The Effect of Exogenous Cadmium and Zinc Applications on Cadmium, Zinc and Essential Mineral Bioaccessibility in Three Lines of Rice That Differ in Grain Cadmium Accumulation.

Millions of people around the world rely on rice (Oryza sativa) for a significant portion of daily calories, but rice is a relatively poor source of essential micronutrients like iron and zinc. Rice has been shown to accumulate alarmingly high concentrations of toxic elements, such as cadmium. Cadmium in foods can lead to renal failure, bone mineral density loss, cancer, and significant neurotoxicological effects. Several strategies to limit cadmium and increase micronutrient density in staple food crops like rice have been explored, but even when cadmium concentrations are reduced by a management strategy, total cadmium levels in rice grain are an unreliable means of estimating human health risk because only a fraction of the minerals in grains are bioaccessible. The goal of this work was to assess the influence of cadmium and zinc supplied to plant roots on the bioaccessibility of cadmium and essential minerals from grains of three rice lines (GSOR 310546/low grain Cd, GSOR 311667/medium grain Cd, and GSOR 310428/high grain Cd) that differed in grain cadmium accumulation. Treatments consisted of 0 µM Cd + 2 µM Zn (c0z2), 1 µM Cd + 2 µM Zn (c1z2), or 1 µM Cd + 10 µM Zn (c1z10). Our results revealed that an increased grain cadmium concentration does not always correlate with increased cadmium bioaccessibility. Among the three rice lines tested, Cd bioaccessibility increased from 2.5% in grains from the c1z2 treatment to 17.7% in grains from the c1z10 treatment. Furthermore, Cd bioccessibility in the low-Cd-accumulating line was significantly higher than the high line in c1z10 treatment. Zinc bioaccessibility increased in the high-cadmium-accumulating line when cadmium was elevated in grains, and in the low-cadmium line when both cadmium and zinc were increased in the rice grains. Our results showed that both exogenous cadmium and elevated zinc treatments increased the bioaccessibility of other minerals from grains of the low- or high-grain cadmium lines of rice. Differences in mineral bioaccessibility were dependent on rice line. Calculations also showed that increased cadmium bioaccessibility correlated with increased risk of dietary exposure to consumers. Furthermore, our results suggest that zinc fertilization increased dietary exposure to cadmium in both high and low lines. This information can inform future experiments to analyze genotypic effects of mineral bioavailability from rice, with the goal of reducing cadmium absorption while simultaneously increasing zinc absorption from rice grains.

Seedborne Cercospora beticola Can Initiate Cercospora Leaf Spot from Sugar Beet (Beta vulgaris) Fruit Tissue.

Cercospora leaf spot (CLS) is a globally important disease of sugar beet (Beta vulgaris) caused by the fungus Cercospora beticola. Long-distance movement of C. beticola has been indirectly evidenced in recent population genetic studies, suggesting potential dispersal via seed. Commercial sugar beet "seed" consists of the reproductive fruit (true seed surrounded by maternal pericarp tissue) coated in artificial pellet material. In this study, we confirmed the presence of viable C. beticola in sugar beet fruit for 10 of 37 tested seed lots. All isolates harbored the G143A mutation associated with quinone outside inhibitor resistance, and 32 of 38 isolates had reduced demethylation inhibitor sensitivity (EC50 > 1 µg/ml). Planting of commercial sugar beet seed demonstrated the ability of seedborne inoculum to initiate CLS in sugar beet. C. beticola DNA was detected in DNA isolated from xylem sap, suggesting the vascular system is used to systemically colonize the host. We established nuclear ribosomal internal transcribed spacer region amplicon sequencing using the MinION platform to detect fungi in sugar beet fruit. Fungal sequences from 19 different genera were identified from 11 different sugar beet seed lots, but Fusarium, Alternaria, and Cercospora were consistently the three most dominant taxa, comprising an average of 93% relative read abundance over 11 seed lots. We also present evidence that C. beticola resides in the pericarp of sugar beet fruit rather than the true seed. The presence of seedborne inoculum should be considered when implementing integrated disease management strategies for CLS of sugar beet in the future.

