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Iron (Fe) deficiency in calcareous soils is a significant agricultural challenge, affecting crop productivity and nutritional quality. This study aimed to isolate, characterize, and evaluate Fe solubilizing rhizobacterial isolates from maize rhizosphere in calcareous soils as potential biofertilizers. Forty bacterial isolates coded as SG1, SG2, …, SG40 were isolated and screened for siderophore production, with ten showing significant Fe solubilizing capabilities. These isolates were further assessed for phosphate solubilization and exopolysaccharides production. The selected bacterial isolates were also screened under axenic conditions for their ability to improve maize growth. The isolates SG8, SG13, SG24, SG30 and SG33 significantly enhanced growth parameters of maize. Notably, SG30 showed highest increment in shoot length (58%), root length (54%), root fresh and dry biomass (67% and 76%), SPAD value (67%), relative water contents (69%), root surface area (61%), and Fe concentration in shoots (79%) as compared to control. The biochemical characterization of these strains showed that all these strains have capability to solubilize insoluble phosphorus, produce indole-3-acetic acid (IAA), and ammonia with catalase, urease and protease activity. Molecular identification through 16s rRNA gene sequencing confirmed high similarity (99.7-100%) of the selected isolates to various Bacillus species, including B. pyramidoids, B. firmicutes, and B. cereus. The study provides a strong base for developing eco-friendly, cost-effective biofertilizers to address Fe deficiency in crops and promote sustainable agriculture., (© 2024. The Author(s).)

Iron (Fe) biofortification of edible organs without influencing crop yield is challenging, and potential solutions are largely unknown. Recently, Yan et al. identified a key regulator NAC78 (NAM/ATAF/CUC DOMAIN TRANSCRIPTION FACTOR 78) that enriches Fe in maize kernels without compromising crop yield. This may provide new crop yield management strategies for Fe acquisition and nutritional security., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Soil contamination with chromium (Cr) is becoming a primary ecological and health concern, specifically in the Kasur and Sialkot regions of Pakistan. The main objective of the current study was to evaluate the impact of foliar application of zinc oxide nanoparticles (ZnO NPs) (0, 25, 50, 100 mg L -1 ) and Fe NPs (0, 5, 10, 20 mg L -1 ) in red sails lettuce plants grown in Cr-contaminated soil. Our results showed that both ZnO and Fe NPs improved plant growth, and photosynthetic attributes by minimizing oxidative stress in lettuce plants through the stimulation of antioxidant enzyme activities. At ZnO NPs (100 mgL -1 ), dry weights of shoots and roots and fresh weights of shoots and roots were improved by 53%, 58%, 34%, and 45%, respectively, as compared to the respective control plants. The Fe NPs treatment (20 mgL -1 ) increased the dry weight of shoots and the roots and fresh weights of shoots and roots by 53%, 76%, 42%, and 70%, respectively. Application of both NPs reduced the oxidative stress caused by Cr, as evident by the findings of the current study, i.e., at the ZnO NPs (100 mgL -1 ) and Fe NPs (20 mgL -1 ), the EL declined by 32% and 44%, respectively, in comparison with respective control plants. Moreover, Fe and ZnO NPs enhanced the Fe and Zn contents in red sails lettuce plants. Application of ZnO NPs at 100 mg L -1 and Fe NPs at 20 mg L -1 , improved the Zn and Fe contents in plant leaves by 86%, and 68%, respectively, as compared to the control plants. This showed that the exogenous application of these NPs helped in Zn and Fe fortification in plants. At similar of concenteration ZnO NPs, CAT and APX activities were improved by 52% and 53%, respectively. Similarly, the POD contents were improved by 17% and 45% at 5 and 10 mg/L of Fe NPs. Furthermore, ZnO and Fe NPs limited the Cr uptake by plants, and the concentration of Cr in the leaves of lettuce was under the threshold limit. The exogenous application of ZnO NPs (100 mg L -1 ) and Fe NPs (20 mg L -1 ) reduced the Cr uptake in the leaves of red sails lettuce by 57% and 51%, respectively. In conclusion, ZnO and Fe NPs could be used for the improvement of plant growth and biomass as well as nutrient fortification in stressed environments. These findings not only underscore the efficacy of nanoparticle-assisted phytoremediation but also highlight its broader implications for sustainable agriculture and environmental health. However, future studies on other crops with molecular-level investigations are recommended for the validation of the results.

Micronutrient deficiency conditions, such as anemia, are the most prevalent global health problem due to inadequate iron and folate in dietary sources. Biofortification advancements can propel the rapid amelioration of nutritionally beneficial components in crops that are required to combat the adverse effects of micronutrient deficiencies on human health. To date, several strategies have been proposed to increase micronutrients in plants to improve food quality, but very few approaches have intrigued `clustered regularly interspaced short palindromic repeats' (CRISPR) modules for the enhancement of iron and folate concentration in the edible parts of plants. In this review, we discuss two important approaches to simultaneously enhance the bioavailability of iron and folate concentrations in rice endosperms by utilizing advanced CRISPR-Cas9-based technology. This includes the 'tuning of cis-elements' and 'enhancer re-shuffling' in the regulatory components of genes that play a vital role in iron and folate biosynthesis/transportation pathways. In particular, base-editing and enhancer re-installation in native promoters of selected genes can lead to enhanced accumulation of iron and folate levels in the rice endosperm. The re-distribution of micronutrients in specific plant organs can be made possible using the above-mentioned contemporary approaches. Overall, the present review discusses the possible approaches for synchronized iron and folate biofortification through modification in regulatory gene circuits employing CRISPR-Cas9 technology.

This commentary describes recent research discovering that the NAC transcription factor gene ZmNAC78 controls iron intake in maize and its implications for biofortification of this important crop. Using ZmNAC78, iron levels in maize can be more than doubled compared with current varieties., (© 2024 Institute of Botany, Chinese Academy of Sciences.)

Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper:

The vacuolar iron transporter (VIT) family is responsible for absorbing and storing iron ions in vacuoles. Here, the BnVIT-L2 gene from Brassica napus has been cloned for the first time and was found to be expressed in multiple tissues and organs, induced by iron stress. The BnVIT-L2 protein is located in vacuolar membranes and has the ability to bind both iron and other bivalent metal ions. Over-expression of the BnVIT-L2 gene increased lateral root number and main root length, as well as chlorophyll and iron content in transgenic Arabidopsis plants (BnVIT-L2/At) exposed to iron stress, compared to wild type Col-0. Furthermore, over-expression of this gene improved the adaptability of transgenic B. napus plants (BnVIT-L2-OE) under iron stress. The regulation of plant tolerance under iron stress by BnVIT-L2 gene may involve in the signal of reactive oxygen species (ROS), as suggested by Ribosome profiling sequencing (Ribo-seq). This study provides a reference for investigating plant growth and biofortification under iron stress through the BnVIT-L2 gene., Competing Interests: Declaration of competing interest The authors declare that they have no conflict of interest., (Copyright © 2024 Elsevier Masson SAS. All rights reserved.)

Efficient micronutrient acquisition is a critical factor in selecting micronutrient dense crops for human consumption. Enhanced exudation and re-uptake of metal chelators, so-called phytosiderophores, by roots of graminaceous plants has been implicated in efficient micronutrient acquisition. We compared PS biosynthesis and exudation as a response mechanism to either Fe, Zn or Cu starvation. Two barley (Hordeum vulgare L.) lines with contrasting micronutrient grain yields were grown hydroponically and PS exudation (LC-MS) and root gene expression (RNAseq) were determined after either Fe, Zn, or Cu starvation. The response strength of the PS pathway was micronutrient dependent and decreased in the order Fe > Zn > Cu deficiency. We observed a stronger expression of PS pathway genes and greater PS exudation in the barley line with large micronutrient grain yield suggesting that a highly expressed PS pathway might be an important trait involved in high micronutrient accumulation. In addition to several metal specific transporters, we also found that the expression of IRO2 and bHLH156 transcription factors was not only induced under Fe but also under Zn and Cu deficiency. Our study delivers important insights into the role of the PS pathway in the acquisition of different micronutrients., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have influenced the work reported in this paper., (Copyright © 2023 The Authors. Published by Elsevier B.V. All rights reserved.)

