Photosynthetic plasticity aggravates the susceptibility of magnesium-deficient leaf to high light in rapeseed plants: the importance of Rubisco and mesophyll conductance.

Plants grown under low magnesium (Mg) soils are highly susceptible to encountering light intensities that exceed the capacity of photosynthesis (A), leading to a depression of photosynthetic efficiency and eventually to photooxidation (i.e., leaf chlorosis). Yet, it remains unclear which processes play a key role in limiting the photosynthetic energy utilization of Mg-deficient leaves, and whether the plasticity of A in acclimation to irradiance could have cross-talk with Mg, hence accelerating or mitigating the photodamage. We investigated the light acclimation responses of rapeseed (Brassica napus) grown under low- and adequate-Mg conditions. Magnesium deficiency considerably decreased rapeseed growth and leaf A, to a greater extent under high than under low light, which is associated with higher level of superoxide anion radical and more severe leaf chlorosis. This difference was mainly attributable to a greater depression in dark reaction under high light, with a higher Rubisco fallover and a more limited mesophyll conductance to CO2 (gm ). Plants grown under high irradiance enhanced the content and activity of Rubisco and gm to optimally utilize more light energy absorbed. However, Mg deficiency could not fulfill the need to activate the higher level of Rubisco and Rubisco activase in leaves of high-light-grown plants, leading to lower Rubisco activation and carboxylation rate. Additionally, Mg-deficient leaves under high light invested more carbon per leaf area to construct a compact leaf structure with smaller intercellular airspaces, lower surface area of chloroplast exposed to intercellular airspaces, and CO2 diffusion conductance through cytosol. These caused a more severe decrease in within-leaf CO2 diffusion rate and substrate availability. Taken together, plant plasticity helps to improve photosynthetic energy utilization under high light but aggravates the photooxidative damage once the Mg nutrition becomes insufficient.

Micronutrient homeostasis in plants for more sustainable agriculture and healthier human nutrition.

The provision of sustainable, sufficient, and nutritious food to the growing population is a major challenge for agriculture and the plant research community. In this respect, the mineral micronutrient content of food crops deserves particular attention. Micronutrient deficiencies in cultivated soils and plants are a global problem that adversely affects crop production and plant nutritional value, as well as human health and well-being. In this review, we call for awareness of the importance and relevance of micronutrients in crop production and quality. We stress the need for better micronutrient nutrition in human populations, not only in developing but also in developed nations, and describe strategies to identify and characterize new varieties with high micronutrient content. Furthermore, we explain how adequate nutrition of plants with micronutrients impacts metabolic functions and the capacity of plants to express tolerance mechanisms against abiotic and biotic constraints. Finally, we provide a brief overview and a critical discussion on current knowledge, future challenges, and specific technological needs for research on plant micronutrient homeostasis. Research in this area is expected to foster the sustainable development of nutritious and healthy food crops for human consumption.

Complementary alternative medicine in rheumatic diseases: Causes, choices, and outcomes according to patients.