Cultivar Differences in the Biochemical and Physiological Responses of Common Beans to Aluminum Stress.

Soil conditions leading to high levels of available aluminum are detrimental to plant growth, but data are limited on genotypic differences in tolerance to aluminum stress in some crops. The aim of this study was to examine the morphological, biochemical, and physiological changes in roots and shoots of 25 common bean (Phaseolus vulgaris L.) cultivars (Pinto market class) under aluminum (Al) treatment. Additionally, this study aimed to assess the range of responses amongst the common bean cultivars relative to their Al toxicity tolerance and sensitivity. Plants were grown hydroponically using a simplified nutrient solution with or without 20 µM AlCl3. Reactive oxygen species (ROS), activities of the antioxidant enzymes superoxide dismutase (SOD) and guaiacol peroxidase (POD), and malondialdehyde (MDA) concentration, an indicator of lipid peroxidation, were measured to establish the effects of Al treatment on the plants. In addition, growth parameters such as shoot and root dry weight, root-to-shoot ratio, root elongation, and root volume changes were also investigated. The cultivar effect was significant for all the measured parameters, except for shoot dry weight. Inhibition of the root and shoot dry weight for selected common bean cultivars shows that the response of common bean to Al stress is genotype-specific. Additionally, Al-induced root elongation inhibition and root volume changes varied among the cultivars. Most cultivars had significantly higher SOD activity (20 of 25 cultivars) and POD activity (12 cultivars) under AlCl3 treatment compared to the controls. A positive significant correlation was observed between MDA and ROS, showing that Al stress induced the accumulation of ROS along with an increase in lipid peroxidation. According to the results of this study, Arapaho and AC Island cultivars could potentially be used in the future production of common beans under Al stress. Therefore, these two cultivars could also be included in Al tolerance breeding programs.

Stacking disease resistance and mineral biofortification in cassava varieties to enhance yields and consumer health.

Delivering the benefits of agricultural biotechnology to smallholder farmers requires that resources be directed towards staple food crops. To achieve effect at scale, beneficial traits must be integrated into multiple, elite farmer-preferred varieties with relevance across geographical regions. The staple root crop cassava (Manihot esculenta) is consumed for dietary calories by more than 800 million people, but its tuberous roots provide insufficient iron and zinc to meet nutritional needs. In Africa, cassava yields are furthermore limited by the virus diseases, cassava mosaic disease (CMD) and cassava brown streak disease (CBSD). In this study, we strove to develop cassava displaying high-level resistance to CBSD and CMD to attain food and economic security for cassava farmers, along with biofortified levels of iron and zinc to enhance consumer health. RNAi-mediated technology was used to achieve resistance to CBSD in two East African and one Nigerian farmer-preferred cultivars that harboured resistance to CMD. The Nigerian cvs. TMS 95/0505 and TMS 91/02324 were modified with T-DNA imparting resistance to CBSD, along with AtIRT1 (major iron transporter) and AtFER1 (ferritin) transgenes to achieve nutritionally significant levels of iron and zinc in cassava storage roots (145 and 40 µg/g dry weight, respectively). The inherent resistance to CMD was maintained in all four disease resistant and mineral enhanced cassava cultivars described here, demonstrating that this technique could be deployed across multiple farmer-preferred varieties to benefit the food and nutritional security of consumers in Africa.

Colocalization of sucrose synthase expression and sucrose storage in the sugarbeet taproot indicates a potential role for sucrose catabolism in sucrose accumulation.