Iron (Fe) deficiency remains widespread among people in developing countries. To help solve this problem, breeders have been attempting to develop maize cultivars with high yields and high Fe concentrations in the kernels. We conducted a genome-wide association study and identified a gene, ZmNAC78 ( NAM/ATAF/CUC DOMAIN TRANSCRIPTION FACTOR 78 ), that regulates Fe concentrations in maize kernels. We cultivated maize varieties with both high yield and high Fe concentrations in their kernels by using a molecular marker developed from a 42-base pair insertion or deletion (indel) in the promoter of ZmNAC78 . ZmNAC78 expression is enriched in the basal endosperm transfer layer of kernels, and the ZmNAC78 protein directly regulates messenger RNA abundance of Fe transporters. Our results thus provide an approach to develop maize varieties with Fe-enriched kernels.

Cobalt (Co) is considered an essential element in agriculture as it is an important constituent of vitamin B 12 . Due to natural and anthropogenic factors, heavy metals, especially Co, accumulate in agricultural fields, but their high exposure produces ramifications in crop plants, thereby reducing crop yield and biomass. Excessive Co in plants causes oxidative stress, and as the stress progresses, Co competes with iron (Fe) thereby decreasing chlorophyll content and resulting in Fe deficiency in plants. A major concern is to counter the Co toxicity. Therefore, the current study aimed to mitigate Co-stress or Co-toxicity by using siderophore producing microbes and simultaneously mobilize Co and iron (Fe) in required amounts. In this study, 250 bacteria were isolated from agricultural and non-agricultural soils and screened for siderophore production. Initial siderophore screening revealed that 28.8% of the isolates produced siderophore. Subsequent screening for Co-tolerance showed that 16 isolates were tolerant to up to 20,000 ppm of Co and produced ACC deaminase, siderophore (96.82-99.67%), indole-3-acetic acid (15.15-70.55 µg/mL) and phosphate solubilisation (39.33-142.67 µg/mL). A plate assay (200 mM Co stress) revealed that four isolates (KSBTS 12, SBTS 12, CWTS 5 and CWTS 10) enhanced the growth of black gram (Vigna mungo L.). Furthermore, evaluation in pot studies (2000 ppm Co stress) revealed enhanced root (60.69-174.24%) and shoot length (3.27-143.96%) compared to the control. Inoculated plants also enhanced the uptake of nitrogen (37.33-42.36 mg/g) and phosphorous (3.12-3.92 mg/g), chlorophyll content (7.60-22.97 mg/g), siderophore quantity in the soils (282.41-331.53%) and the soil respiration activity such as hydrolysis of fluorescein diacetate (11.33-24.88 µg/g), dehydrogenase enzyme (109.76-197.26 µg/g) and alkaline phosphatase (631.53-918.20 µg/g). In conclusion, CWTS 5 (Bacillus subtilis) and CWTS 10 (Bacillus albus) can be used to mitigate Co-stress and mobilize Co and Fe in plants., (© 2023. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Post-harvest handling can affect micronutrient retention in biofortified crops through to the point of consumption. Here we conduct a systematic review identifying 67 articles examining the retention of micronutrients in conventionally bred biofortified maize, orange sweet potato, cassava, pearl millet, rice, beans and wheat. Provitamin A crops maintain high amounts compared with non-biofortified counterparts. Iron and zinc crops have more variability in micronutrient retention dependent on processing method; for maximum iron and zinc content, whole grain product consumption such as whole wheat flour or only slightly milled brown rice is beneficial. We offer preliminary suggestions for households, regulatory bodies and programme implementers to increase consumer awareness on best practices for preparing crops to maximize micronutrient content, while highlighting gaps in the literature. Our online, interactive Micronutrient Retention Dashboard ( https://www.cpnh.cornell.edu/mn-retention-db ) offers an at-a-glance view of the compiled minimum and maximum retention found, organized by processing method., (© 2023. The Author(s).)

In China, iron (Fe) availability is low in most soils but cadmium (Cd) generally exceeds regulatory soil pollution limits. Thus, biofortification of Fe along with mitigation of Cd in edible plant parts is important for human nutrition and health. Carbon dots (CDs) are considered as potential nanomaterials for agricultural applications. Here, Salvia miltiorrhiza-derived CDs are an efficient modulator of Fe, manganese (Mn), zinc (Zn), and Cd accumulation in plants. CDs irrigation (1 mg mL -1 , performed every week starting at the jointing stage for 12 weeks) increased Fe content by 18% but mitigated Cd accumulation by 20% in wheat grains. This finding was associated with the Fe 3+ -mobilizing properties of CDs from the soil and root cell wall, as well as endocytosis-dependent internalization in roots. The resulting excess Fe signaling mitigated Cd uptake via inhibiting TaNRAMP5 expression. Foliar spraying of CDs enhanced Fe (44%), Mn (30%), and Zn (19%) content with an unchanged Cd accumulation in wheat grains. This result is attributed to CDs-enhanced light signaling, which triggered shoot-to-root Fe deficiency response. This study not only reveals the molecular mechanism underlying CDs modulation of Fe signaling in plants but also provides useful strategies for concurrent Fe biofortification and Cd mitigation in plant-based foods., (© 2023 Wiley-VCH GmbH.)

Iron deficiency is still widespread as a major health problem even in countries with adequate food supply. It mainly affects women but also vegans, vegetarians, and athletes and can lead to various clinical pictures. Biofortification of vitamin C-rich vegetables with iron may be one new approach to face this nutritional challenge. However, so far, little is known about the consumer acceptance of iron-biofortified vegetables, particularly in developed countries. To address this issue, a quantitative survey of 1000 consumers in Germany was conducted. The results showed that depending on the type of vegetable, between 54% and 79% of the respondents were interested in iron-biofortified vegetables. Regression analysis showed a relationship between product acceptance, gender, and area of residence. In addition, relationships were found between consumer preferences for enjoyment, sustainability, and naturalness. Compared to functional food and dietary supplements, 77% of respondents would prefer fresh iron-rich vegetables to improve their iron intake. For a market launch, those iron-rich vegetables appear especially promising, which can additionally be advertised with claims for being rich in vitamin C and cultivated in an environmentally friendly way. Consumers were willing to pay EUR 0.10 to EUR 0.20 more for the iron-biofortified vegetables.

Nicotianamine (NA) is produced by NA synthase (NAS), which contains three genes in rice and is responsible for chelating metals such as iron (Fe) and zinc (Zn), as well as preserving metal homeostasis. In this study, we generated a transgenic plant ( 23D ) that shows simultaneous activation of OsNAS2 and OsNAS3 by crossing two previously identified activation-tagged mutants, OsNAS2-D1 ( 2D ) and OsNAS3-D1 ( 3D ). Concomitant activation of both genes resulted in the highest Fe and Zn concentrations in shoots and roots of the 23D plants grown under normal conditions and Fe and Zn limited growth conditions. Expression of genes for the biosynthesis of mugineic acid family phytosiderophores (MAs) and Fe and Zn uptake were enhanced in 23D roots. Additionally, 23D plants displayed superior growth to other plants at higher pH levels. Importantly, 23D seeds had NA and 2'-deoxymugineic acid (DMA) concentrations that were 50.6- and 10.0-fold higher than those of the WT. As a result, the mature grain Fe and Zn concentrations of the 23D plant were 4.0 and 3.5 times greater, respectively, than those of the WT. Furthermore, 23D plants exhibited the greatest resistance to excess metals. Our research suggests that simultaneous activation of OsNAS2 and OsNAS3 can enhance Fe and Zn accumulation in rice grains while also increasing plant tolerance to growing situations with metal deficiency and excess metal availability.

