Overexpression of the Barley Nicotianamine Synthase Gene HvNAS1 Increases Iron and Zinc Concentrations in Rice Grains

In humans, iron (Fe) and zinc (Zn) deficiencies result in major worldwide health problems. Transgenic technologies to produce Fe- and Zn-biofortified rice varieties offer a promising potential solution. Nicotianamine, the precursor of phytosiderophores, chelates Fe²⁺ and Zn²⁺ and plays an important role in transporting these metals to both vegetative and reproductive organs within the plant. Our objective was to increase Fe and Zn contents in rice grains by overexpressing the barley nicotianamine synthase gene HvNAS1. HvNAS1-overexpressing transgenic rice showed increased HvNAS1 expression and subsequent increases in endogenous nicotianamine and phytosiderophore content in shoots, roots, and seeds. Fe and Zn concentrations in polished T₁ seeds from transgenic plants increased more than three and twofold, respectively; Fe and Zn concentrations also increased in both polished and brown T₂ seeds. These results suggest that the overproduction of nicotianamine enhances the translocation of Fe and Zn into rice grains.     

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.     

Agronomic fortification of rice grains with secondary and micronutrients under differing crop management and soil moisture regimes in the north Indian Plains

Although the System of Rice Intensification (SRI) has been reported to produce higher paddy (Oryza sativa L.) yields with better-quality grains, little research has addressed the latter claim. This study investigated the interactive effects of rice cultivation methods with different irrigation schedules and plant density on the uptake and concentration of sulfur (S), zinc (Zn), iron (Fe), manganese (Mn) and copper (Cu) in the grain and straw of two rice cultivars during two rainy seasons in the northern plains of India. As the two seasons differed in amounts of rainfall, there were impacts of soil moisture differences on nutrient uptake. Plots with SRI cultivation methods enhanced the grain uptake and concentrations of S, Zn, Fe, Mn and Cu by 36, 32, 28, 32 and 63%, respectively, compared to conventional transplanting (CT). Under SRI management, the highest concentrations of S, Zn and Cu in the grain and straw occurred with irrigation intervals scheduled at 3 days after disappearance of ponded water (DADPW; 3_D), whereas Fe and Mn concentrations in the grain and straw were higher with irrigation at 1 DADPW (1 _D ) compared with plots under 3 _D or 5 DADPW (5 _D ). The higher nutrient uptakes were also manifested in higher grain yield in 1 _D and 3 _D plots (by 9 and 6%, respectively) compared with 5 _D . Wider spacing (25 × 25 cm) compared with closer spacing (20 × 20 cm) significantly increased yield and the uptake and concentrations of all the said nutrients in the grains. When comparing the performance of two cultivars, the total uptakes of Zn, Fe, Mn and Cu in both grain and straw were significantly more in Hybrid 6444 than the improved variety Pant Dhan 4. Overall, SRI crop management compared to CT practices led to more biological fortification of rice grains with respect to S and the four micronutrients studied, giving a concomitant yield advantage of about 17% on average in this region.     

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。     

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.     

不同锌效率基因型水稻籽粒中矿质元素的原位微区分布研究

 采用同步辐射（SRXRF）结合ICP MS研究了足Zn条件下不同锌效率水稻籽粒中Zn及其他矿质元素的微区分布特征。结果表明SRXRF技术可高效分析水稻籽粒锌等矿质元素的微区分布特征。水稻籽粒中K和Ca元素的分布特征与其余4种元素相关不显著,而Zn、Fe、Cu、Mn元素的含量分布相互间则显著相关;水稻籽粒中K含量最高,移动性也最强,精米中K含量可达颖壳的52.6%~90.6%,而Ca在颖壳中含量最高。其余矿质元素含量最高值均出现在糊粉层;Zn、Fe等元素主要集中在水稻籽粒的胚及糊粉层等外层功能组织,在胚乳中由外向里呈递减趋势。此外,锌高效基因型IR8192在水稻籽粒各部位的Zn含量均低于锌低效基因型二九丰。可见,与锌低效品种相比,锌高效品种仅在低Zn水平下表现出更高的锌吸能力及利用效率,在足Zn条件下并无籽粒富锌特征。     

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.     