Sucrose metabolism is believed to have a central role in promoting sink strength and sucrose storage in the sugarbeet taproot. How sucrose accumulation is increased by sucrose-degrading enzymes, however, is a paradox. To elucidate roles for sucrose-degrading activities in sucrose accumulation, relationships between the intercellular location of sucrose-catabolizing enzymes and sites of sucrose accumulation were determined in the sugarbeet taproot. Sucrose storage was evident in parenchyma cells of the outer cortex, rays, and rings of parenchyma tissue, but was absent in phloem, the vascular cambium, cells surrounding these tissues, or cells surrounding xylem. Sucrose synthase, which was primarily responsible for sucrose catabolism throughout the taproot, was expressed in similar cell and tissue types to those accumulating sucrose. Colocalization of sucrose synthase with sucrose accumulation, as well as sucrose synthase localization near the tonoplast, suggests a role for the enzyme in generating metabolic energy to fuel sucrose sequestration in the vacuole. Localization near the plasma membrane also suggests a role for sucrose synthase in supplying substrates for cell wall biosynthesis. By utilizing sucrose for ATP or cell wall biosynthesis, sucrose synthase likely maintains the source-to-sink sucrose gradient that drives sucrose transport into the root, thereby promoting sugarbeet root sink strength.

Author Correction: Biofortification of field-grown cassava by engineering expression of an iron transporter and ferritin.

In the version of this article initially published, a relevant work was not cited. The following sentence has been inserted following the sentence ending "Aspergillus phytase" in the third paragraph of the article: "Overexpression of AtIRT1, AtNAS1 and bean FERRITIN in rice resulted in 3.8-fold higher iron and 1.8-fold higher zinc concentrations than in the wild-type control12." A corresponding reference has been added: 12. Boonyaves, K., Wu, T. Y., Gruissem, W. & Bhullar, N. K. Enhanced grain iron levels in rice expressing an IRON-REGULATED METAL TRANSPORTER, NICOTIANAMINE SYNTHASE, and FERRITIN gene cassette. Front. Plant Sci. 8, 130 (2017). The error has been corrected in the HTML and PDF versions of the article.

Atorvastatin Decreases Renal Menaquinone-4 Formation in C57BL/6 Male Mice.

BACKGROUND: Menaquinone-4 (MK4), a vitamin K metabolite, is converted from phylloquinone through a process that requires intermediates of endogenous cholesterol production. Recent evidence suggests that MK4 is involved in kidney function. OBJECTIVE: The purpose of this study was to determine the effect of atorvastatin treatment on MK4 formation in young and old male mice. METHODS: C57BL/6 male mice (4-mo-old and 20-mo-old) were randomly assigned to either a diet containing 300 mg atorvastatin/kg with 3 mg phylloquinone/kg or a control diet containing 3 mg phylloquinone/kg for 8 wk. During week 8, all mice received deuterium-labeled phylloquinone in the diet. Labeled and unlabeled phylloquinone and MK4 in liver, kidney, brain, and intestine were measured by atmospheric pressure chemical ionization LC/MS. 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase gene expression was quantified by reverse transcriptase-PCR. Tissue MK4 and phylloquinone concentrations were compared between atorvastatin treatment groups with use of general linear models. RESULTS: There was no age-treatment interaction on MK4 tissue concentrations. In atorvastatin-treated mice, total MK4 and percentage of deuterium-labeled MK4 in kidney were both approximately 45% lower compared to values in mice not given atorvastatin (all P < 0.05). MK4 concentrations did not differ between groups in any other tissue measured. CONCLUSION: In male mice, atorvastatin reduced endogenous MK4 formation in the kidney, but not other organs. These observations are consistent with our hypothesis that cholesterol metabolism is involved in the generation of MK4. Further research is needed to understand potential regulatory mechanisms and the unique functions of MK4 in the kidney.

Biofortification of field-grown cassava by engineering expression of an iron transporter and ferritin.