Plant-based foods provide all the crucial nutrients for human health. Among these, iron (Fe) is one of the essential micronutrients for plants and humans. A lack of Fe is a major limiting factor affecting crop quality, production, and human health. There are people who suffer from various health problems due to the low intake of Fe in their plant-based foods. Anemia has become a serious public health issue due to Fe deficiency. Enhancing Fe content in the edible part of food crops is a major thrust area for scientists worldwide. Recent progress in nutrient transporters has provided an opportunity to resolve Fe deficiency or nutritional problems in plants and humans. Understanding the structure, function, and regulation of Fe transporters is essential to address Fe deficiency in plants and to improve Fe content in staple food crops. In this review, we summarized the role of Fe transporter family members in the uptake, cellular and intercellular movement, and long-distance transport of Fe in plants. We draw insights into the role of vacuolar membrane transporters in the crop for Fe biofortification. We also provide structural and functional insights into cereal crops' vacuolar iron transporters (VITs). This review will help highlight the importance of VITs for improving the Fe biofortification of crops and alleviating Fe deficiency in humans.

Over three billion people suffer from various health issues due to the low supply of zinc (Zn) and iron (Fe) in their food. Low supply of micronutrients is the main cause of malnutrition and biofortification could help to solve this issue. Understanding the molecular mechanisms of biofortification is challenging. The membrane transporters are involved in the uptake, transport, storage, and redistribution of Zn and Fe in plants. These transporters are also involved in biofortification and help to load the Zn and Fe into the endosperm of the seeds. Very little knowledge is available on the role and functions of membrane transporters involved in seed biofortification. Understanding the mechanism and role of membrane transporters could be helpful to improve biofortification. In this review, we provide the details on membrane transporters involved in the uptake, transport, storage, and redistribution of Zn and Fe. We also discuss available information on transporters involved in seed biofortification. This review will help plant breeders and molecular biologists understand the importance and implications of membrane transporters for seed biofortification., (© 2022. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Microgreens are foods with high nutritional value, which can be further enhanced with biofortification. Crop biofortification involves increasing the accumulation of target nutrients in edible plant tissues through fertilization or other factors. The purpose of the present study was to evaluate the potential for biofortification of some vegetable microgreens through iron (Fe) enrichment. The effect of nutrient solution supplemented with iron chelate (1.5, 3.0 mg/L) on the plant's growth and mineral concentration of purple kohlrabi, radish, pea, and spinach microgreens was studied. Increasing the concentration of Fe in the medium increased the Fe content in the leaves of the species under study, except for radish. Significant interactions were observed between Fe and other microelements (Mn, Zn, and Cu) content in the shoots. With the increase in the intensity of supplementation with Fe, regardless of the species, the uptake of zinc and copper decreased. However, the species examined suggested that the response to Fe enrichment was species-specific. The application of Fe didn't influence plant height or fresh and dry weight. The chlorophyll content index (CCI) was different among species. With increasing fertilisation intensity, a reduction in CCI only in peas resulted. A higher dose of iron in the medium increased the fluorescence yield of spinach and pea microgreens. In conclusion, the tested species, especially spinach and pea, grown in soilless systems are good targets to produce high-quality Fe biofortified microgreens.

Background: In neutral or alkaline soils, iron (Fe) easily forms insoluble complexes, which makes it difficult for plants to utilize Fe in the soil for nutrition. Polyester sulfur-coated urea (PSCU) is a novel controlled-release fertilizer widely used in China and some foreign countries, and it has been proven that sulfur film from controlled-release fertilizers can significantly improve the activation of Fe and other elements in the soil. However, few studies have focused on the effects of PSCU application on Fe accumulation in rice grain in alkaline soils., Results: Both our field and pot experiments proved that PSCU application could significantly improve rice grain yield and Fe concentration in brown rice in alkaline soil. This effect differs with different types of alkaline soils (i.e. medium-saline, sandy soil and/or silt soil). PSCU is released slowly, and the release rate is different in different alkaline soils. Rice shoot nitrogen (N) uptake was significantly enhanced with PSCU application., Conclusion: The results suggested that PSCU application in alkaline soils could significantly enhance brown rice Fe concentration and production. This effect differed with different kinds of alkaline soils. The study identified some efficient fertilizers to improve the Fe status in alkaline soils. © 2021 Society of Chemical Industry., (© 2021 Society of Chemical Industry.)

Background: Zinc-biofortified rice could contribute to zinc intake in deficient populations, but processing it into parboiled rice could affect this potential benefit. Zinc and iron true retention (TR) in milled rice produced under conditions resembling household and commercial parboiled methods was evaluated. Zinc and iron TR in milled rice obtained from biofortified and non-biofortified rice subjected to different soaking temperatures during parboiling was also evaluated., Results: Conditions resembling commercial parboiling methods resulted in 52.2-59.7% zinc TR and 55.4-79.1% iron TR, whereas those used for household parboiling resulted in 70.7-79.6% zinc TR and 78.2-119.8% iron TR. Zinc TR in milled (8-16% bran removal) biofortified and non-biofortified parboiled rice was 50.6-66.8% when soaking rough rice at 20 °C and 29.9-56.0% when soaking rough rice at 65 °C; both had lower zinc TR than non-parboiled rice (58.0-80.6%). Iron TR was generally similar between milled non-parboiled and parboiled rice (26.2-67.6%) and between parboiled biofortified and non-biofortified milled rice., Conclusion: Parboiling conditions used to obtain milled rice targeted for own household consumption resulted in higher zinc and iron TR compared to parboiling conditions used for milled rice targeted for markets. More zinc from the inner endosperm moved towards the outer layers at high soaking temperature, resulting in lower zinc TR for milled parboiled rice soaked in hotter water. Parboiled rice soaked at temperatures used in households could provide more zinc to diets compared to rice soaked in hotter water commonly used in large rice mills, especially when rice is extensively milled. © 2021 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry., (© 2021 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.)

This work presents new findings on the nutritional quality of recently introduced biofortified and non-biofortified cowpea cultivars as well as some common beans. ICP-MS was used for the measurements. Biofortified cowpea cultivars showed high levels of Fe and Zn, greater than 60 and 40 mg kg -1 dry weight, respectively. The in vitro digestion protocol enabled simultaneous evaluation of bioaccessibility and bioavailability. Fe levels in cowpea cultivars were ca. 2.5-fold higher than in common beans. Cowpea seeds also had higher Zn levels, reaching 50.1% bioaccessibility and 44.2% bioavailability. Cooking improved the availability of micronutrients in bean seeds. The cooked biofortified Aracê cowpea showed a high Zn bioavailability above 60%. Consumption of 50 g of Aracê would contribute 27% and 48% of the Fe and Zn DRI for 1-3-year-old children. The new cowpea cultivars biofortified are a potential vehicle for improving the Fe and Zn status in groups in which the micronutrient deficiency is prevalent., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Iron deficiency anemia affects approximately one-third of the world's population, and about half the cases are due to iron deficiency. The latest research on iron metabolism published in original articles and systematic reviews is described, and references to recent reviews provided. The topics include dietary sources and bioavailability, iron homeostasis, functions of iron in the body, and biomarkers of status. The consequences of iron deficiency and excess are discussed, with particular focus on vulnerable populations such as pregnant women, infants and the elderly. The newest dietary recommendations, including dietary reference values and food based dietary guidelines, are briefly summarized, followed by the latest developments in food fortification and iron supplementation., (Copyright © 2021 Elsevier Inc. All rights reserved.)

