Zinc (Zn) deficiency associated with low dietary intake is a well-documented public health problem, resulting in serious health and socioeconomic problems. Field experiments were conducted with wheat to test the role of both soil and foliar application of ZnSO4 in Zn concentration of whole grain and grain fractions (e.g., bran, embryo and endosperm) in 3 locations. Foliar application of ZnSO4 was realized at different growth stages (e.g., stem elongation, boot, milk, dough stages) to study the effect of timing of foliar Zn application on grain Zn concentration. The rate of foliar Zn application at each growth stage was 4 kg of ZnSO4·7H2O ha(-1). Laser ablation (LA)-ICP-MS was used to follow the localization of Zn within grain. Soil Zn application at a rate of 50 kg of ZnSO4·7H2O ha(-1) was effective in increasing grain Zn concentration in the Zn-deficient location, but not in the locations without soil Zn deficiency. In all locations, foliar application of Zn significantly increased Zn concentration in whole grain and in each grain fraction, particularly in the case of high soil N fertilization. In Zn-deficient location, grain Zn concentration increased from 11 mg kg(-1) to 22 mg kg(-1) with foliar Zn application and to 27 mg kg(-1) with a combined application of ZnSO4 to soil and foliar. In locations without soil Zn deficiency, combination of high N application with two times foliar Zn application (e.g., at the booting and milk stages) increased grain Zn concentration, on average, from 28 mg kg(-1) to 58 mg kg(-1). Both ICP-OES and LA-ICP-MS data showed that the increase in Zn concentration of whole grain and grain fractions was pronounced when Zn was sprayed at the late growth stage (e.g., milk and dough). LA-ICP-MS data also indicated that Zn was transported into endosperm through the crease phloem. To our knowledge, this is the first study to show that the timing of foliar Zn application is of great importance in increasing grain Zn in wheat, especially in the endosperm part that is the predominant grain fraction consumed in many countries. Providing a large pool of Zn in vegetative tissues during the grain filling (e.g., via foliar Zn spray) is an important practice to increase grain Zn and contribute to human nutrition.

In a field experiment, the effect of foliar Zn applications on the concentration of Zn in seeds of a bread wheat cultivar (Triticum aestivum L. cv. Balatilla) was studied during different stages of seed development. In addition, a staining method using dithizone (DTZ: diphenyl thiocarbazone) was applied to (1) study the localization of Zn in seeds, (2) follow the remobilization of Zn during germination, and (3) develop a rapid visual Zn screening method for seed and flour samples. In all seed development stages, foliar Zn treatments were effective in increasing seed Zn concentration. The highest Zn concentration in the seeds was found in the first stage of seed development (around the early milk stage); after this, seed Zn concentration gradually decreased until maturity. When reacting with Zn, DTZ forms a red-colored complex. The DTZ staining of seed samples revealed that Zn is predominantly located in the embryo and aleurone parts of the seeds. After 36 h of germination, the coleoptile and roots that emerged from seeds showed very intensive red color formation and had Zn concentrations up to 200 mg kg ÿ1 , indicating a substantial remobilization of Zn from seed pools into the developing roots (radicle) and coleoptile. The DTZ staining method seems to be useful in ranking flour samples for their Zn concentrations. There was a close relationship between the seed Zn concentrations and spectral absorbance of the methanol extracts of the flour samples stained with DTZ. The results suggest that (1) accumulation of Zn in seeds is particularly high during early seed development, (2) Zn is concentrated in the embryo and aleurone parts, and (3) the DTZ staining method can be used as a rapid, semiquantitative method to estimate Zn concentrations of flour and seed samples and to screen genotypes for their Zn concentrations in seeds.

