Biofortification strategies to increase grain zinc and iron concentrations in wheat

Micronutrient deficiencies, especially those arising from zinc (Zn) and iron (Fe), pose serious human health problems for more than 2 billion people worldwide. Wheat is a major source of dietary energy and protein for the world's growing population, and its potential to assist in reducing micronutrient-related malnutrition can be enhanced via integration of agronomic fertilization practices and delivery of genetically-manipulated, micronutrient rich wheat varieties. Targeted breeding for these biofortified varieties was initiated by exploiting available genetic diversity for Zn and Fe from wild relatives of cultivated wheat and synthetic hexaploid progenitors. The proof-of-concept results from the performance of competitive biofortified wheat lines showed good adaptation in target environments without compromising essential core agronomic traits. Agronomic biofortification through fertilizer approaches could complement the existing breeding approach; for instance, foliar application of Zn fertilizer can increase grain Zn above the breeding target set by nutritionists. This review synthesizes the progress made in genetic and agronomic biofortification strategies for Zn and Fe enrichment of wheat.     

小麦籽粒微量元素含量的研究进展

以禾谷类作物为主食引起的人体摄取必需微量元素(特别是Fe和Zn)不足,已对现代社会和谐发展造成沉重的经济负担和安全隐患。小麦(Triticum aestivumL.)是中国和全球大多数人主要的食物和矿质元素来源。小麦籽粒中Zn、Fe含量普遍较低,已引起国内外学者们的高度关注。本文综述了小麦籽粒微量元素含量器官间、基因型间的差异及其影响因素和相关性状研究进展,介绍了小麦籽粒吸收和富集微量元素的生理与遗传基础,展望了提高小麦籽粒微量元素的研究内容和方向。     

Protein,calcium, iron,and amino acid content of selected wild and domesticated cultivars of finger millet

Two wild and eight domesticated cultivars of finger millet were analyzed to determine their proximate composition and calcium, iron, and amino acid content. Wide variations were observed in the protein (mean values ranged from 7.5 to 11.7%), calcium (376 to 515 mg/100 g), and iron (3.7 to 6.8 mg/100 g) content of the wild and domesticated cultivars. A wild progenitor of finger millet, Ecoracana subsp.africana was significantly higher in protein than four of the six domesticated accessions analyzed. The calcium and iron content of the wild progenitor was also significantly greater than that of two domesticated cultivars. The wild species was also found to be higher in lysine and five other essential amino acids. These results indicate that the nutritional value of finger millet may be significantly improved by selective crossbreeding of the cereal's wild and domesticated cultivars.     

不同基因型小麦籽粒、面粉和麸皮中Ca和Zn含量的差异

为了筛选富含人体必需营养元素钙（Ca）和锌（Zn）的小麦品种,改善食品营养结构,以来自长江中下游地区的推广品种和部分国内外引进的小麦品种（共112个）为材料,进行了籽粒Ca和Zn含量的测定和分析,通过聚类分析选出其中39个代表性品种,对其麸皮和面粉中的Ca和Zn含量进行了进一步分析。结果表明,不同基因型品种籽粒中Ca和Zn含量的变异范围较宽,品种间存在极显著差异,面粉、麸皮和籽粒中均存在含量较高的品种。Ca和Zn含量均表现为麸皮中最高,变异幅度最宽,籽粒中次之,面粉中最低且变异幅度也较小,但“面粉麸皮籽粒”中营养元素的含量关系比较复杂,并不总是呈显著正相关。品种NP164、青紫1号和荆州D402等可作为提高当地小麦面粉Ca和Zn含量的重要种质资源加以应用。     

碳氮供给对小麦离体穗培养粒重、微量元素及蛋白质含量的影响

为了解碳氮供给与小麦粒重、Fe、Zn、Mn、Cu等微量元素以及蛋白质含量的关系,在离体穗培养条件下研究了灌浆初期和灌浆中期不同浓度C(蔗糖)、N(硝酸铵)供给对小麦粒重、微量元素(Fe、Zn、Mn、Cu)和蛋白质含量的影响.结果表明,随着培养基糖浓度的增大,粒重和Fe、Mn含量都表现为先升高后降低的趋势,在4%糖浓度时,均达到最高值;籽粒Zn、Cu和蛋白质含量表现为随糖浓度增大而持续降低.随着培养基N浓度的增加,粒重和Fe、Zn、Mn、Cu含量多表现为先升高后降低的趋势,且多在N浓度为0.07%时达到最高值,尤以灌浆中期开始的培养表现明显;籽粒蛋白质含量表现为随N浓度增大而持续增加.由此可见,外源C、N供给对粒重、微量元素和蛋白质含量有明显的调控效应.一定的糖供给可同时提高粒重和Fe、Mn含量,一定的N供给可同时提高粒重及微量元素和蛋白质含量.     