OBJECTIVE: The knowledge of physicians about complementary and alternative medicine (CAM) applications is limited. However, especially in chronic diseases, patients and their relatives can often refer to CAM applications. Rheumatic diseases are chronic in nature presenting with a wide clinical spectrum. Despite developing treatment options, achieving treatment goals may not always be possible. For this reason, patients seek different treatment and use traditional and complementary medicine. The aim of this study was to investigate causes, consequences, and the frequency of applying to CAM in rheumatic diseases. METHODS: Ninety-five patients admitted to the rheumatology outpatient clinic were enrolled in the study. Health assessment questionnaire and short-form-36 were used to determine the quality of life of patients. Anxiety and depression symptoms were screened by the Hospital Anxiety and Depression scale, a questionnaire that was filled-in by the patients themselves. Also, patients were questioned about their place of residence, level of education, diagnosis, CAM modality types, application reasons, and outcomes. Chi-square test was used to analyze categorical data. Parametric data were analyzed using Student t-test, and nonparametric data were analyzed using Mann-Whitney U test. RESULTS: Thirty-two of our patients had applied to CAM modalities (phytotherapy [34.45%], cupping therapy [21.8%], acupuncture [12.5%], hirudotherapy [12.5%], food supplement [12.5%], and ozone treatment [6.25%]). Only 31.3% of the patients informed their doctors about CAM applications. 47.8% of fibromyalgia patients and 29.2% of patients with inflammatory rheumatic diseases had applied to CAM. Gender, working status, income level, smoking, and alcohol habits were not associated with the application to CAM. However, none of the residents of the village, 14.3% of the residents of the district center, and 41.1% of the residents of the city center had applied to CAM modality. The rate of applying to CAM was 18.2% for those who cannot read and write. The application ratio of CAM is over 40% among secondary school, high school, and university graduates. CONCLUSION: Among patients with rheumatic diseases, application to CAM is quite common. Very few patients inform their physicians about applying to CAM. Contrary to what is presumed, the rate of applying CAM applications is lower among those living in rural areas and with low education levels.

Feeding Zinc-Biofortified Wheat Improves Performance, Nutrient Digestibility, and Concentrations of Blood and Tissue Minerals in Quails.

The present study aimed to investigate the effects of feeding zinc (Zn)-biofortified wheat on performance, digestibility, and concentrations of minerals in quails. Zinc biofortification of wheat has been realized in the field by ergonomically applying Zn to foliar two and three times, which increased grain Zn from 18 mg/kg (control) to 34 and 64 mg/kg. A total of 180 quails were divided into six groups, each containing 30 birds, and fed diets containing wheat grain with either 18, 34, or 64 mg/kg with or without zinc picolinate (ZnPic) supplementation. Bodyweight, feed intake, feed efficiency, and cold carcass weights were greater (P = 0.0001) when the quails were fed a diet containing the biofortified wheat-containing 64 mg Zn/kg. Nitrogen, ash, Ca, P, Zn, Cu, and Fe retentions were greater with the Zn-biofortified wheat-containing 64 mg Zn/kg (P ≤ 0.026). The nutrient excretions were low with feeding a diet containing biofortified wheat-containing 64 mg Zn/kg (P ≤ 0.023). Serum, liver, and heart Zn concentrations increased with feeding biofortified wheat-containing 64 mg Zn/kg (P ≤ 0.002). Thigh meat Fe concentrations increased with increasing Zn concentrations of the wheat samples used (P = 0.0001), whereas the liver Cu concentrations decreased with feeding the wheat-containing 64 mg Zn/kg (P = 0.004). The Zn-biofortified wheat-containing greater Zn concentrations, particularly 64 mg Zn/kg, is a good replacement for corn in the poultry diet as long as its availability and low cost for better performance, greater digestibility, and elevated tissue Zn and Fe concentrations.

Biofortification of Silage Maize with Zinc, Iron and Selenium as Affected by Nitrogen Fertilization.

Agronomic biofortification is one of the main strategies for alleviation of micronutrient deficiencies in human populations and promoting sustainable production of food and feed. The aim of this study was to investigate the effect of nitrogen (N)fertilization on biofortification of maize crop (Zea mays L.) with zinc (Zn), iron (Fe) and selenium (Se) grown on a micronutrient deficient soil under greenhouse conditions. Factorial design experiment was set under greenhouse conditions. The experiment consisted of two levels of each N, Zn, Fe and Se. The levels for N were 125 and 250 mg N kg-1 soil; Zn were 1 and 5 mg Zn kg-1 soil; levels of Fe were 0 and 10 mg Fe kg-1 soil; levels of Se were 0 and 0.02 mg Se kg-1 soil. An additional experiment was also conducted to study the effect of the Zn form applied as a ZnO or ZnSO4 on shoot growth, shoot Zn concentration and total shoot Zn uptake per plant. Shoot Zn concentrations increased by increasing soil Zn application both with ZnSO4 and ZnO treatments, but the shoot Zn concentration and total Zn uptake were much greater with ZnSO4 than the ZnO application. Under given experimental conditions, increasing soil N supply improved shoot N concentration; but had little effect on shoot dry matter production. The concentrations of Zn and Fe in shoots were significantly increased by increasing N application. In case of total uptake of Zn and Fe, the positive effect of N nutrition was more pronounced. Although Se soil treatment had significant effect, N application showed no effect on Se concentration and accumulation in maize shoots. The obtained results show that N fertilization is an effective tool in improving the Zn and Fe status of silage maize and contribute to the better-quality feed.

