Nicotianamine synthase gene expression differs in barley and rice under Fe-deficient conditions

Nicotianamine (NA) is an intermediate in the biosynthetic pathway of the mugineic acid family phytosiderophores (MAs), which are crucial components of the iron acquisition apparatus of graminaceous plants. In non-graminaceous plants, NA is thought to be an essential chelator for metal cation homeostasis. Thus NA plays a key role in Fe metabolism and homeostasis in all higher plants. Nicotianamine synthase (NAS, EC 2.5.1.43) catalyzes the trimerization of S-adenosylmethionine to form one molecule of NA. Barley, a plant that is resistant to Fe deficiency, secretes large amounts of MAs, whereas rice, a plant that is susceptible to Fe deficiency, secretes only small amounts. In this study we isolated a genomic fragment containing HvNAS1 from barley and three rice cDNA clones, osnas1, osnas2 and osnas3, from Fe-deficient rice roots. We also isolated a genomic fragment containing both OsNAS1 and OsNAS2. In contrast to barley, in which Fe deficiency induces the expression of NAS genes only in roots, Fe deficiency in rice induced NAS gene expression in both roots and chlorotic leaves. The amounts of endogenous NA in both the roots and leaves were higher than in barley. We introduced barley genomic DNA fragments containing HvNAS1 with either 9 or 2 kb of the 5'-flanking region into rice, using Agrobacterium-mediated transformation. Fe deficiency induced HvNAS1 expression in both roots and leaves of the transgenic rice, as occurs with rice NAS genes. Barley and rice NAS genes are compared in a discussion of alteration of the NAS genes during adaptation to Fe deficiency.     

Aspects of the Fe-and Mn nutrition of rice plants

Summary The combination of low Mn levels and high Fe levels in tissues of lowland rice varieties, as often encountered when rice is grown on acid soils, is not likely to result from an antagonistic effect of Fe on the uptake of Mn.Experiments with rice plants growing on sand, supplied with Fe and Mn, and subjected to various pH levels and moisture regimes, made it clear that under acid anaerobic conditions the absorption of Mn by rice plants is little affected by the presence of large quantities of Fe, and that under acid aerobic conditions the absorption of Fe by rice plants is little affected by the presence of large quantities of Mn.     

Iron,zinc and manganese nutrition of wetland rice (Oryza sativa L.) on a gypsum amended sodic soil

A field experiment was conducted to evaluate the effect of three levels of Fe and two levels of Zn, and their combinations, on the growth, yield and Fe, Zn, and Mn nutrition of rice on a zinc deficient sodic soil amended with gypsum. Iron and zinc were supplied as sulphates. Application of Zn significantly enhanced the yield of rice and available soil and plant Zn irrespective of Fe application. Maximum response of rice to Zn was obtained when Fe was applied at the highest rate. While Fe application brought about a significant improvement in available soil and plant Fe and Mn, it decreased significantly Zn content of the crop. After crop harvest, recovery of added Fe was 20% and Zn 12%. Results suggest that benefits of Fe application to rice in sodic soils can only be realised if it is applied along with Zn.     

Understanding Fe2+ toxicity and P deficiency tolerance in rice for enhancing productivity under acidic soils

Plants experience low phosphorus (P) and high iron (Fe) levels in acidic lowland soils that lead to reduced crop productivity. A better understanding of the relationship between these two stresses at molecular and physiological level will lead to development of suitable strategies to increase crop productivity in such poor soils. Tolerance for most abiotic stresses including P deficiency and Fe toxicity is a quantitative trait in rice. Recent studies in the areas of physiology, genetics, and overall metabolic pathways in response to P deficiency of rice plants have improved our understanding of low P tolerance. Phosphorous uptake and P use efficiency are the two key traits for improving P deficiency tolerance. In the case of Fe toxicity tolerance, QTLs have been reported but the identity and role played by underlying genes is just emerging. Details pertaining to Fe deficiency tolerance in rice are well worked out including genes involved in Fe sensing and uptake. But, how rice copes with Fe toxicity is not clearly understood. This review focuses on the progress made in understanding these key environmental stresses. Finally, an opinion on the key genes which can be targeted for this stress is provided.     

有色稻米Fe、Zn、Cu和Mn含量及与种皮颜色相关分析

用AAS方法测定了弥勒县相同生态条件下种植的27份有色稻和34份普通稻糙米4种矿质元素含量,并对有色米和普通米Fe、Zn、Cu和Mn含量进行了比较研究。结果表明,有色稻米4种矿质元素含量明显高于无色稻米,其差异均达显著水平,其含量高低依次为Zn>Fe>Cu>Mn;对黑、褐、红、黄、绿5种不同种皮颜色的稻米4种矿质元素含量进行比较研究,发现稻米Fe含量(mg/kg)依次为黑>绿>褐>红>黄,Zn含量(mg/kg)依次为绿>红>黑>褐>黄,Cu含量(mg/kg)依次为黑>褐>红>黄>绿,Mn含量(mg/kg)依次为褐>黑>红>黄>绿;并且Fe和Mn含量在不同颜色稻米间差异均达显著水平,与有色米种皮颜色密切相关,而Zn和Cu差异不显著,与有色米种皮颜色关系不大。黑米和褐米富Fe、Zn、Cu和Mn,绿米富Fe和Zn,红米富Zn和Cu,黄米4种矿质元素含量较低,Fe、Cu和Mn均低于普通稻米。     