Mineral nutrient malnutrition, and particularly deficiency in zinc and iron, afflicts over 3 billion people worldwide. Wild emmer wheat, Triticum turgidum ssp. dicoccoides, genepool harbors a rich allelic repertoire for mineral nutrients in the grain. The genetic and physiological basis of grain protein, micronutrients (zinc, iron, copper and manganese) and macronutrients (calcium, magnesium, potassium, phosphorus and sulfur) concentration was studied in tetraploid wheat population of 152 recombinant inbred lines (RILs), derived from a cross between durum wheat (cv. Langdon) and wild emmer (accession G18-16). Wide genetic variation was found among the RILs for all grain minerals, with considerable transgressive effect. A total of 82 QTLs were mapped for 10 minerals with LOD score range of 3.2-16.7. Most QTLs were in favor of the wild allele (50 QTLs). Fourteen pairs of QTLs for the same trait were mapped to seemingly homoeologous positions, reflecting synteny between the A and B genomes. Significant positive correlation was found between grain protein concentration (GPC), Zn, Fe and Cu, which was supported by significant overlap between the respective QTLs, suggesting common physiological and/or genetic factors controlling the concentrations of these mineral nutrients. Few genomic regions (chromosomes 2A, 5A, 6B and 7A) were found to harbor clusters of QTLs for GPC and other nutrients. These identified QTLs may facilitate the use of wild alleles for improving grain nutritional quality of elite wheat cultivars, especially in terms of protein, Zn and Fe.

Sixty-six spring and winter common wheat genotypes from Central Asian breeding programs were evaluated for grain concentrations of iron (Fe) and zinc (Zn). Iron showed large variation among genotypes, ranging from 25 mg kg -1 to 56 mg kg -1 (mean 38 mg kg -1 ). Similarly, Zn concentration varied among genotypes, ranging between 20 mg kg -1 and 39 mg kg -1 (mean 28 mg kg -1 ). Spring wheat cultivars possessed higher Fe-grain concentrations than winter wheats. By contrast, winter wheats showed higher Zn-grain concentrations than spring genotypes.

Increasing zinc (Zn) concentration of cereal grains is a global challenge to alleviate Zn deficiency‐related health problems in humans caused by low dietary Zn intake. This study investigated the effects of soil‐ and foliar‐applied nitrogen (N) and Zn fertilizers on grain Zn accumulation of durum wheat (Triticum durum) grown on a Zn‐deficient soil. In addition, localization of Zn and protein within durum wheat grain was studied by using Bradford reagent for protein and dithizone (diphenyl thiocarbazone) for Zn. Grain Zn concentration was greatly enhanced by soil or foliar applications of Zn. When Zn supply was adequately high, both soil and foliar N applications improved grain Zn concentration. Consequently, there was a significant positive correlation between grain concentrations of Zn and N, when Zn supply was not limiting. Protein and Zn staining studies showed co‐localization of Zn and protein within grain, particularly in the embryo and aleurone. Results indicate that N and Zn fertilization have a synergistic effect on grain Zn concentration. Possibly, increasing N supply contributes to grain Zn concentration by affecting the levels of Zn‐chelating nitrogenous compounds or the abundance of Zn transporters. Our results suggest that nitrogen management can be an effective agronomic tool to improve grain Zn concentration.

Micronutrient malnutrition afflicts over three billion people worldwide and the numbers are continuously increasing. Developing genetically micronutrientenriched cereals, which are the predominant source of human dietary, is essential to alleviate malnutrition worldwide. Wheat chromosome 6B derived from wild emmer wheat [Triticum turgidum ssp. dicoccoides (Kö rn.) Thell] was previously reported to be associated with high Zn concentration in the grain. In the present study, recombinant chromosome substitution lines (RSLs), previously constructed for genetic and physical maps of Gpc-B1 (a 250-kb locus affecting grain protein concentration), were used to identify the effects of the Gpc-B1 locus on grain micronutrient concentrations. RSLs carrying the Gpc-B1 allele of T. dicoccoides accumulated on average 12% higher concentration of Zn, 18% higher concentration of Fe, 29% higher concentration of Mn and 38% higher concentration of protein in the grain as compared with RSLs carrying the allele from cultivated wheat (Triticum durum). Furthermore, the high grain Zn, Fe and Mn concentrations were consistently expressed in five different environments with an absence of genotype by environment interaction. The results obtained in the present study also confirmed the previously reported effect of the wild-type allele of Gpc-B1 on earlier senescence of flag leaves. We suggest that the Gpc-B1 locus is involved in more efficient remobilization of protein, zinc, iron and manganese from leaves to the grains, in addition to its effect on earlier senescence of the green tissues.

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