Feed Composition Differences Resulting from Organic and Conventional Farming Practices Affect Physiological Parameters in Wistar Rats-Results from a Factorial, Two-Generation Dietary Intervention Trial.

Recent human cohort studies reported positive associations between organic food consumption and a lower incidence of obesity, cancer, and several other diseases. However, there are very few animal and human dietary intervention studies that provide supporting evidence or a mechanistic understanding of these associations. Here we report results from a two-generation, dietary intervention study with male Wistar rats to identify the effects of feeds made from organic and conventional crops on growth, hormonal, and immune system parameters that are known to affect the risk of a number of chronic, non-communicable diseases in animals and humans. A 2 × 2 factorial design was used to separate the effects of contrasting crop protection methods (use or non-use of synthetic chemical pesticides) and fertilizers (mineral nitrogen, phosphorus and potassium (NPK) fertilizers vs. manure use) applied in conventional and organic crop production. Conventional, pesticide-based crop protection resulted in significantly lower fiber, polyphenol, flavonoid, and lutein, but higher lipid, aldicarb, and diquat concentrations in animal feeds. Conventional, mineral NPK-based fertilization resulted in significantly lower polyphenol, but higher cadmium and protein concentrations in feeds. Feed composition differences resulting from the use of pesticides and/or mineral NPK-fertilizer had a significant effect on feed intake, weight gain, plasma hormone, and immunoglobulin concentrations, and lymphocyte proliferation in both generations of rats and in the second generation also on the body weight at weaning. Results suggest that relatively small changes in dietary intakes of (a) protein, lipids, and fiber, (b) toxic and/or endocrine-disrupting pesticides and metals, and (c) polyphenols and other antioxidants (resulting from pesticide and/or mineral NPK-fertilizer use) had complex and often interactive effects on endocrine, immune systems and growth parameters in rats. However, the physiological responses to contrasting feed composition/intake profiles differed substantially between the first and second generations of rats. This may indicate epigenetic programming and/or the generation of "adaptive" phenotypes and should be investigated further.

Optimization of Potassium Supply under Osmotic Stress Mitigates Oxidative Damage in Barley.

Potassium (K) is the most abundant cation in plants, playing an important role in osmoregulation. Little is known about the effect of genotypic variation in the tolerance to osmotic stress under different K treatments in barley. In this study, we measured the interactive effects of osmotic stress and K supply on growth and stress responses of two barley cultivars (Hordeum vulgare L.) and monitored reactive oxygen species (ROS) along with enzymatic antioxidant activity and their respective gene expression level. The selected cultivars (cv. Milford and cv. Sahin-91Sahin-91) were exposed to osmotic stress (-0.7 MPa) induced by polyethylene glycol 6000 (PEG) under low (0.04 mM) and adequate (0.8 mM) K levels in the nutrient solution. Leaf samples were collected and analyzed for levels of K, ROS, kinetic activity of antioxidants enzymes and expression levels of respective genes during the stress period. The results showed that optimal K supply under osmotic stress significantly decreases ROS production and adjusts antioxidant activity, leading to the reduction of oxidative stress in the studied plants. The cultivar Milford had a lower ROS level and a better tolerance to stress compared to the cultivar Sahin-91. We conclude that optimized K supply is of great importance in mitigating ROS-related damage induced by osmotic stress, specifically in drought-sensitive barley cultivars.