Protein,energy and mineral utilization in rats fed rice-legume diets

Protein, energy and mineral utilization were examined in growing rats fed cooked, dry legume seeds in combination with polished rice. The legumes tested included 3 varieties of common beans, black, white and brown (Phaseolus vulgaris), lentils (Lens esculenta) and peas (Pisum sativum). The rice:legume mixtures (1:1 N ratio) were the only dietary sources of protein, Zn, Fe and Cu. The rice:black bean mixture was also tested at a 4:1 N ratio. Nutrient utilization was studied by balance trials and mineral utilization was further assessed by tissue analyses. True protein digestibility (TD) and energy digestibility of the rice:legume (1:1) mixtures were high. Biological value (BV) varied only little but was lowest in the rice:lentil mixture. The rice:brown bean mixture had the lowest amount of utilizable protein (UP). The (1:1) rice:legume mixtures did not adversely affect Fe status, as measured by liver Fe content and blood hemoglobin and hematocrit, or Cu status, as measured by liver Cu content and plasma ceruloplasmin. However, femur zinc content indicated a sub-optimal Zn status. Increasing the dietary N from rice in combination with black beans resulted in a substantial improvement of BV, NPU and Zn status of the animals but had a negative effect on UP and Fe status of the rats.     

Variations in Concentration and Distribution of Health-Related Elements Affected by Environmental and Genotypic Differences in Rice Grains

Malnutrition is one of the major problems inmost of the developing countries, especially amongwomen, infants and children. Biofortification is a verynew strategy to enhance the bioavailability ofmicronutrients in staple food by using advancedbreeding meth…     

Losses of essential mineral nutrients by polishing of rice differ among genotypes due to contrasting grain hardness and mineral distribution

The effect of different polishing techniques on loss of mineral elements from rice grains was quantified using a panel of indica and tropical japonica genotypes, previously classified as differing in ease of polishing. Gradients in mineral elements across the bran-endosperm interface were quantified using micro-scaled precision abrasive polishing in combination with inductively coupled plasma mass spectrometry and synchrotron X-ray fluorescence microscopy. Frictional polishing, similar to that of commercial mills, i.e. 8–10% loss of grain weight, reduced the concentration of Fe, Mg, P, K and Mn by 60–80% in all genotypes. Following gentler polishing (3–5% weight loss), genotypes classified as difficult to polish showed smaller decreases in Fe, Mg, P, K and Mn compared to genotypes classified as easy to polish. The concentration of other elements, e.g. Zn, S, Ca, Cu, Mo and Cd, showed comparable reductions (<30%) irrespective of polishing technique or ease of polishing. The different patterns of polishing losses of minerals reflected their distribution within the grain. Five-fold differences in the reduction of Zn concentration during polishing were observed for different genotypes which started with similar Zn concentrations in the unpolished grain, thus showing clear potential for selecting genotypes with reduced polishing losses of Zn.     

Barley Bulgur: Effect of Processing and Cooking on Chemical Composition

Three barley cultivars (Hordeum vulgareL.) were processed into bulgur by pressure cooking or cooking at atmospheric pressure. The effect of processing on levels of thiamine, riboflavin, minerals (Fe, Cu, Zn, Mn, Ca, Mg) as well as the phytic acid and β-glucan was investigated. Significant decreases (p<0·05) were observed in ash, riboflavin and thiamine contents during bulgur processing. Neither the cooking methods nor the dehulling process had significant influence on the content of Fe, Cu, Zn or Mg. However, the Mn and Ca content of the bulgurs were significantly (p<0·05) lower compared with the corresponding raw barleys. For all cultivars total P and phytate P contents of the bulgurs were significantly (p<0·05) lower compared with the corresponding raw barleys. In contrast, levels of β-glucan were significantly higher in processed bulgur vs raw barley. Protein contents of the samples did not change significantly during bulgur processing. Bulgur processed from barley appeared to retain most of the nutritional value of raw barley, in particular it showed high levels of soluble dietary fibre.     