Genetic variability and diversity for protein and calcium contents in finger millet (Eleusine coracana (L.) Gaertn) in relation to grain color

Analysis of 36 genotypes of finger millet ( Eleusine coracana (L.) Gaertn) with varying seed colors revealed a wide range of protein and calcium contents. White seeded genotypes had higher protein contents, while brown seeded types had a wide range of values. The brown seeded genotype GE 2500 had the highest protein content. Although protein content had significant negative association with calcium content, white seeded types had moderate levels of calcium. The genotypic coefficients of variability were moderate and high for protein and calcium, respectively. High heritability coupled with high genetic advance indicated their governance by additive gene action. A negative significant correlation was observed between protein content and grain yield. Mahalanobis D 2 analysis grouped the 36 genotypes into eight clusters. Clustering pattern failed to indicate any relationship between genetic diversity and geographic diversity. Based on genetic diversity and performance, the genotypes MS 1168, MS 174 and CO 13 were found to be suitable for use as parents in a hybridization program for improving yield; the genotypes MS 1168, MS 174 and MS 2869 for protein and Malawi 1915 and CO 11 for calcium. Protein and calcium contents contributed less to genetic divergence.     

Variation in mineral micronutrient concentrations in grain of wheat lines of diverse origin

150 lines of bread wheat representing diverse origin and 25 lines of durum, spelt, einkorn and emmer wheat species were analysed for variation in micronutrient concentrations in grain. A subset of 26 bread wheat lines was grown at six sites or seasons to identify genetically determined differences in micronutrient concentrations. Substantial variation among the 175 lines existed in grain Fe, Zn and Se concentrations. Spelt, einkorn and emmer wheats appeared to contain higher Se concentration in grain than bread and durum wheats. Significant differences between bread wheat genotypes were found for grain Fe and Zn, but not Se concentration; the latter was influenced more by the soil supply. Grain Zn, but not Fe, concentration correlated negatively with grain yield, and there was a significant decreasing trend in grain Zn concentration with the date of variety release, suggesting that genetic improvement in yield has resulted in a dilution of Zn concentration in grain. Both grain Zn and Fe concentrations also correlated positively and significantly with grain protein content and P concentration, but the correlations with kernel size, kernel weight or bran yield were weak. The results from this study are useful for developing micronutrient biofortification strategies.     

Correlation of Plant Morphological and Grain Quality Traits with Mineral Element Contents in Yunnan Rice

Correlations between four grain quality, 24 plant morphological traits and eight element contents of 653 accessions from Yunnan rice were analyzed. P, K, Ca and Mg contents of core collection were closely correlated to the most plant morphological and grain quality traits, and there were highly significant correlations （P 〈0.01） among some traits including P content to number of stems and tillers, K content and amylose content, Ca content and plant height, Mg content and protein content. Mn, Zn, Cu and Fe contents of core collection were closely related to a few traits, such as Fe content and gel consistency （-0.1121^＊＊）, Zn content and seed setting rate （-0.1411^＊＊）, Cu content and number of grains per panicle （-0.1398^＊＊）, Mn content and plant height （-0.2492^＊＊）.     

北京地区不同年代冬小麦品种籽粒微量元素含量的变化及其对水分的响应

为探讨北京地区小麦品种更替过程中籽粒微量元素含量的演变规律,选用20世纪60年代以来小麦品种更替过程中的7个主要推广品种,分别种植在春不浇水和春浇二水条件下,测定了小麦籽粒中Fe、Zn、Mn、Cu、蛋白质含量以及籽粒产量.结果表明,北京地区小麦品种更替过程中,随着产量的提高,籽粒微量元素含量并没有出现明显的稀释效应,而且有些营养成分含量(如Fe、Mn)随着年代更替出现明显提高的趋势;春不浇水降低了籽粒Fe、Zn、Cu和蛋白质含量,但提高了籽粒Mn含量.相关分析表明,小麦籽粒Fe、Zn、Cu含量与蛋白质含量呈正相关,Fe、Zn、Mn、Cu、蛋白质含量与籽粒产量、穗粒数和千粒重以及年代之间也呈正相关,说明在产量改良的过程中有望同步提高籽粒微量元素和蛋白质含量.     

Influence of long-term nitrogen fertilization on micronutrient density in grain of winter wheat (Triticum aestivum L.)