Less than 10% of the estimated average requirement (EAR) for iron and zinc is provided by consumption of storage roots of the staple crop cassava (Manihot esculenta Crantz) in West African human populations. We used genetic engineering to improve mineral micronutrient concentrations in cassava. Overexpression of the Arabidopsis thaliana vacuolar iron transporter VIT1 in cassava accumulated three- to seven-times-higher levels of iron in transgenic storage roots than nontransgenic controls in confined field trials in Puerto Rico. Plants engineered to coexpress a mutated A. thaliana iron transporter (IRT1) and A. thaliana ferritin (FER1) accumulated iron levels 7-18 times higher and zinc levels 3-10 times higher than those in nontransgenic controls in the field. Growth parameters and storage-root yields were unaffected by transgenic fortification in our field data. Measures of retention and bioaccessibility of iron and zinc in processed transgenic cassava indicated that IRT1 + FER1 plants could provide 40-50% of the EAR for iron and 60-70% of the EAR for zinc in 1- to 6-year-old children and nonlactating, nonpregnant West African women.

Plasma Response to Deuterium-Labeled Vitamin K Intake Varies by TG Response, but Not Age or Vitamin K Status, in Older and Younger Adults.

Background: Phylloquinone is the primary form of vitamin K in the diet and circulation. Large intra- and interindividual variances in circulating phylloquinone have been partially attributed to age. However, little is known about the nondietary factors that influence phylloquinone absorption and metabolism. Similarly, it is not known if phylloquinone absorption is altered by the individual's existing vitamin K status. Objective: The purpose of this secondary substudy was to compare plasma response with deuterium-labeled phylloquinone intake in older and younger adults after dietary phylloquinone depletion and repletion. Methods: Forty-two older [mean ± SD age: 67.2 ± 8.0 y; body mass index (BMI; in kg/m2): 25.4 ± 4.6; n = 12 men, 9 women] and younger (mean ± SEM age: 31.8 ± 6.6 y; BMI: 25.5 ± 3.3; n = 9 men, 12 women) adults were maintained on sequential 28-d phylloquinone depletion (∼10 µg phylloquinone/d) and 28-d phylloquinone repletion (∼500 µg phylloquinone/d) diets. On the 23rd d of each diet phase, participants consumed deuterated phylloquinone-rich collard greens (2H-phylloquinone). Plasma and urinary outcome measures over 72 h were compared by age group, sex, and dietary phase via 2-factor repeated-measures ANOVA. Results: The plasma 2H-phylloquinone area under the curve (AUC) did not differ in response to phylloquinone depletion or repletion, but was 34% higher in older than in younger adults (P = 0.02). However, plasma 2H-phylloquinone AUC was highly correlated with the serum triglyceride (TG) AUC (r2 = 0.45). After adjustment for serum TG response, the age effect on the plasma 2H-phylloquinone AUC was no longer significant. Conclusions: Plasma 2H-phylloquinone response did not differ between phylloquinone depletion and repletion in older and younger adults. The age effect observed was explained by the serum TG response and was completely attenuated after adjustment. Plasma response to phylloquinone intake, therefore, seems to be a predominantly lipid-driven effect and not dependent on existing vitamin K status. More research is required to differentiate the effect of endogenous compared with exogenous lipids on phylloquinone absorption. This trial was registered at clinicaltrials.gov as NCT00336232.

Influence of alternative soil amendments on mycorrhizal fungi and cowpea production.

Alternative soil amendments (worm compost, pyrolyzed carbon [biochar]) and crop symbioses with arbuscular mycorrhizal (AM) fungi have the potential to reduce food production costs while promoting sustainable agriculture by improving soil quality and reducing commercial (N and P) fertilizer use. Our greenhouse studies investigated the influence of alternative soil amendments on AM fungi associated with cowpea (Vigna unguiculata [L.] Walp.) and common bean (Phaseolus vulgaris L.) by examining productivity and plant nutrition. We conducted an experiment to select a cowpea or common bean genotype based on AM fungal colonization, seed production, and seed nutritional content. We then grew the selected cowpea genotype (Resina) in low-fertility soil with 10 different soil amendments (combinations of biochar, worm compost, and/or commercial fertilizers) plus a non-amended control. There were no significant differences in AM fungal colonization of cowpea plants grow with different soil amendments. However, an amendment blend containing worm compost, biochar, and 50% of the typically recommended commercial fertilizer rate produced plants with similar aboveground biomass, protein concentration, and total protein production, with increased tissue K, P, and Zn concentration and total content, compared to plants receiving only the recommended (100%) rate of commercial fertilizer. As previous research links uptake of P and Zn with plant-mycorrhizal symbioses, our results indicate cowpea nutritional benefits may be derived from AM partnership and alternative soil amendments. These synergies between alternative soil amendments and AM fungi may help reduce farm costs while maintaining or improving crop yield and nutrition, thus increasing global food and nutrition security.