Simultaneous Biofortification of Rice With Zinc, Iodine, Iron and Selenium Through Foliar Treatment of a Micronutrient Cocktail in Five Countries.

Widespread malnutrition of zinc (Zn), iodine (I), iron (Fe) and selenium (Se), known as hidden hunger, represents a predominant cause of several health complications in human populations where rice (Oryza sativa L.) is the major staple food. Therefore, increasing concentrations of these micronutrients in rice grain represents a sustainable solution to hidden hunger. This study aimed at enhancing concentration of Zn, I, Fe and Se in rice grains by agronomic biofortification. We evaluated effects of foliar application of Zn, I, Fe and Se on grain yield and grain concentration of these micronutrients in rice grown at 21 field sites during 2015 to 2017 in Brazil, China, India, Pakistan and Thailand. Experimental treatments were: (i) local control (LC); (ii) foliar Zn; (iii) foliar I; and (iv) foliar micronutrient cocktail (i.e., Zn + I + Fe + Se). Foliar-applied Zn, I, Fe or Se did not affect rice grain yield. However, brown rice Zn increased with foliar Zn and micronutrient cocktail treatments at all except three field sites. On average, brown rice Zn increased from 21.4 mg kg-1 to 28.1 mg kg-1 with the application of Zn alone and to 26.8 mg kg-1 with the micronutrient cocktail solution. Brown rice I showed particular enhancements and increased from 11 µg kg-1 to 204 µg kg-1 with the application of I alone and to 181 µg kg-1 with the cocktail. Grain Se also responded very positively to foliar spray of micronutrients and increased from 95 to 380 µg kg-1. By contrast, grain Fe was increased by the same cocktail spray at only two sites. There was no relationship between soil extractable concentrations of these micronutrients with their grain concentrations. The results demonstrate that irrespective of the rice cultivars used and the diverse soil conditions existing in five major rice-producing countries, the foliar application of the micronutrient cocktail solution was highly effective in increasing grain Zn, I and Se. Adoption of this agronomic practice in the target countries would contribute significantly to the daily micronutrient intake and alleviation of micronutrient malnutrition in human populations.

Magnesium Deficiency Reduced the Yield and Seed Germination in Wax Gourd by Affecting the Carbohydrate Translocation.

Magnesium (Mg) is a particular mineral nutrient greatly affecting the size and activity of sink organs. Wax gourd crop with its fruits having fresh weight up to 20-50 kg per single fruit serves as an excellent experimental plant species for better understanding the role of varied Mg nutrition in sink strength and yield formation. This study aimed to investigate the effects of Mg deficiency on fruit yield and seed vigor in wax gourd grown under field conditions. Plants were grown under field conditions until maturity with increasing soil Mg applications. At the beginning of fruit formation, leaves were used to analyze concentrations of sucrose, starch and Mg as well as phloem export of sucrose. At maturity, fruit yield was determined and the seeds collected were used in germination studies and starch analysis. Low Mg supply resulted in a significant impairment in fruit fresh yield, which was closely associated with higher accumulation of starch and sucrose in source leaves and lower amount of sucrose in phloem exudate. Seeds obtained from Mg deficiency plants exhibited lower amount of starch and substantial reduction in both germination capacity and seedling establishment when compared to the seeds from the Mg adequate plants. Our study revealed that magnesium deficiency significantly diminished fruit yield of field-grown wax gourd, most probably by limiting the carbohydrate transport from source organs to developing fruit. Ensuring sufficient Mg supply to plant species with high sink size such as wax gourd, during the reproductive growth stage, is a critical factor for achieving higher fruit yield formation and also better vigor of next-generation seeds.

Fate and Bioaccessibility of Iodine in Food Prepared from Agronomically Biofortified Wheat and Rice and Impact of Cofertilization with Zinc and Selenium.