Background: Sweetpotato and potato are fast-maturing staple crops and widely consumed in low- and middle-income countries. Conventional breeding to biofortify these crops with iron could improve iron intakes. To our knowledge, iron absorption from sweetpotato and potato has not been assessed., Objective: The aim was to assess iron absorption from regular and iron-biofortified orange-fleshed sweetpotato in Malawi and yellow-fleshed potato and iron-biofortified purple-fleshed potato in Peru., Methods: We conducted 2 randomized, multiple-meal studies in generally healthy, iron-depleted women of reproductive age. Malawian women (n = 24) received 400 g regular or biofortified sweetpotato test meals and Peruvian women (n = 35) received 500 g regular or biofortified potato test meals. Women consumed the meals at breakfast for 2 wk and were then crossed over to the other variety. We labeled the test meals with 57Fe or 58Fe and measured cumulative erythrocyte incorporation of the labels 14 d after completion of each test-meal sequence to calculate iron absorption. Iron absorption was compared by paired-sample t tests., Results: The regular and biofortified orange-fleshed sweetpotato test meals contained 0.55 and 0.97 mg Fe/100 g. Geometric mean (95% CI) fractional iron absorption (FIA) was 5.82% (3.79%, 8.95%) and 6.02% (4.51%, 8.05%), respectively (P = 0.81), resulting in 1.9-fold higher total iron absorption (TIA) from biofortified sweetpotato (P < 0.001). The regular and biofortified potato test meals contained 0.33 and 0.69 mg Fe/100 g. FIA was 28.4% (23.5%, 34.2%) from the regular yellow-fleshed and 13.3% (10.6%, 16.6%) from the biofortified purple-fleshed potato meals, respectively (P < 0.001), resulting in no significant difference in TIA (P = 0.88)., Conclusions: FIA from regular yellow-fleshed potato was remarkably high, at 28%. Iron absorbed from both potato test meals covered 33% of the daily absorbed iron requirement for women of reproductive age, while the biofortified orange-fleshed sweetpotato test meal covered 18% of this requirement. High polyphenol concentrations were likely the major inhibitors of iron absorption. These trials were registered at www.clinicaltrials.gov as NCT03840031 (Malawi) and NCT04216030 (Peru)., (© The Author(s) 2020. Published by Oxford University Press on behalf of the American Society for Nutrition.)

The micronutrients iron (Fe) and zinc (Zn) are not only essential for plant survival and proliferation but are crucial for human health. Increasing Fe and Zn levels in edible parts of plants, known as biofortification, is seen a sustainable approach to alleviate micronutrient deficiency in humans. Wheat, as one of the leading staple foods worldwide, is recognized as a prioritized choice for Fe and Zn biofortification. However, to date, limited molecular and physiological mechanisms have been elucidated for Fe and Zn homeostasis in wheat. The expanding molecular understanding of Fe and Zn homeostasis in model plants is providing invaluable resources to biofortify wheat. Recent advancements in NGS (next generation sequencing) technologies coupled with improved wheat genome assembly and high-throughput genotyping platforms have initiated a revolution in resources and approaches for wheat genetic investigations and breeding. Here, we summarize molecular processes and genes involved in Fe and Zn homeostasis in the model plants Arabidopsis and rice, identify their orthologs in the wheat genome, and relate them to known wheat Fe/Zn QTL (quantitative trait locus/loci) based on physical positions. The current study provides the first inventory of the genes regulating grain Fe and Zn homeostasis in wheat, which will benefit gene discovery and breeding, and thereby accelerate the release of Fe- and Zn-enriched wheats.

The future of food and sustainability of the staple food crops are of utmost importance in the 21st century. Micronutrient deficiency, for example, in iron and zinc, is a common cause of human diseases. Mineral content of the staple food crops has therefore crosscutting importance for food engineering and planetary health. Finger millet, a staple food of agricultural importance worldwide, is rich in iron and zinc, and an ideal model to study the prospects of biofortified foods in times of climate change. We report here a multiomics study of the iron and zinc homeostasis in the finger millet. We identified and characterized 15 candidate genes potentially involved in iron and zinc homeostasis pathways in the finger millet. Structural and functional annotation of the candidate genes revealed a high similarity index with their respective homologs ( Oryza sativa , Triticum aestivum , Zea mays , Hordeum vulgare , and Setaria italica ). Transcriptome-wide expression analysis showed that genes involved in uptake and translocation of iron and zinc are highly expressed in the GP-1 genotype, while those involved in bioavailability of iron and zinc are expressed more in the GP-45 genotype of the finger millet. In conclusion, finger millet, being a stress-resilient crop, utilizes a combination of strategies in iron and zinc homeostasis pathway, which appear to play an important role in food crop acquisition of iron and zinc, despite environmentally limiting conditions. These data offer molecular insights on iron and zinc accumulation and paves the way for new strategies toward staple food crop with mineral biofortification.

Background: The predominant bean iron (Fe) biofortification approach is to breed for high Fe concentration and assumes the average Fe concentration is 50 μg/g. This approach also assumes that a 40 μg/g increase is sustainable and Fe bioavailability will not decrease to negate the increase in Fe., Objective: The overall objective was to determine if bean Fe biofortification via breeding for high Fe is producing beans with higher Fe concentration relative to nonbiofortified lines found in the East Africa marketplace., Methods: Seventy-six marketplace samples (East Africa Marketplace Collection; EAMC), and 154 genotypes known to be representative of the marketplace were collected from breeders in the Pan-Africa Bean Research Alliance (designated the East Africa Breeder Collection; EABC). Within the EAMC and EABC were 18 and 35 samples, respectively, that were released as biofortified lines. All samples were measured for Fe concentration. The Caco-2 cell bioassay assessed Fe bioavailability of the EAMC. Biofortified versus nonbiofortified samples were compared by the appropriate t-test or ANOVA., Results: The Fe concentration of the 58 nonbiofortified EAMC lines was (mean ± SD [range]) 71 ± 9 μg/g (52-93 μg/g) which did not differ significantly from the 18 biofortified EAMC varieties (71 ± 11 μg/g [55-94 μg/g]). The Fe concentration of the 119 nonbiofortified EABC varieties was 66 ± 7 μg/g (51-90 μg/g) which was significantly different (P < 0.0001) from the 35 EABC biofortified lines (73 ± 9 μg/g [60-91 μg/g]). However, the EABC biofortified lines were not different from the nonbiofortified EAMC samples. In the Caco-2 cell bioassay, biofortified EAMC varieties did not deliver more Fe compared with nonbiofortified EAMC varieties., Conclusions: The assumptions of the high Fe bean biofortification approach are not met in the East African marketplace. Iron concentration and bioavailability measurement indicate the biofortified bean varieties are providing no additional dietary Fe., (Published by Oxford University Press on behalf of the American Society for Nutrition 2020.)

The deep relationship between plants and humans predates civilization, and our reliance on plants as sources of food, feed, fiber, fuels, and pharmaceuticals continues to increase. Understanding how plants grow and overcome challenges to their survival is critical for using these organisms to meet current and future demands for food and other plant-derived materials. This thematic review series on "plants in the real world" presents a set of eight reviews that highlight advances in understanding plant health, including the role of thiamine (vitamin B 1 ), iron, and the plant immune system; how plants use ethylene and ubiquitin systems to control growth and development; and how new gene-editing approaches, the redesign of plant cell walls, and deciphering herbicide resistance evolution can lead to the next generation of crops., Competing Interests: Conflict of interest—The author declares that he has no conflicts of interest with the contents of this article., (© 2020 Jez.)