Rice plants take up iron as an Fe3+-phytosiderophore and as Fe2+

Only graminaceous monocots possess the Strategy II iron (Fe)-uptake system in which Fe is absorbed by roots as an Fe3+-phytosiderophore. In spite of being a Strategy II plant, however, rice (Oryza sativa) contains the previously identified Fe2+ transporter OsIRT1. In this study, we isolated the OsIRT2 gene from rice, which is highly homologous to OsIRT1. Real-time PCR analysis revealed that OsIRT1 and OsIRT2 are expressed predominantly in roots, and these transporters are induced by low-Fe conditions. When expressed in yeast (Saccharomyces cerevisiae) cells, OsIRT2 cDNA reversed the growth defects of a yeast Fe-uptake mutant. This was similar to the effect of OsIRT1 cDNA. OsIRT1- and OsIRT2-green fluorescent protein fusion proteins localized to the plasma membrane when transiently expressed in onion (Allium cepa L.) epidermal cells. OsIRT1 promoter-GUS analysis revealed that OsIRT1 is expressed in the epidermis and exodermis of the elongating zone and in the inner layer of the cortex of the mature zone of Fe-deficient roots. OsIRT1 expression was also detected in the ccompanion cells. Analysis using the positron-emitting tracer imaging system showed that rice plants are able to take up both an Fe3+-phytosiderophore and Fe2+. This result indicates that, in addition to absorbing an Fe3+-phytosiderophore, rice possesses a novel Fe-uptake system that directly absorbs the Fe2+, a strategy that is advantageous for growth in submerged conditions.     

Nicotianamine,a Novel Enhancer of Rice Iron Bioavailability to Humans

Background

Polished rice is a staple food for over 50% of the world''s population, but contains little bioavailable iron (Fe) to meet human needs. Thus, biofortifying the rice grain with novel promoters or enhancers of Fe utilization would be one of the most effective strategies to prevent the high prevalence of Fe deficiency and iron deficiency anemia in the developing world.

Methodology/Principal Findings

We transformed an elite rice line cultivated in Southern China with the rice nicotianamine synthase gene (OsNAS1) fused to a rice glutelin promoter. Endosperm overexpression of OsNAS1 resulted in a significant increase in nicotianamine (NA) concentrations in both unpolished and polished grain. Bioavailability of Fe from the high NA grain, as measured by ferritin synthesis in an in vitro Caco-2 cell model that simulates the human digestive system, was twice as much as that of the control line. When added at 1∶1 molar ratio to ferrous Fe in the cell system, NA was twice as effective when compared to ascorbic acid (one of the most potent known enhancers of Fe bioavailability) in promoting more ferritin synthesis.

Conclusions

Our data demonstrated that NA is a novel and effective promoter of iron utilization. Biofortifying polished rice with this compound has great potential in combating global human iron deficiency in people dependent on rice for their sustenance.     

Phytoavailability of phosphate adsorbed on ferrihydrite,hematite, and goethite

Poorly crystalline Fe in soil has been shown to affect Fe and P availability. Oxalate extractable Fe, a measure of poorly crystalline Fe oxides, has not been compared to soil test methods for Fe and P in rice soils. Twenty eight soils used for rice production were incubated under aerobic and anaerobic soil conditions and extracted for Fe and P with ammonium oxalate, ammonium acetate-EDTA (AA-EDTA), ammonium bicarbonate-DTPA (AB-DTPA) and DTPA. Citrate-dithionite extractable Fe and Fe content of rice plants in a greenhouse experiment were also determined. Soils used in this experiment had a large amount of poorly-crystalline Fe oxide. In some soils, poorly-crystalline Fe constituted 60% of the citrate-dithionite extractable Fe. The amount of extractable Fe and P increased significantly under anaerobic conditions. The relationships between extractants showed that DTPA Fe was highly correlated to AB-DTPA Fe and oxalate Fe was highly correlated to AA-EDTA Fe. There was no relationship between Fe and P extracted by AB-DTPA, while there was a better relationship with ammonium oxalate and AA-EDTA extractants. Poorly-crystalline Fe and P extracted by ammonium oxalate were correlated.     

Mutation in nicotianamine aminotransferase stimulated the Fe(II) acquisition system and led to iron accumulation in rice

Higher plants acquire iron (Fe) from the rhizosphere through two strategies. Strategy II, employed by graminaceous plants, involves secretion of phytosiderophores (e.g. deoxymugineic acid in rice [Oryza sativa]) by roots to solubilize Fe(III) in soil. In addition to taking up Fe in the form of Fe(III)-phytosiderophore, rice also possesses the strategy I-like system that may absorb Fe(II) directly. Through mutant screening, we isolated a rice mutant that could not grow with Fe(III)-citrate as the sole Fe source, but was able to grow when Fe(II)-EDTA was supplied. Surprisingly, the mutant accumulated more Fe and other divalent metals in roots and shoots than the wild type when both were supplied with EDTA-Fe(II) or grown under water-logged field conditions. Furthermore, the mutant had a significantly higher concentration of Fe in both unpolished and polished grains than the wild type. Using the map-based cloning method, we identified a point mutation in a gene encoding nicotianamine aminotransferase (NAAT1), which was responsible for the mutant phenotype. Because of the loss of function of NAAT1, the mutant failed to produce deoxymugineic acid and could not absorb Fe(III) efficiently. In contrast, nicotianamine, the substrate for NAAT1, accumulated markedly in roots and shoots of the mutant. Microarray analysis showed that the expression of a number of the genes involved in Fe(II) acquisition was greatly stimulated in the naat1 mutant. Our results demonstrate that disruption of deoxymugineic acid biosynthesis can stimulate Fe(II) acquisition and increase iron accumulation in rice.