Determinants of barley grain yield in a wide range of Mediterranean environments

Barley grain yield in rainfed Mediterranean regions can be largely influenced by terminal drought events. In this study the ecophysiological performance of the ‘Nure’ (winter) × ‘Tremois’ (spring) barley mapping population (118 Doubled Haploids, DHs) was evaluated in a multi-environment trial of eighteen site–year combinations across the Mediterranean Basin during two consecutive harvest years (2004 and 2005). Mean grain yield of sites ranged from 0.07 to 5.43 t ha⁻¹, clearly dependent upon both the total water input (rainfall plus irrigation) and the water stress index (WSI) accumulated during the growing season. All DHs were characterized for possessing molecular marker alleles tagging four genes that regulate barley cycle, i.e. Vrn-H1, Vrn-H2, Ppd-H2 and Eam6. Grain yield differences were initially interpreted in terms of mean differences between genotypes (G), environments (E), and for each combination of genotype and environment (GE) through a “full interaction” ANOVA model. Variance components estimates clearly showed the greater importance of GE over G, although both were much lower than E. Alternative linear and bilinear models of increasing complexity were used to describe GE. A linear model fitting allelic variation at the four genes explained genotype main effect and genotype × environment interaction much better than growth habit itself. Adaptation was primarily driven by the allelic constitution at three out of the four segregating major genes, i.e. Vrn-H1, Ppd-H2 and Eam6. In fact, the three genes together explained 47.2% of G and 26.3% of GE sum of squares. Grain yield performance was more determined by the number of grains per unit area than by the grain weight (phenotypic correlation across all genotypic values: r = 0.948 and 0.559, respectively). The inter-relationships among a series of characters defining grain yield and its components were also explored as a function of the length of the different barley developmental phases, i.e. vegetative, reproductive, and grain filling stages. In most environments, the best performing (adapted) genotypes were those with faster development until early occurrence of anthesis. This confirmed the crucial role of the period defining the number of grains per unit area in grain yield determination under Mediterranean environments.     

Phytate and mineral elements concentration in a collection of Italian durum wheat cultivars

Mineral deficiencies are prevalent in human populations and the improvement of the mineral content in cereal products represents a possible strategy to increase the human mineral intake. Nevertheless, most of the inorganic phosphorus (P_i) present in mature cereal seeds (40–80%) is stored as phytate, an anti-nutritional factor that forms complexes with minerals such as Ca, Mg, Zn and Fe reducing their bioavailability. The present study was undertaken: (i) to determine the variation in phytate and mineral concentrations in the whole grains of 84 Italian durum wheat (Triticum durum Desf.) cultivars representative of old and modern germplasm; (ii) to estimate the magnitude of genotype × environment interaction effects; and (iii) to examine the interrelationships among mineral concentrations in durum wheat with the final aim to identify superior durum wheat cultivars that possess low phytate content and high concentration of mineral elements in their whole-wheat flour. The cultivars were grown in field trials during 2004–2005 at Foggia, Italy and during 2005–2006 at Foggia and Fiorenzuola d’Arda—Southern and Northern Italy. The phytate content was estimated indirectly by using a microtitre plate assay evaluating the P_i absorbance at 820 nm, while the Cu, Fe, Mn, Ca, K, Mg, Na and Zn mineral contents were determined by ICP/OES. The contents of Zn and Fe across years and locations ranged from 28.5 to 46.3 mg/kg for Zn with an average of 37.4 mg/kg and from 33.6 to 65.6 mg/kg for Fe with an average of 49.6 mg/kg. P_i grain content was between 0.46 and 0.76 mg/g showing a positive correlation with all minerals except Cu and Zn. Although breeding activity for Fe and Zn would be difficult because G × E interaction is prevalent, multi-location evaluation of germplasm collection help to identify superior genotypes to achieve this objective. The results here reported open the possibility of designing a specific breeding program for improving the nutritional value of durum wheat through the identification of parental lines with low-P_i and high minerals concentration in whole grains.     

A Wheat Xylanase Inhibitor Protein (XIP-I) Accumulates in the Grain and has Homologues in Other Cereals

We have measured xylanase inhibitor activity against an Aspergillus niger xylanase in different parts of the wheat plant at different stages of development and used immunodetection to determine the spatial and temporal distribution of xylanase inhibitor protein I (XIP-I) in Triticum aestivum var. Soisson. Although xylanase inhibitor activity was detected in all parts of the wheat plant throughout development, XIP-I was located predominantly in the grain tissue, where it appears at a late stage in grain development and persists after germination, indicating that the different xylanase inhibitor proteins are under different regulatory controls. (1,4)-β-Xylanase activity was detected in wheat grains during development and post-germination. Pure XIP-I and a crude sample containing TAXI inhibitors but not XIP-I did not have the ability to inhibit this endogenous (1,4)-β-xylanase activity. Protein extracts from the seeds of various monocots were also tested for the presence of XIP-I, where it was found to be present in the grains of several wheat varieties, rye and barley, but was not detected in rice, sorghum or maize.