Breeding and engineering of biofortified crops will benefit from a better understanding of bottlenecks controlling micronutrient loading within the seeds. However, few studies have addressed the changes in micronutrient concentrations, localization, and speciation occurring over time. Therefore, we studied spatial patterns of zinc and iron accumulation during grain development in two barley lines with contrasting grain zinc concentrations. Microparticle-induced-X-ray emission and laser ablation-inductively coupled plasma mass spectrometry were used to determine tissue-specific accumulation of zinc, iron, phosphorus, and sulfur. Differences in zinc accumulation between the lines were most evident in the endosperm and aleurone. A gradual decrease in zinc concentrations from the aleurone to the underlying endosperm was observed, while iron and phosphorus concentrations decreased sharply. Iron co-localized with phosphorus in the aleurone, whereas zinc co-localized with sulfur in the sub-aleurone. We hypothesize that differences in grain zinc are largely explained by the endosperm storage capacity. Engineering attempts should be targeted accordingly.

Background: Iron deficiency is a major public health concern in Ethiopia, where the traditional diet is based on tef injera. Iron absorption from injera is low due to its high phytic acid (PA) content., Objectives: We investigated ways to increase iron absorption from FeSO4-fortified tef injera in normal-weight healthy women (aged 21-29 y)., Methods: Study A (n = 22) investigated the influence on fractional iron absorption (FIA) from FeSO4-fortified injera of 1) replacing 10% tef flour with whole wheat flour (a source of wheat phytase), or 2) adding an isolated phytase from Aspergillus niger. Study B (n = 18) investigated the influence on FIA of replacing FeSO4 in tef injera with different amounts of NaFeEDTA. In both studies, the iron fortificants were labeled with stable isotopes and FIA was calculated from erythrocyte incorporation of stable iron isotopes 14 d after administration., Results: In study A, the median (IQR) FIA from the 100% tef injera meal was 1.5% (0.7-2.8%). This increased significantly (P < 0.05) to 5.3% (2.4-7.1%) on addition of 10% whole wheat flour, and to 3.6% (1.6-6.2%) on addition of A. niger phytase. PA content of the 3 meals was 0.62, 0.20, and 0.02 g/meal, respectively. In study B, the median (IQR) FIA from the 100% tef injera meal was 3.3% (1.1-4.4%) and did not change significantly (P > 0.05) on replacing 50% or 75% of FeSO4 with NaFeEDTA., Conclusions: FIA from tef injera by young women was very low. NaFeEDTA was ineffective at increasing iron absorption, presumably due to the relatively low EDTA:Fe molar ratios. Phytate degradation, however, greatly increased during tef fermentation on addition of native or isolated phytases. Replacing 10% tef with whole wheat flour during injera fermentation tripled FIA in young women and should be considered as a potential strategy to improve iron status in Ethiopia., (Copyright © The Author(s) on behalf of the American Society for Nutrition 2020.)

Rice is critical for sustainable food and nutritional security; however, nominal micronutrient quantities in grains aggravate malnutrition in rice-eating poor populations. In this study, we evaluated genetic diversity in grain iron (Fe) and zinc (Zn) contents using trait-linked simple sequence repeat (SSR) markers in the representative subset of a large collection of local and exotic rice germplasm. Results demonstrated that aromatic fine grain accessions contained relatively higher Fe and Zn contents in brown rice (BR) than coarse grain accessions and a strong positive correlation between both mineral elements. Genotyping with 24 trait-linked SSR markers identified 21 polymorphic markers, among which 17 demonstrated higher gene diversity and polymorphism information content (PIC) values, strongly indicating that markers used in current research were moderate to highly informative for evaluating the genetic diversity. Population structure, principal coordinate and phylogenetic analyses classified studied rice accessions into two fine grain specific and one fine and coarse grain admixture subpopulations. Single marker analysis recognized four ZnBR and single FeBR significant marker-trait associations (MTAs) contributing 15.41-39.72% in total observed phenotypic variance. Furthermore, high grain Fe and Zn contents linked marker alleles from significant MTAs were also identified. Collectively, these results indicate a wide genetic diversity exist in grain Fe and Zn contents of studied rice accessions and reveal perspective for marker-assisted biofortification breeding., Competing Interests: The authors have declared that no competing interests exist.

Background: Nutrient deficiency in humans, especially in children and lactating women, is a major concern. Increasing the micronutrient concentration in staple crops like rice is one way to overcome this. The micronutrient content in rice, especially the iron (Fe) and zinc (Zn) content, is highly variable. The identification of rice genotypes in which there are naturally high Fe and Zn concentrations across environments is an important target towards the production of biofortified rice., Results: Phenotypic correlations between grain Fe and Zn content were positive and significant in all environments but a significant negative association was observed between grain yield and grain Fe and Zn. Promising breeding lines with higher Zn or Fe content, or both, were: IR 82475-110-2-2-1-2 (Zn: 20.24-37.33 mg kg -1 ; Fe: 7.47-14.65 mg kg -1 ); IR 83294-66-2-2-3-2 (Zn: 22-37-41.97 mg kg -1 ; Fe: 9.43-17.16); IR 83668-35-2-2-2 (Zn: 27.15-42.73 mg kg -1 ; Fe: 6.01-14.71); IR 68144-2B-2-2-3-1-166 (Zn: 23.53-40.30 mg kg -1 ; Fe: 10.53-17.80 mg kg -1 ) and RP Bio 5478-185M7 (Zn: 22.60-40.07 mg kg -1 ; Fe: 7.64-14.73 mg kg -1 ). Among these, IR82475-110-2-2-1-2 (Zn: 20.24-37.33 mg kg -1 ; Fe: 7.47-14.65 mg kg -1 ) is also high yielding with 3.75 t ha -1 . Kelhrie Cha (Zn: 17.76-36.45 mg kg -1 ; Fe: 7.17-14.77 mg kg -1 ), Dzuluorhe (Zn: 17.48-39.68 mg kg -1 ; Fe: 7.89-19.90 mg kg -1 ), Nedu (Zn: 18.97-43.55 mg kg -1 Fe: 8.01-19.51 mg kg -1 ), Kuhusoi-Ri-Sareku (Zn: 17.37-44.14 mg kg -1 ; Fe: 8.99-14.30 mg kg -1 ) and Mima (Zn: 17.10-45.64 mg kg -1 ; Fe: 9.97-17.40 mg kg -1 ) were traditional donor genotypes that possessed both high grain Fe and high Zn content., Conclusion: Significant genotype × location (G × L) effects were observed in all traits except Fe. Genetic variance was significant and was considerably larger than the variance of G × L for grain Zn and Fe content traits, except grain yield. The G × L × year variance component was significant in all cases. © 2020 Society of Chemical Industry., (© 2020 Society of Chemical Industry.)

Basidiomycetes can bioaccumulate high iron contents, but there are few studies on iron availability from the mycelial biomass in order to support their use as an iron-enriched fungal food. This study aimed to evaluate the in vitro iron bioaccumulation and availability in the mycelial biomass of edible and medicinal basidiomycetes grown in two distinct culture media. Lentinus crinitus, Ganoderma lucidum, Schizophyllum commune, Pleurotus ostreatus, Pleurotus eryngii, Lentinula edodes, and Agaricus subrufescens were grown in liquid culture medium of malt extract or sugarcane molasses to obtain iron-bioaccumulated mycelial biomass. P. ostreatus was the fungus that most bioaccumulated iron, followed by S. commune, and P. eryngii; they also had the highest mycelial biomass growth and iron transfer from the culture medium to the mycelial biomass. Mycelial iron availability is species-specific, regardless of the culture medium and the iron bioaccumulation capacity of the fungus in the mycelial biomass. Mycelial biomass of S. commune, followed by G. lucidum, P. ostreatus, and P. eryngii, associated with molasses culture medium, are the best choice for the production of iron-enriched mycelial biomass.