In silico mapping of quantitative trait loci (QTL) regulating the milk ionome in mice identifies a milk iron locus on chromosome 1.

The breast-feeding neonate depends on mother's milk for both macronutrients and micronutrients including minerals. The goals of the present study were to document the effects of genetic background in mice on milk concentrations of select minerals and to use genome-wide association study (GWAS) to identify quantitative trait loci (QTL) regulating milk mineral concentrations. Milk samples from lactating mice in each of 31 different inbred strains of the mouse diversity panel (MDP) were analyzed by inductively coupled plasma-optical emission spectroscopy to determine the concentrations of calcium (Ca), copper (Cu), iron (Fe), potassium (K), magnesium (Mg), sodium (Na), phosphorus (P), sulfur (S), and zinc (Zn). GWAS identified a single pleiotropic milk mineral concentration QTL (Mmcq) on chromosome 3 for Ca, Mg, and P. For the remaining minerals, six QTL were detected for Fe, four for K, three for Zn, and one for S. Intersecting the Mmcq with published chromatin immunoprecipitation sequence data identified 15 out of 4633 high-linkage disequilibrium single-nucleotide polymorphisms that resided in signal transducer and activation of transcription 5 (STAT5) binding regions. A milk Fe-associated locus (Mmcq9) on chromosome 1 contained an SNP that localized to a STAT5 binding region and intersected with a HOMER motif predicted to bind the transcriptional regulator E74-Like ETS transcription factor 5. This locus also contained the genes for solute carrier family (Slc) members Slc9a2, Slc9a4, Slc39a10, and Slc40a1. Expression analysis of these transporters supports the conclusion that Slc9a2 and Slc40a1 within the mammary gland could mediate the effect of Mmcq9 on milk Fe concentration.

Genetic diversity and association mapping of mineral element concentrations in spinach leaves.

BACKGROUND: Spinach is a useful source of dietary vitamins and mineral elements. Breeding new spinach cultivars with high nutritional value is one of the main goals in spinach breeding programs worldwide, and identification of single nucleotide polymorphism (SNP) markers for mineral element concentrations is necessary to support spinach molecular breeding. The purpose of this study was to conduct a genome-wide association study (GWAS) and to identify SNP markers associated with mineral elements in the USDA-GRIN spinach germplasm collection. RESULTS: A total of 14 mineral elements: boron (B), calcium (Ca), cobalt (Co), copper (Cu), iron (Fe), potassium (K), magnesium (Mg), manganese (Mn), molybdenum (Mo), sodium (Na), nickel (Ni), phosphorus (P), sulfur (S), and zinc (Zn) were evaluated in 292 spinach accessions originally collected from 29 countries. Significant genetic variations were found among the tested genotypes as evidenced by the 2 to 42 times difference in mineral concentrations. A total of 2402 SNPs identified from genotyping by sequencing (GBS) approach were used for genetic diversity and GWAS. Six statistical methods were used for association analysis. Forty-five SNP markers were identified to be strongly associated with the concentrations of 13 mineral elements. Only two weakly associated SNP markers were associated with K concentration. Co-localized SNPs for different elemental concentrations were discovered in this research. Three SNP markers, AYZV02017731_40, AYZV02094133_57, and AYZV02281036_185 were identified to be associated with concentrations of four mineral components, Co, Mn, S, and Zn. There is a high validating correlation coefficient with r > 0.7 among concentrations of the four elements. Thirty-one spinach accessions, which rank in the top three highest concentrations in each of the 14 mineral elements, were identified as potential parents for spinach breeding programs in the future. CONCLUSIONS: The 45 SNP markers strongly associated with the concentrations of the 13 mineral elements: B, Ca, Co, Cu, Fe, Mg, Mn, Mo, Na, Ni, P, S, and Zn could be used in breeding programs to improve the nutritional quality of spinach through marker-assisted selection (MAS). The 31 spinach accessions with high concentrations of one to several mineral elements can be used as potential parents for spinach breeding programs.

