics. In damaged or milled grain under wet conditions, most of the rutin in common and Tartary buckwheat is degraded to quercetin by rutin-degrading enzymes (e.g., rutinosidase). From Tartary buckwheat varieties with low rutinosidase activity it is possible to prepare foods with high levels of rutin, with the preserved initial levels in the grain. The quercetin from rutin degradation in Tartary buckwheat grain is responsible in part for inhibition of α-glucosidase in the intestine, which helps to maintain normal glucose levels in the blood. Rutin and emodin have the potential for antiviral effects. Grain embryos are rich in rutin, so breeding buckwheat with the aim of producing larger embryos may be a promising strategy to increase the levels of rutin in common and Tartary buckwheat grain, and hence to improve its nutritional value.