Wheat (T. aestivum L.) is the second most important staple food crop consumed in the form of various end-use products across the world. However, it contains lower concentrations of Fe and Zn leading to micronutrient deficiency in human beings where wheat is the sole diet. Therefore, increasing grain Fe/Zn content in wheat has become priority in wheat breeding programmes across the world. Understanding the molecular mechanism of Fe/Zn transport and accumulation in grains is required to expedite the breeding process. For this purpose, whole seedling transcriptome analysis was conducted in four wheat genotypes (CRP 1660, Sonora 64, Vinata, : high, and DBW17: low) differing in grain Fe/Zn content under controlled and Fe/Zn deficient conditions. Twenty eight key transcripts involved in phytosiderophore biosynthesis, Fe/Zn uptake and transport were identified. Expression analysis of 12 of the transcripts using qPCR was conducted in seedling stage and flag leaf which exhibited greater differential accumulation in CRP 1660 followed by Vinata, Sonora 64 and DBW 17 in both flag leaf and seedling. However, there was significantly higher differential accumulation of the transcripts in flag leaf as compared to seedling. In CRP 1660, transcripts pertaining to phytosiderophore biosynthesis like DMAS1-B, NRAMP2 and NAAT2-D showed greater accumulation. Additionally, corresponding miRNAs were also identified for these 28 transcripts. The findings will help in better understanding of molecular basis of Fe/Zn transport and accumulation in grain and subsequent utilization in breeding to improve Fe/Zn content in wheat grain., Competing Interests: Declaration of Competing Interest The authors report no declarations of interest., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Background: Iron-biofortified staple foods can improve iron status and resolve iron deficiency. However, whether improved iron status from iron biofortification can improve physical performance remains unclear., Objective: This study aimed to examine whether changes in iron status from an iron-biofortified bean intervention affect work efficiency., Methods: A total of 125 iron-depleted (ferritin <20 μg/L) female Rwandan university students (18-26 y) were selected from a larger sample randomly assigned to consume iron-biofortified beans (Fe-Bean; 86.1 mg Fe/kg) or conventional beans (control: 50.6 mg Fe/kg) twice daily for 18 wk (average of 314 g beans consumed/d). Blood biomarkers of iron status (primary outcome) and physical work efficiency (secondary outcome) were measured before and after the intervention. Work performed was assessed during 5-min steady-state periods at 0-, 25-, and 40-W workloads using a mechanically braked cycle ergometer. Work efficiency was calculated at 25 W and 40 W as the work accomplished divided by the energy expended at that workload above that expended at 0 W. General linear models were used to evaluate the relation between changes in iron status biomarkers and work efficiency., Results: The Fe-Bean intervention had significant positive effects on hemoglobin, serum ferritin, and body iron stores but did not affect work efficiency. However, 18-wk change in hemoglobin was positively related to work efficiency at 40 W in the full sample (n = 119; estimate: 0.24 g/L; 95% CI: 0.01, 0.48 g/L; P = 0.044) and among women who were anemic (hemoglobin <120 g/L) at baseline (n = 43; estimate: 0.64 g/L; 95% CI: 0.05, 1.23 g/L; P = 0.036). Among women who were nonanemic at baseline, change in serum ferritin was positively related to change in work efficiency at 40 W (n = 60; estimate: 0.50 μg/L; 95% CI: 0.06, 0.95 μg/L; P = 0.027)., Conclusions: Increasing iron status during an iron-biofortified bean feeding trial improves work efficiency in iron-depleted, sedentary women. This trial was registered at clinicaltrials.gov as NCT01594359., (Copyright © The Author(s) 2020.)

Background: Iron is one of the nutrients that is essential for the human body. Despite the abundance of iron on earth, about two billion people worldwide are affected by iron deficiency. Iron biofortification of wheat, instead of supplementation and food fortification, provides a pragmatic approach to solve the problem of iron deficiency. In this study, 144 diverse wheat genotypes were evaluated for grain iron and yield potential, to estimate the potential for the iron biofortification of high-yielding wheat varieties., Results: Genotypes did not differ significantly across the species, but within species the differences were significant for grain iron content and the phytate:iron molar ratio. Triticum aestivum (bread wheat) had the highest yield potential with more diversity than other Triticum species. Genotypes with high iron contents were crossed with high-yielding genotypes in line × tester fashion to check the gene action controlling these traits. The combining ability analysis showed non-additive gene action controlling grain iron, grain phytate, and grain yield. Heterosis manifestation also indicated some transgressive segregates with high specific combining ability effects., Conclusion: There was considerable genetic potential for improving the grain iron content in the germplasm to provide an economical and long-lasting solution to benefit an iron-deficient population. Triticum aestivum had the highest variation and potential for iron biofortification. This study indicated the possibility of simultaneous improvement in grain iron and grain yield by producing a new variety through continuous selective breeding. © 2019 Society of Chemical Industry., (© 2019 Society of Chemical Industry.)

The objective of this study was to evaluate the interaction of pro-vitamin A-rich sweet potato on iron bioavailability of biofortified cowpeas, using in vitro Caco-2 cells and in vivo depletion-repletion rat model. Mixtures of conventional rice with cultivars of iron-biofortified (Aracê, Xiquexique, and Tumucumaque) or conventional (Guariba) cowpeas with or without sweet potato biofortified with pro-vitamin A carotenoids were evaluated. The ratio of ferritin/total protein in Caco-2 cells was used as the index of cellular Fe uptake in the in vitro assay. The animal study evaluated the hemoglobin gain, the relative biological value, and the gene expression of transferrin and ferritin proteins by reverse transcription polymerase chain reaction. In the in vitro study, Xiquexique cowpea presented higher bioavailability of iron in the absence of sweet potato, and no difference was observed between the other cultivars of cowpea with and without sweet potato. The in vivo bioavailability (relative biological value of hemoglobin regeneration efficiency) differed statistically only between Guariba groups added to sweet potato and Tumucumaque. Ferritin mRNA expression did not differ between the test and control (ferrous sulfate) groups. Regarding the transferrin mRNA expression, there was a difference between the test and control groups except for the Xiquexique group. The association of rice and beans with sweet potato rich in carotenoids favored the gene expression of proteins involved in the iron metabolism, as well as its bioavailability, corroborating beneficial effects of this mixture. Xiquexique cowpea was shown to be the most promising compared to the other cultivars, exhibiting higher iron content in the digestible fraction, better in vitro bioavailability of iron, and transferrin gene expression. PRACTICAL APPLICATION: Data from the study indicated greater in vitro bioavailability of iron for Xiquexique cowpea and sweet potato mixtures, in addition to the greater regeneration efficiency of hemoglobin in vivo as the bioavailability of iron among biofortified beans, highlighting the promising benefits of biofortification., (© 2020 Institute of Food Technologists®.)

Biofortification is an effective method to improve the nutritional content of crops and nutritional intake. Breeding for higher micronutrient mineral content in beans is correlated with an increase in phytic acid, a main inhibitor of mineral absorption in humans. Low phytic acid ( lpa ) beans have a 90% lower phytic acid content compared to conventional beans. This is the first study to investigate mineral and total phytic acid retention after preparing common household recipes from conventional, biofortified and lpa beans. Mineral retention was determined for two conventional, three biofortified and two lpa bean genotypes. Treatments included soaking, boiling (boiled beans) and refrying (bean paste). The average true retention of iron after boiling was 77.2-91.3%; for zinc 41.2-84.0%; and for phytic acid 49.9-85.9%. Soaking led to a significant decrease in zinc and total phytic acid after boiling and refrying, whereas for iron no significant differences were found. lpa beans did not exhibit a consistent pattern of difference in iron and phytic acid retention compared to the other groups of beans. However, lpa beans had a significantly lower retention of zinc compared to conventional and biofortified varieties ( p < 0.05). More research is needed to understand the underlying factors responsible for the differences in retention between the groups of beans, especially the low retention of zinc. Combining the lpa and biofortification traits could further improve the nutritional benefits of biofortified beans, by decreasing the phytic acid:iron and zinc ratio in beans., Competing Interests: The authors declare no conflict of interest.