Use of a "Super-child" Approach to Assess the Vitamin A Equivalence of Moringa oleifera Leaves, Develop a Compartmental Model for Vitamin A Kinetics, and Estimate Vitamin A Total Body Stores in Young Mexican Children.

Background: Worldwide, an estimated 250 million children <5 y old are vitamin A (VA) deficient. In Mexico, despite ongoing efforts to reduce VA deficiency, it remains an important public health problem; thus, food-based interventions that increase the availability and consumption of provitamin A-rich foods should be considered.Objective: The objectives were to assess the VA equivalence of 2H-labeled Moringa oleifera (MO) leaves and to estimate both total body stores (TBS) of VA and plasma retinol kinetics in young Mexican children.Methods: ß-Carotene was intrinsically labeled by growing MO plants in a 2H2O nutrient solution. Fifteen well-nourished children (17-35 mo old) consumed puréed MO leaves (1 mg ß-carotene) and a reference dose of [13C10]retinyl acetate (1 mg) in oil. Blood (2 samples/child) was collected 10 times (2 or 3 children each time) over 35 d. The bioefficacy of MO leaves was calculated from areas under the composite "super-child" plasma isotope response curves, and MO VA equivalence was estimated through the use of these values; a compartmental model was developed to predict VA TBS and retinol kinetics through the use of composite plasma [13C10]retinol data. TBS were also estimated with isotope dilution.Results: The relative bioefficacy of ß-carotene retinol activity equivalents from MO was 28%; VA equivalence was 3.3:1 by weight (0.56 µmol retinol:1 µmol ß-carotene). Kinetics of plasma retinol indicate more rapid plasma appearance and turnover and more extensive recycling in these children than are observed in adults. Model-predicted mean TBS (823 µmol) was similar to values predicted using a retinol isotope dilution equation applied to data from 3 to 6 d after dosing (mean ± SD: 832 ± 176 µmol; n = 7).Conclusions: The super-child approach can be used to estimate population carotenoid bioefficacy and VA equivalence, VA status, and parameters of retinol metabolism from a composite data set. Our results provide initial estimates of retinol kinetics in well-nourished young children with adequate VA stores and demonstrate that MO leaves may be an important source of VA.

Arabidopsis Glutaredoxin S17 Contributes to Vegetative Growth, Mineral Accumulation, and Redox Balance during Iron Deficiency.

Iron (Fe) is an essential mineral nutrient and a metal cofactor required for many proteins and enzymes involved in the processes of DNA synthesis, respiration, and photosynthesis. Iron limitation can have detrimental effects on plant growth and development. Such effects are mediated, at least in part, through the generation of reactive oxygen species (ROS). Thus, plants have evolved a complex regulatory network to respond to conditions of iron limitations. However, the mechanisms that couple iron deficiency and oxidative stress responses are not fully understood. Here, we report the discovery that an Arabidopsis thaliana monothiol glutaredoxin S17 (AtGRXS17) plays a critical role in the plants ability to respond to iron deficiency stress and maintain redox homeostasis. In a yeast expression assay, AtGRXS17 was able to suppress the iron accumulation in yeast ScGrx3/ScGrx4 mutant cells. Genetic analysis indicated that plants with reduced AtGRXS17 expression were hypersensitive to iron deficiency and showed increased iron concentrations in mature seeds. Disruption of AtGRXS17 caused plant sensitivity to exogenous oxidants and increased ROS production under iron deficiency. Addition of reduced glutathione rescued the growth and alleviates the sensitivity of atgrxs17 mutants to iron deficiency. These findings suggest AtGRXS17 helps integrate redox homeostasis and iron deficiency responses.