Enrichment of food crops with iodine is an option to alleviate dietary deficiencies. Therefore, foliar iodine fertilizer was applied on wheat and rice, in the presence and absence of the other micronutrients zinc and selenium. This treatment increased the concentration of iodine, as well as zinc and selenium, in the staple grains. Subsequently, potential iodine losses during preparation of foodstuffs with the enriched grains were studied. Oven-heating did not affect the iodine content in bread. Extraction of bran from flour lowered the iodine in white bread compared to wholegrain bread, but it was still markedly higher compared to the control. During subsequent in vitro gastrointestinal digestion, a higher percentage of iodine was released from foods based on extracted flour (82-92%) compared to wholegrain foods (50-76%). The foliar fertilization of wheat was found to be adequate to alleviate iodine deficiency in a population with a moderate to high intake of bread.

Variation in phosphorus and sulfur content shapes the genetic architecture and phenotypic associations within the wheat grain ionome.

Dissection of the genetic basis of wheat ionome is crucial for understanding the physiological and biochemical processes underlying mineral accumulation in seeds, as well as for efficient crop breeding. Most of the elements essential for plants are metals stored in seeds as chelate complexes with phytic acid or sulfur-containing compounds. We assume that the involvement of phosphorus and sulfur in metal chelation is the reason for strong phenotypic correlations within ionome. Adjustment of element concentrations for the effect of variation in phosphorus and sulfur seed content resulted in drastic change of phenotypic correlations between the elements. The genetic architecture of wheat grain ionome was characterized by quantitative trait loci (QTL) analysis using a cross between durum and wild emmer wheat. QTL analysis of the adjusted traits and two-trait analysis of the initial traits paired with either P or S considerably improved QTL detection power and accuracy, resulting in the identification of 105 QTLs and 617 QTL effects for 11 elements. Candidate gene search revealed some potential functional associations between QTLs and corresponding genes within their intervals. Thus, we have shown that accounting for variation in P and S is crucial for understanding of the physiological and genetic regulation of mineral composition of wheat grain ionome and can be implemented for other plants.

Simultaneous Biofortification of Wheat with Zinc, Iodine, Selenium, and Iron through Foliar Treatment of a Micronutrient Cocktail in Six Countries.

Field experiments were conducted on wheat to study the effects of foliar-applied iodine(I) alone, Zn (zinc) alone, and a micronutrient cocktail solution containing I, Zn, Se (selenium), and Fe (iron) on grain yield and grain concentrations of micronutrients. Plants were grown over 2 years in China, India, Mexico, Pakistan, South Africa, and Turkey. Grain-Zn was increased from 28.6 mg kg-1 to 46.0 mg-1 kg with Zn-spray and 47.1 mg-1 kg with micronutrient cocktail spray. Foliar-applied I and micronutrient cocktail increased grain I from 24 µg kg-1 to 361 µg kg-1 and 249 µg kg-1, respectively. Micronutrient cocktail also increased grain-Se from 90 µg kg-1 to 338 µg kg-1 in all countries. Average increase in grain-Fe by micronutrient cocktail solution was about 12%. The results obtained demonstrated that foliar application of a cocktail micronutrient solution represents an effective strategy to biofortify wheat simultaneously with Zn, I, Se and partly with Fe without yield trade-off in wheat.

Zinc Absorption From Agronomically Biofortified Wheat Is Similar to Post-Harvest Fortified Wheat and Is a Substantial Source of Bioavailable Zinc in Humans.