A long-term (1999–2007) field experiment was conducted to investigate the effects of three nitrogen (N) fertilization rates (0, 130, and 300 kg N/ha) on micronutrient density in wheat grain and its milling fractions. At maturity, grains were harvested and fractionated into flour, shorts, and bran for micronutrient and N analysis. N fertilization increased iron (Fe), zinc (Zn), and copper (Cu) density in wheat grain compared to the control. Increase of N application rate from 130 to 300 kg N/ha, however, did not further increase the three micronutrient densities in grain. Micronutrient concentrations were usually highest in the bran and lowest in the flour. High N application increased Zn and Cu densities in all three milling fractions and increased Fe concentration in shorts and bran but not in flour. N application did not affect the manganese (Mn) concentration in grain. N fertilization changed the proportions of Fe and Cu in flour and bran but did not affect the distribution of Zn. Because N fertilization increased micronutrient accumulation in wheat grain, proper management of N fertilization has the potential to enhance the nutritional quality of this important food.     

Effects of Different Nitrogen Fertilizer Levels and Native Soil Properties on Rice Grain Fe, Zn and Protein Contents

Deposition of protein and metal ions (Fe, Zn) in rice grains is a complex polygenic trait showing considerable environmental effect. To analyze the effect of nitrogen application levels and native soil properties on rice grain protein, iron (Fe) and zinc (Zn) contents, 32 rice genotypes were grown at three different locations each under 80 and 120 kg/hm2 nitrogen fertilizer applications. In treatments with nitrogen fertilizer application, the brown rice grain protein content (GPC) increased significantly (1.1% to 7.0%) under higher nitrogen fertilizer application (120 kg/hm2) whereas grain Fe/Zn contents showed non-significant effect of nitrogen application level, thus suggesting that the rate of uptake and translocation of macro-elements does not influence the uptake and translocation of micro-elements. The pH, organic matter content and inherent Fe/Zn levels of native soil showed significant effects on grain Fe and Zn contents of all the rice genotypes. Grain Zn content of almost all the tested rice genotypes was found to increase at Location III having loamy soil texture, neutral pH value (pH 6.83) and higher organic matter content than the other two locations (Locations I and II), indicating significant influence of native soil properties on brown rice grain Zn content while grain Fe content showed significant genotype × environment interaction effect. Genotypic difference was found to be the most significant factor to affect grain Fe/Zn contents in all the tested rice genotypes, indicating that although native soil properties influence phyto-availability of micronutrients and consequently influencing absorption, translocation and grain deposition of Fe/Zn ions, yet genetic makeup of a plant determines its response to varied soil conditions and other external factors. Two indica rice genotypes R-RF-31 (27.62 μg/g grain Zn content and 7.80% GPC) and R1033-968-2-1 (30.05 μg/g grain Zn content and 8.47% GPC) were identified as high grain Zn and moderate GPC rice genotypes. These results indicate that soil property and organic matter content increase the availability of Fe and Zn in rhizosphere, which in turn enhances the uptake, translocation and redistribution of Fe/Zn into rice grains.     

氮、磷、锌营养对水稻籽粒植酸含量的影响及与几种矿质元素间的相关性

以协青早、秀水110及其辐射诱变获得的低植酸突变系（HIPi1和HIPj1）为材料,通过水培试验对不同氮、磷、锌浓度处理下水稻籽粒植酸含量差异及与几种矿质元素间的相关性进行了比较分析。高水平氮、磷、锌浓度处理的籽粒植酸含量较同一品种的低氮、磷、锌处理均有所降低,但在水稻生育期间,籽粒植酸含量对磷、锌处理浓度变化的敏感性,则因品种的植酸类型特征而异;氮、磷浓度增加能分别提高铁或降低铜在籽粒中的积累,但在高锌处理下,籽粒铁含量明显降低、而钾和镁的含量等却有所升高; 籽粒植酸含量一般与K、Mg、Fe、Cu 4种矿质元素含量呈正相关、与籽粒Zn含量呈负相关,但统计显著水平因品种而异。低植酸突变体籽粒中的K、Mg、Fe、Zn等含量虽略有下降,但可以通过适当的介质营养条件来调节有关矿质营养在水稻籽粒中的积累。     

QTL Analysis for Grain Iron and Zinc Concentrations in Two O. nivara Derived Backcross Populations