Genome-wide SNP identification, linkage map construction and QTL mapping for seed mineral concentrations and contents in pea (Pisum sativum L.).

BACKGROUND: Marker-assisted breeding is now routinely used in major crops to facilitate more efficient cultivar improvement. This has been significantly enabled by the use of next-generation sequencing technology to identify loci and markers associated with traits of interest. While rich in a range of nutritional components, such as protein, mineral nutrients, carbohydrates and several vitamins, pea (Pisum sativum L.), one of the oldest domesticated crops in the world, remains behind many other crops in the availability of genomic and genetic resources. To further improve mineral nutrient levels in pea seeds requires the development of genome-wide tools. The objectives of this research were to develop these tools by: identifying genome-wide single nucleotide polymorphisms (SNPs) using genotyping by sequencing (GBS); constructing a high-density linkage map and comparative maps with other legumes, and identifying quantitative trait loci (QTL) for levels of boron, calcium, iron, potassium, magnesium, manganese, molybdenum, phosphorous, sulfur, and zinc in the seed, as well as for seed weight. RESULTS: In this study, 1609 high quality SNPs were found to be polymorphic between 'Kiflica' and 'Aragorn', two parents of an F6-derived recombinant inbred line (RIL) population. Mapping 1683 markers including 75 previously published markers and 1608 SNPs developed from the present study generated a linkage map of size 1310.1 cM. Comparative mapping with other legumes demonstrated that the highest level of synteny was observed between pea and the genome of Medicago truncatula. QTL analysis of the RIL population across two locations revealed at least one QTL for each of the mineral nutrient traits. In total, 46 seed mineral concentration QTLs, 37 seed mineral content QTLs, and 6 seed weight QTLs were discovered. The QTLs explained from 2.4% to 43.3% of the phenotypic variance. CONCLUSION: The genome-wide SNPs and the genetic linkage map developed in this study permitted QTL identification for pea seed mineral nutrients that will serve as important resources to enable marker-assisted selection (MAS) for nutritional quality traits in pea breeding programs.

A rapid and efficient method to study the function of crop plant transporters in Arabidopsis.

Iron (Fe) is an essential micronutrient for humans. Fe deficiency disease is widespread and has led to extensive studies on the mechanisms of Fe uptake and storage, especially in staple food crops such as rice. However, studies of functionally related genes in rice and other crops are often time and space demanding. Here, we demonstrate that transgenic Arabidopsis suspension culture cells and Arabidopsis plants can be used as an efficient expression system for gain-of-function study of selected transporters, using Fe transporters as a proof-of-principle. The vacuolar membrane transporters OsVIT1 and OsVIT2 have been described to be important for iron sequestration, and disruption of these two genes leads to Fe accumulation in rice seeds. In this study, we have taken advantage of the fluorescent-tagged protein GFP-OsVIT1, which functionally complements the Fe hypersensitivity of ccc1 yeast mutant, to generate transgenic Arabidopsis suspension cell lines and plants. GFP-OsVIT1 was shown to localize on the vacuolar membrane using confocal microscopy and immunogold EM. More importantly, the Fe concentration, as well as the concentration of Zn, in the transgenic cell lines and plants were significantly increased compared to that in the WT. Taken together, our study shows that the heterologous expression of rice vacuolar membrane transporter OsVIT1 in Arabidopsis system is functional and effectively enhances iron accumulation, indicating an useful approach for studying other putative transporters of crop plants in this system.

Demonstrating a Nutritional Advantage to the Fast-Cooking Dry Bean (Phaseolus vulgaris L.).