BACKGROUND: Limited data exist on human zinc absorption from wheat biofortified via foliar (FBW) or root (hydroponically fortified wheat, HBW) zinc application. Stable isotope labels added at point of consumption (extrinsic labeling) might not reflect absorption from native zinc obtained by intrinsic labeling. OBJECTIVES: We measured fractional and total zinc absorption (FAZ, TAZ) in FBW and HBW wheat, compared with control wheat (CW) and fortified wheat (FW). The effect of labeling method was assessed in HBW (study 1), and the effect of milling extraction rate (EXR, 80% and 100%) in FBW (studies 2 and 3). METHODS: Generally healthy adults (n = 71, age: 18-45 y, body mass index: 18.5-25 kg/m2) were allocated to 1 of the studies, in which they served as their own controls. In study 1, men and women consumed wheat porridges colabeled intrinsically and extrinsically with 67Zn and 70Zn. In studies 2 and 3, women consumed wheat flatbreads (chapatis) labeled extrinsically. Zinc absorption was measured with the oral to intravenous tracer ratio method with a 4-wk wash-out period between meals. Data were analyzed with linear mixed models. RESULTS: In study 1 there were no differences in zinc absorption from extrinsic versus intrinsic labels in either FW or HBW. Similarly, FAZ and TAZ from FW and HBW did not differ. TAZ was 70-76% higher in FW and HBW compared with CW (P < 0.01). In studies 2 and 3, TAZ from FW and FBW did not differ but was 20-48% higher compared with CW (P < 0.001). Extraction rate had no effect on TAZ. CONCLUSIONS: Colabeling demonstrates that extrinsic zinc isotopic labels can be used to accurately quantify zinc absorption from wheat in humans. Biofortification through foliar zinc application, root zinc application, or fortification provides higher TAZ compared with unfortified wheat. In biofortified wheat, extraction rate (100-80%) has a limited impact on total zinc absorption. These studies were registered on clinicaltrials.gov (NCT01775319).

Effect of zinc-biofortified seeds on grain yield of wheat, rice, and common bean grown in six countries.

Seeds enriched with zinc (Zn) are ususally associated with better germination, more vigorous seedlings and higher yields. However, agronomic benefits of high-Zn seeds were not studied under diverse agro-climatic field conditions. This study investigated effects of low-Zn and high- Zn seeds (biofortified by foliar Zn fertilization of maternal plants under field conditions) of wheat (Tritcum aestivum L.), rice (Oryza sativa L.), and common bean (Phaseolus vulgaris L.) on seedling density, grain yield and grain Zn concentration in 31 field locations over two years in six countries. Experimental treatments were: (1) low-Zn seeds and no soil Zn fertilization (control treatment), (2) low-Zn seeds + soil Zn fertilization, and (3) Zn-biofortified seeds and no soil Zn fertilization. The wheat experiments were established in China, India, Pakistan, and Zambia, the rice experiments in China, India and Thailand, and the common bean experiment in Brazil. When compared to the control treatment, soil Zn fertilization increased wheat grain yield in all six locations in India, two locations in Pakistan and one location in China. Zinc-biofortified seeds also increased wheat grain yield in all four locations in Pakistan and four locations in India compared to the control treatment. Across all countries over 2 years, Zn-biofortified wheat seeds increased plant population by 26.8% and grain yield by 5.37%. In rice, soil Zn fertilization increased paddy yield in all four locations in India and one location in Thailand. Across all countries, paddy yield increase was 8.2% by soil Zn fertilization and 5.3% by Zn-biofortified seeds when compared to the control treatment. In common bean, soil Zn application as well as Znbiofortified seed increased grain yield in one location in Brazil. Effects of soil Zn fertilization and high-Zn seed on grain Zn density were generally low. This study, at 31 field locations in six countries over two years, revealed that the seeds biofortfied with Zn enhanced crop productivity at many locations with different soil and environmental conditions. As high-Zn grains are a by-product of Zn biofortification, use of Zn-enriched grains as seed in the next cropping season can contribute to enhance crop productivity in a cost-effective manner.

Effects of Agronomic Management and Climate on Leaf Phenolic Profiles, Disease Severity, and Grain Yield in Organic and Conventional Wheat Production Systems.