Identification of quantitative trait loci (QTLs) for grain mineral elements can assist in faster and more precise development of micronutrient dense rice varieties through marker-assisted breeding. In the present study, QTLs were mapped for Fe and Zn concentrations in two BC₂F₃ mapping populations derived from the crosses of O. sativa cv Swarna with two different accessions of O. nivara. In all, 10 and 8 QTLs were identified for grain Fe and Zn concentrations in population 1, and 7 and 5 QTLs were identified in population 2, respectively. Eighty percent of the QTLs detected in both populations were derived from O. nivara. Five QTLs for Fe and three QTLs for Zn explained more than 15% phenotypic variance either in interval or composite interval mapping. The locations of O. nivara derived QTLs such as qFe2.1, qFe3.1, qFe8.2 and qZn12.1 were consistently identified in both the populations. Epistatic interaction was observed only between RM106 and RM6 on chromosome 2 and between RM22 and RM7 on chromosome 3 for Fe concentration in population 1. Sixteen candidate genes for metal homeostasis were found to co-locate with 10 QTLs for Fe and Zn concentrations in both the populations. Most of the Fe and Zn QTLs were found to co-locate with QTLs for grain yield and grain quality traits. Some of the major effect QTLs identified can be used to improve rice grain Fe and Zn concentrations.     

Generation mean analysis of pearl millet [Pennisetum glaucum (L.) R. Br.] grain iron and zinc contents and agronomic traits in West Africa

Pearl millet is the most important staple food crop for millions of people across the world. Micronutrient malnutrition is the major problem for people living in the semi-arid regions of Africa. Identification of gene effects controlling the inheritance of grain Fe and Zn will be helpful in formulating suitable breeding strategies for biofortified pearl millet development. Hence, generation mean analysis was used to study epistasis and estimate gene effects for grain iron and zinc contents along with the agronomic and morphological traits. Six generations P₁, P₂, F₁, F₂, BC₁P₁ and BC₁P₂ were generated and were evaluated during the 2018–19 off season. Analysis of variance showed significant variability for all the traits in both generations. Six parameter model revealed predominance of additive gene effects for inheritance of grain iron concentration, and additive × additive type of non-allelic interactions. For grain zinc concentration additive gene effects were preponderant compared to non-additive gene effects, and only additive × dominance gene effects were significant among the three types of epistasis. Grain weight per plant was predominantly under non-additive gene effects and additive × additive and additive × dominance gene effects type of epistasis was detected in each cross. Likewise, for flowering non-additive gene effects were most important with the presence of dominance × dominance type of epistasis. For plant height, panicle circumference and length, additive × additive genes effects were the most important among the three type of non-allelic gene action.These findings can be helpful in enehancing the pearl millet breeding programs in Africa.     

Grain concentrations of protein and mineral nutrients in a large collection of spelt wheat grown under different environments

A large number of spelt wheat genotypes (ranging from 373 to 772) were evaluated for grain concentrations of protein and mineral nutrients under 6 different environments. There was a substantial genotypic variation for the concentration of mineral nutrients in grain and also for the total amount of nutrients per grain (e.g., content). Zinc (Zn) showed the largest genotypic variation both in concentration (ranging from 19 to 145 mg kg⁻¹) and content (ranging from 0.4 to 4.1 μg per grain). The environment effect was the most important source of variation for grain protein concentration (GPC) and for many mineral nutrients, explaining between 37 and 69% of the total sums of squares. Genotype by environment (G × E) interaction accounted for between 17 and 58% of the total variation across the minerals. GPC and sulfur correlated very significantly with iron (Fe) and Zn. Various spelt genotypes have been identified containing very high grain concentrations of Zn (up to 70 mg kg⁻¹), Fe (up to 60 mg kg⁻¹) and protein (up to 30%) and showing high stability across various environments. The results indicated that spelt is a highly promising source of genetic diversity for grain protein and mineral nutrients, particularly for Zn and Fe.     

Agronomic characteristics, grain quality and flour rheology of 372 bread wheats in a worldwide core collection

A core collection of 372 accessions representative of worldwide hexaploid bread wheat diversity [Balfourier, F., Roussel, V., Strelchenko, P., Exbrayat-Vinson, F., Sourdille, P., Boutet, G., Koenig, J., Ravel, C., Mitrofanova, O., Beckert, M., Charmet, G., 2007. A worldwide bread wheat core collection arrayed in a 384-well plate. Theoretical and Applied Genetics 114, 1265–1275] was used to evaluate the available genetic diversity of agronomic and quality characteristics. The traits assessed during the vegetative period were date of ear-emergence, date of flowering, lodging, disease susceptibility and pre-harvest sprouting. Thousand kernel weight, test weight, grain hardness, grain protein content, pentosan viscosity and grain colour were also measured. The rheological properties of the derived white flours were estimated using mixograph and alveograph tests. For most of the traits, a wide phenotypic variation was observed across all the accessions. Several parameters (mixograph width parameters before and after peak time, alveograph dough tenacity and extensibility, near infrared measurements, like those for protein content, and absorbance measurements of palmitic acid and linoleic acid content) made it easier to discriminate between the cultivars. The largest ranges of variation were found in landraces and old cultivars rather than in more recent varieties. This is evidence that there is sufficient variability available for rare alleles, which have been eliminated in breeding modern varieties to be detected. Such a core collection will therefore be a useful resource for future genetic studies on wheat quality.     