Background: High yielding rice varieties are usually low in grain iron (Fe) and zinc (Zn) content. These two micronutrients are involved in many enzymatic activities, lack of which cause many disorders in human body. Bio-fortification is a cheaper and easier way to improve the content of these nutrients in rice grain., Results: A population panel was prepared representing all the phenotypic classes for grain Fe-Zn content from 485 germplasm lines. The panel was studied for genetic diversity, population structure and association mapping of grain Fe-Zn content in the milled rice. The population showed linkage disequilibrium showing deviation of Hardy-Weinberg's expectation for Fe-Zn content in rice. Population structure at K = 3 categorized the panel population into distinct sub-populations corroborating with their grain Fe-Zn content. STRUCTURE analysis revealed a common primary ancestor for each sub-population. Novel quantitative trait loci (QTLs) namely qFe3.3 and qFe7.3 for grain Fe and qZn2.2, qZn8.3 and qZn12.3 for Zn content were detected using association mapping. Four QTLs, namely qFe3.3, qFe7.3, qFe8.1 and qFe12.2 for grain Fe content were detected to be co-localized with qZn3.1, qZn7, qZn8.3 and qZn12.3 QTLs controlling grain Zn content, respectively. Additionally, some Fe-Zn controlling QTLs were co-localized with the yield component QTLs, qTBGW, OsSPL14 and qPN. The QTLs qFe1.1, qFe3.1, qFe5.1, qFe7.1, qFe8.1, qZn6, qZn7 and gRMm9-1 for grain Fe-Zn content reported in earlier studies were validated in this study., Conclusion: Novel QTLs, qFe3.3 and qFe7.3 for grain Fe and qZn2.2, qZn8.3 and qZn12.3 for Zn content were detected for these two traits. Four Fe-Zn controlling QTLs and few yield component QTLs were detected to be co-localized. The QTLs, qFe1.1, qFe3.1, qFe5.1, qFe7.1, qFe8.1, qFe3.3, qFe7.3, qZn6, qZn7, qZn2.2, qZn8.3 and qZn12.3 will be useful for biofortification of the micronutrients. Simultaneous enhancement of Fe-Zn content may be possible with yield component traits in rice.

To ensure the success of the new generation in annual species, the mother plant transfers a large proportion of the nutrients it has accumulated during its vegetative life to the next generation through its seeds. Iron (Fe) is required in large amounts to provide the energy and redox power to sustain seedling growth. However, free Fe is highly toxic as it leads to the generation of reactive oxygen species. Fe must, therefore, be tightly bound to chelating molecules to allow seed survival for long periods of time without oxidative damage. Nevertheless, when conditions are favorable, the seed's Fe stores have to be readily remobilized to achieve the transition toward active photosynthesis before the seedling becomes able to take up Fe from the environment. This is likely critical for the vigor of the young plant. Seeds constitute an important dietary source of Fe, which is essential for human health. Understanding the mechanisms of Fe storage in seeds is a key to improve their Fe content and availability in order to fight Fe deficiency. Seed longevity, germination efficiency and seedling vigor are also important traits that may be affected by the chemical form under which Fe is stored. In this review, we summarize the current knowledge on seed Fe loading during development, long-term storage and remobilization upon germination. We highlight how this knowledge may help seed Fe biofortification and discuss how Fe storage may affect the seed quality and germination efficiency., (© 2020 The Author(s). Published by Portland Press Limited on behalf of the Biochemical Society.)

Background: Cowpea (Vigna unguiculata L. Walph) is predominantly consumed in the North and Northeast regions of Brazil, and its biofortification with iron seeks to reduce the high prevalence of iron deficiency anemia in these regions. It is commonly eaten cooked; however, in the germinated form, it can improve nutritional quality by reducing the antinutritional factors and consequently improving the bioavailability of elements. The present study aimed to determine the physico-chemical characteristics, bioaccessibility and bioavailability of iron in biofortified germinated cowpea., Results: There was no statistical difference between the germinated and cooked beans with regard to centesimal composition. Germinated beans had phytates and tannins similar to cooked beans. The phytate-iron molar ratio for all groups did not present a statistical difference (cooking 3.58 and 3.41; germinated 3.94 and 3.51), nor did the parameters evaluating in vivo iron bioavailability. Total phenolics was higher in the germinated group (cooking 0.56 and 0.64; Germinated 2.05 and 2.45 mg gallic acid kg -1 ). In vitro bioaccessibility of iron of germinated beans presented higher values (P ≤ 0.05) compared to cooked beans. There was higher expression of divalent metal transporter-1 in biofortified and germinated beans., Conclusion: The iron bioavailability from the biofortified and germinated beans was comparable to ferrous sulfate. Germination can be considered as an alternative and efficient method for consuming cowpea, presenting good iron bioaccessibility and bioavailability. © 2019 Society of Chemical Industry., (© 2019 Society of Chemical Industry.)

Cu, Fe, Mn, Mo, Selenium (Se), and Zn bioavailability from selenate- and selenite-enriched lettuce plants was studied by in vitro gastrointestinal digestion followed by an assay with Caco-2 cells. The plants were cultivated in the absence and presence of two concentrations (25 and 40 µmol/L of Se). After 28 days of cultivation, the plants were harvested, dried, and evaluated regarding the total concentration, bioaccessibility, and bioavailability of the analytes. The results showed that biofortification with selenate leads to higher Se absorption by the plant than biofortification with selenite. For the other nutrients, Mo showed high accumulation in the plants of selenate assays, and the presence of any Se species led to a reduction of the plant uptake of Cu and Fe. The accumulation of Zn and Mn was not strongly influenced by the presence of any Se species. The bioaccessibility values were approximately 71%, 10%, 52%, 84%, 71%, and 86% for Cu, Fe, Mn, Mo, Se, and Zn, respectively, and the contribution of the biofortified lettuce to the ingestion of these minerals is very small (except for Se and Mo). Due to the low concentrations of elements from digested plants, it was not possible to estimate the bioavailability for some elements, and for Mo and Zn, the values are below 6.9% and 3.4% of the total concentration, respectively. For Se, the bioavailability was greater for selenite-enriched than selenate-enriched plants (22% and 6.0%, respectively), because selenite is biotransformed by the plant to organic forms that are better assimilated by the cells., (© 2019 Institute of Food Technologists®.)

There is a growing recognition of the role of arbuscular mycorrhizal fungi (AMF) in food security, specifically the potential for AMF to enhance the yield and mineral nutrition-including phosphorus, zinc (Zn), and iron (Fe)-of food crops. However, the bioavailability of Zn and Fe for humans in the grain of cereal crops can be overestimated by failing to consider the abundance of phytic acid (PA). This is because PA can chelate the micronutrients, making them difficult to absorb. In order to understand the effect of an AM fungus and soil Zn concentration on the productivity and nutritional quality of food parts, this study examined the growth and nutritional responses of durum wheat, with and without inoculation with Rhizophagus irregularis, at five soil Zn concentrations. Growth and nutrient responses of the plants to soil Zn amendment was stronger than responses to AMF. However, the protective effect of AMF under soil Zn toxicity conditions was observed as reduced Zn concentration in the mycorrhizal durum wheat grain at Zn50. Here, AMF inoculation increased the concentration of PA in durum wheat grain but had no effect on the concentration of Zn and Fe; this consequently reduced the predicted bioavailability of grain Zn and Fe, which could lead to a decrease in nutritional quality of the grain. This research suggests that in soil with low (available) phosphorus and Zn concentrations, AMF may reduce the food quality of durum wheat because of an increase in PA concentration, and thus, a decrease in the bioavailability of Zn and Fe.