Dry beans (Phaseolus vulgaris L.) are a nutrient-dense food rich in protein and micronutrients. Despite their nutritional benefits, long cooking times limit the consumption of dry beans worldwide, especially in nations where fuelwood for cooking is often expensive or scarce. This study evaluated the nutritive value of 12 dry edible bean lines that vary for cooking time (20-89 min) from four market classes (yellow, cranberry, light red kidney, and red mottled) of economic importance in bean-consuming regions of Africa and the Americas. When compared to their slower cooking counterparts within each market class, fast-cooking dry beans retain more protein and minerals while maintaining similar starch and fiber densities when fully cooked. For example, some of the highest protein and mineral retention values were measured in the fast-cooking yellow bean cultivar Cebo Cela, which offered 20% more protein, 10% more iron, and 10% more zinc with each serving when compared with Canario, a slow-cooking yellow bean that requires twice the cooking time to become palatable. A Caco-2 cell culture model also revealed the bioavailability of iron is significantly higher in faster cooking entries (r = -0.537, P = 0.009) as compared to slower cooking entries in the same market class. These findings suggest that fast-cooking bean varieties have improved nutritive value through greater nutrient retention and improved iron bioavailability.

Leaf Protein and Mineral Concentrations across the "Miracle Tree" Genus Moringa.

The moringa tree Moringa oleifera is a fast-growing, drought-resistant tree cultivated across the lowland dry tropics worldwide for its nutritious leaves. Despite its nutritious reputation, there has been no systematic survey of the variation in leaf nutritional quality across M. oleifera grown worldwide, or of the other species of the genus. To guide informed use of moringa, we surveyed protein, macro-, and micro- nutrients across 67 common garden samples of 12 Moringa taxa, including 23 samples of M. oleifera. Moringa oleifera, M. concanensis, M. stenopetala, an M. concanensis X oleifera hybrid, and M. longituba were highest in protein, with M. ruspoliana having the highest calcium levels. A protein-dry leaf mass tradeoff may preclude certain breeding possibilities, e.g. maximally high protein with large leaflets. These findings identify clear priorities and limitations for improved moringa varieties with traits such as high protein, calcium, or ease of preparation.

Effects of Fe deficiency on the protein profiles and lignin composition of stem tissues from Medicago truncatula in absence or presence of calcium carbonate.

UNLABELLED: Iron deficiency is a yield-limiting factor with major implications for crop production, especially in soils with high CaCO3. Because stems are essential for the delivery of nutrients to the shoots, the aim of this work was to study the effects of Fe deficiency on the stem proteome of Medicago truncatula. Two-dimensional electrophoresis separation of stem protein extracts resolved 276 consistent spots in the whole experiment. Iron deficiency in absence or presence of CaCO3 caused significant changes in relative abundance in 10 and 31 spots, respectively, and 80% of them were identified by mass spectrometry. Overall results indicate that Fe deficiency by itself has a mild effect on the stem proteome, whereas Fe deficiency in the presence of CaCO3 has a stronger impact and causes changes in a larger number of proteins, including increases in stress and protein metabolism related proteins not observed in the absence of CaCO3. Both treatments resulted in increases in cell wall related proteins, which were more intense in the presence of CaCO3. The increases induced by Fe-deficiency in the lignin per protein ratio and changes in the lignin monomer composition, assessed by pyrolysis-gas chromatography-mass spectrometry and microscopy, respectively, further support the existence of cell wall alterations. BIOLOGICAL SIGNIFICANCE: In spite of being essential for the delivery of nutrients to the shoots, our knowledge of stem responses to nutrient deficiencies is very limited. The present work applies 2-DE techniques to unravel the response of this understudied tissue to Fe deficiency. Proteomics data, complemented with mineral, lignin and microscopy analyses, indicate that stems respond to Fe deficiency by increasing stress and defense related proteins, probably in response of mineral and osmotic unbalances, and eliciting significant changes in cell wall composition. The changes observed are likely to ultimately affect solute transport and distribution to the leaves.