Agronomic characteristics,grain quality and flour rheology of 372 bread wheats in a worldwide core collection

A core collection of 372 accessions representative of worldwide hexaploid bread wheat diversity [Balfourier, F., Roussel, V., Strelchenko, P., Exbrayat-Vinson, F., Sourdille, P., Boutet, G., Koenig, J., Ravel, C., Mitrofanova, O., Beckert, M., Charmet, G., 2007. A worldwide bread wheat core collection arrayed in a 384-well plate. Theoretical and Applied Genetics 114, 1265–1275] was used to evaluate the available genetic diversity of agronomic and quality characteristics. The traits assessed during the vegetative period were date of ear-emergence, date of flowering, lodging, disease susceptibility and pre-harvest sprouting. Thousand kernel weight, test weight, grain hardness, grain protein content, pentosan viscosity and grain colour were also measured. The rheological properties of the derived white flours were estimated using mixograph and alveograph tests. For most of the traits, a wide phenotypic variation was observed across all the accessions. Several parameters (mixograph width parameters before and after peak time, alveograph dough tenacity and extensibility, near infrared measurements, like those for protein content, and absorbance measurements of palmitic acid and linoleic acid content) made it easier to discriminate between the cultivars. The largest ranges of variation were found in landraces and old cultivars rather than in more recent varieties. This is evidence that there is sufficient variability available for rare alleles, which have been eliminated in breeding modern varieties to be detected. Such a core collection will therefore be a useful resource for future genetic studies on wheat quality.     

Genome wide association studies (GWAS) of element contents in grain with a special focus on zinc and iron in a world collection of barley (Hordeum vulgare L.)

Genome wide association studies (GWAS) were carried out to map Quantitative Trait Loci (QTL) associated with element contents in the grain using 336 spring barley. Of the elements analyzed, Fe content ranged from 21.9 to 91.0 mg kg⁻¹, Zn from 10.4 to 54.5 mg kg⁻¹, Ba from 0.2 to 8.9, Ca from 186.4 to 977.5, Cu from 1.5 to 9.8, K from 353.2 to 7721.5, Mg from 1049.8 to 2024.2, Mn from 8.1 to 22.9, Na from 55.9 to 627.9, P from 2272.9 to 5428.8, S from 880.7 to 1898.0, Si from 19.1 to 663.2, and Sr from 0.35 to 2.62 mg kg⁻¹. GWAS were carried out using 6519 SNP markers and multiple elements in MLM:PCA + K model in TASSEL software. Population analyses showed two sub-populations, primarily based on row types. GWAS for row types showed association with INTERMEDIUM-C, a modifier gene for lateral spikelet fertility in the 4H chromosome, validating current GWAS approach. GWAS also showed that 2 QTL for Ba, 2 for Ca, 4 for Cu, 11 for Fe, 2 for K, 3 for Mg, 6 for Mn, 4 for Na, 3 for S, 5 for Si, and 3 for Zn were mapped in barley chromosomes. The QTL identified in the current study are valuable for breeding nutrient dense barley cultivars in the future, especially Zn and Fe.     

Grain concentrations of protein and mineral nutrients in a large collection of spelt wheat grown under different environments

A large number of spelt wheat genotypes (ranging from 373 to 772) were evaluated for grain concentrations of protein and mineral nutrients under 6 different environments. There was a substantial genotypic variation for the concentration of mineral nutrients in grain and also for the total amount of nutrients per grain (e.g., content). Zinc (Zn) showed the largest genotypic variation both in concentration (ranging from 19 to 145 mg kg⁻¹) and content (ranging from 0.4 to 4.1 μg per grain). The environment effect was the most important source of variation for grain protein concentration (GPC) and for many mineral nutrients, explaining between 37 and 69% of the total sums of squares. Genotype by environment (G × E) interaction accounted for between 17 and 58% of the total variation across the minerals. GPC and sulfur correlated very significantly with iron (Fe) and Zn. Various spelt genotypes have been identified containing very high grain concentrations of Zn (up to 70 mg kg⁻¹), Fe (up to 60 mg kg⁻¹) and protein (up to 30%) and showing high stability across various environments. The results indicated that spelt is a highly promising source of genetic diversity for grain protein and mineral nutrients, particularly for Zn and Fe.