Background: Hexaploid wheat is an important cereal crop that has been targeted to enhance grain micronutrient content including zinc (Zn) and iron (Fe). In this direction, modulating the expression of plant transporters involved in Fe and Zn homeostasis has proven to be one of the promising approaches. The present work was undertaken to identify wheat zinc-induced facilitator-like (ZIFL) family of transporters. The wheat ZIFL genes were characterized for their transcriptional expression response during micronutrient fluctuations and exposure to multiple heavy metals., Results: The genome-wide analyses resulted in identification of fifteen putative TaZIFL-like genes, which were distributed only on Chromosome 3, 4 and 5. Wheat ZIFL proteins subjected to the phylogenetic analysis showed the uniform distribution along with rice, Arabidopsis and maize. In-silico analysis of the promoters of the wheat ZIFL genes demonstrated the presence of multiple metal binding sites including those which are involved in Fe and heavy metal homeostasis. Quantitative real-time PCR analysis of wheat ZIFL genes suggested the differential regulation of the transcripts in both roots and shoots under Zn surplus and also during Fe deficiency. Specifically, in roots, TaZIFL2.3, TaZIFL4.1, TaZIFL4.2, TaZIFL5, TaZIFL6.1 and TaZIFL6.2 were significantly up-regulated by both Zn and Fe. This suggested that ZIFL could possibly be regulated by both the nutrient stress in a tissue specific manner. When exposed to heavy metals, TaZIFL4.2 and TaZIFL7.1 show significant up-regulation, whereas TaZIFL5 and TaZIFL6.2 remained almost unaffected., Conclusion: This is the first report for detailed analysis of wheat ZIFL genes. ZIFL genes also encode for transporter of mugineic acid (TOM) proteins, that are involved in the release of phytosiderophores to enhance Fe/Zn uptake. The detailed expression analysis suggests the varying expression patterns during development of wheat seedlings and also against abiotic/biotic stresses. Overall, this study will lay foundation to prioritize functional assessment of the candidate ZIFL as a putative TOM protein in wheat.

Helicobacter pylori infection is common in low-income countries. It has been associated with iron deficiency and reduced efficacy of iron supplementation. Whether H. pylori infection affects iron absorption from fortified and biofortified foods is unclear. Our objective was to assess whether asymptomatic H. pylori infection predicts dietary iron bioavailability in women and children, two main target groups of iron fortification programs. We did a pooled analysis of studies in women of reproductive age and preschool children that were conducted in Benin, Senegal and Haiti using stable iron isotope tracers to measure erythrocyte iron incorporation. We used mixed models to assess whether asymptomatic H. pylori infection predicted fractional iron absorption from ferrous sulfate, ferrous fumarate or NaFeEDTA, controlling for age, hemoglobin, iron status (serum ferritin), inflammation (C-reactive protein), and test meal. The analysis included 213 iron bioavailability measurements from 80 women and 235 measurements from 90 children; 51.3% of women and 54.4% of children were seropositive for H. pylori . In both women and children, hemoglobin (Hb), serum ferritin (SF), and C-reactive protein (CRP) did not differ between the seropositive and seronegative groups. Geometric mean (95% CI) fractional iron absorption (%), adjusted for SF, was 8.97% (7.64, 10.54) and 6.06% (4.80, 7.67) in H. pylori positive and negative women ( p = 0.274), and 9.02% (7.68, 10.59) and 7.44% (6.01, 9.20) in H. pylori positive and negative children ( p = 0.479). Our data suggest asymptomatic H. pylori infection does not predict fractional iron absorption from iron fortificants given to preschool children or young women in low-income settings.

The effect of soluble extracts with putative prebiotic ability extracted from various bean varieties on the intestinal brush border membrane (BBM) iron related proteins, and intestinal bacterial populations were evaluated using the Gallus gallus model and by the intra-amniotic administration procedure. Eight treatment groups [(non-injected; 18 MΩ H 2 O; 40 mg/mL Inulin; 50 mg/mL BRS Perola (carioca standard); 50 mg/mL BRS Cometa (carioca, Fe biofortified); 50 mg/mL BRS Esteio (black, standard); 50 mg/mL SMN 39 (black, Fe biofortified); 50 mg/mL BRS Artico (white, standard)] were utilized. Tested groups reduced the relative abundance of Clostridium and E. coli compared to the Inulin group (positive control) and they did not affect the relative abundance of Bifidobacterium and Lactobacillus compared to the negative control (18MΩ H 2 O). The relative expression of zinc transporter 1, ferroportin and amino peptidase were up-regulated in the BRS Cometa group (Fe-biofortified carioca beans). Results suggest that soluble extracts from carioca beans may improve the iron bioavailability by affecting intestinal bacterial populations, and BBM functionality., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Zinc (Zn) and iron (Fe) are essential micronutrients for plant growth. Thus, it is important to understand the mechanisms of uptake, transport and accumulation of these micronutrients in maize to improve crop nutritional quality. Members of the zinc-regulated transporters, iron-regulated transporter-like protein (ZIP) family are responsible for the uptake and transport of divalent metal ions in plant. Previously, we showed that ZmZIP5 functionally complemented the Zn uptake double mutant zrt1zrt2, Fe-uptake double mutant fet3fet4 in yeast. In our β-glucuronidase (GUS) assay, the germinated seeds, young sheaths, and stems of ZmZIP5-promoter-GUS transgenic plants were stained. We generated and compared two maize lines for this study: Ubi-ZmZIP5, in which ZmZIP5 was constitutively overexpressed, and ZmZIP5i, a RNAi line. At the seedling stage, high levels of Zn and Fe were found in the roots and shoots of Ubi-ZmZIP5 plants, whereas low levels were found in the ZmZIP5i plants. Zn and Fe contents decreased in the seeds of Ubi-ZmZIP5 plants and remained unchanged in the seeds of ZmZIP5i plants. The seeds of Leg-ZmZIP5 plants, in which ZmZIP5 overexpression is specific to the endosperm, had higher levels of Zn and Fe. Our results imply that ZmZIP5 may play a role in Zn and Fe uptake and root-to-shoot translocation. Endosperm-specific ZmZIP5 overexpression could be useful for Zn and Fe biofortification of cereal grains., (� The Author(s) 2019. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For permissions, please email: journals.permissions@oup.com.)

Iron (Fe) and zinc (Zn) deficiencies are wide spread in South Asia and Africa. Biofortification of food crops is a viable means of addressing micronutrient deficiencies. Lentil is an important pulse crop that provides affordable source of proteins, minerals, fibre and carbohydrates for micronutrient deficient countries. An association mapping (AM) panel of 96 diverse lentil genotypes from India and Mediterranean region was evaluated for three seasons and genotyped using 80 polymorphic simple-sequence repeat (SSR) markers for identification of the markers associated with grain Fe and Zn concentrations. A Bayesian model based clustering identified five subpopulations, adequately explaining the genetic structure of the AM panel. The linkage disequilibrium (LD) analysis using mixed linear model (MLM) identified two SSR markers, GLLC106 and GLLC108, associated with grain Fe concentration explaining 17% and 6% phenotypic variation, respectively and three SSR markers (PBALC 364, PBALC 92 and GLLC592) associated with grain Zn concentration, explaining 6%, 8% and 13% phenotypic variation, respectively. The identified SSRs exhibited consistent performance across three seasons and have potential for utilization in lentil molecular breeding programme.