Agricultural intensification over the last 40 years has increased cereal yields, but there is very limited information on the effects of intensification practices (e.g., nondiverse rotations, mineral NPK fertilizer, and pesticides) on crop health and quality. Results from the study reported here suggest that the use of mineral NPK fertilizers reduces phenolic acid and flavonoid concentrations in leaves and increases the susceptibility of wheat to lodging and powdery mildew, when compared to composted FYM inputs. In contrast, the use of herbicides, fungicides, and growth regulators reduces lodging and foliar disease severity but had no effect on phenolic acid and flavonoid concentrations. The use of composted FYM inputs also resulted in a significant grain yield reduction and not substantially reduced the severity of opportunistic pathogens such as Septoria, which remain a major yield limiting factor unless fungicides are used and/or more Septoria resistant varieties become available.

Quantitative trait loci associated with soybean seed weight and composition under different phosphorus levels.

Seed size and composition are important traits in food crops and can be affected by nutrient availability in the soil. Phosphorus (P) is a non-renewable, essential macronutrient, and P deficiency limits soybean (Glycine max) yield and quality. To investigate the associations of seed traits in low- and high-P environments, soybean recombinant inbred lines (RILs) from a cross of cultivars Fiskeby III and Mandarin (Ottawa) were grown under contrasting P availability environments. Traits including individual seed weight, seed number, and intact mature pod weight were significantly affected by soil P levels and showed transgressive segregation among the RILs. Surprisingly, P treatments did not affect seed composition or weight, suggesting that soybean maintains sufficient P in seeds even in low-P soil. Quantitative trait loci (QTLs) were detected for seed weight, intact pods, seed volume, and seed protein, with five significant QTLs identified in low-P environments and one significant QTL found in the optimal-P environment. Broad-sense heritability estimates were 0.78 (individual seed weight), 0.90 (seed protein), 0.34 (seed oil), and 0.98 (seed number). The QTLs identified under low P point to genetic regions that may be useful to improve soybean performance under limiting P conditions.

Harvesting more grain zinc of wheat for human health.

Increasing grain zinc (Zn) concentration of cereals for minimizing Zn malnutrition in two billion people represents an important global humanitarian challenge. Grain Zn in field-grown wheat at the global scale ranges from 20.4 to 30.5 mg kg-1, showing a solid gap to the biofortification target for human health (40 mg kg-1). Through a group of field experiments, we found that the low grain Zn was not closely linked to historical replacements of varieties during the Green Revolution, but greatly aggravated by phosphorus (P) overuse or insufficient nitrogen (N) application. We also conducted a total of 320-pair plots field experiments and found an average increase of 10.5 mg kg-1 by foliar Zn application. We conclude that an integrated strategy, including not only Zn-responsive genotypes, but of a similar importance, Zn application and field N and P management, are required to harvest more grain Zn and meanwhile ensure better yield in wheat-dominant areas.

Molecular speciation and tissue compartmentation of zinc in durum wheat grains with contrasting nutritional status.

Low concentration of zinc (Zn) in the endosperm of cereals is a major factor contributing to Zn deficiency in human populations. We have investigated how combined Zn and nitrogen (N) fertilization affects the speciation and localization of Zn in durum wheat (Triticum durum). Zn-binding proteins were analysed with liquid chromatography ICP-MS and Orbitrap MS(2) , respectively. Laser ablation ICP-MS with simultaneous Zn, sulphur (S) and phosphorus (P) detection was used for bioimaging of Zn and its potential ligands. Increasing the Zn and N supply had a major impact on the Zn concentration in the endosperm, reaching concentrations higher than current breeding targets. The S concentration also increased, but S was only partly co-localized with Zn. The mutual Zn and S enrichment was reflected in substantially more Zn bound to small cysteine-rich proteins (apparent size 10-30 kDa), whereas the response of larger proteins (apparent size > 50 kDa) was only modest. Most of the Zn-responsive proteins were associated with redox- and stress-related processes. This study offers a methodological platform to deepen the understanding of processes behind endosperm Zn enrichment. Novel information is provided on how the localization and speciation of Zn is modified during Zn biofortification of grains.