Classification of rice genotypes based on their mechanisms of adaptation to iron toxicity

Iron (Fe) toxicity is a nutritional disorder that affects lowland rice (Oryza sativa L.). The occurrence of excessive amounts of reduced Fe(II) in the soil solution, its uptake by the rice roots, and its transpiration‐driven transport result in elevated Fe(II) concentrations in leaf cells that catalyze the formation of reactive oxygen species. The oxidative stress causes rusty brown spots on leaves (bronzing) and the reduction of biomass and yield. While the use of resistant genotypes is the most promising approach to address the problem, the stress appears to differentially affect rice plants as a function of plant age, climatic conditions, stress intensity and duration, and the prevailing adaptation mechanism. We comparatively assessed 21 contrasting 6‐week‐old rice genotypes regarding their response (symptom score, biomass, Fe concentrations and uptake) to a 6 d iron pulse of 1500 mg L^–1 Fe(II). Eight selected genotypes were further compared at different stress intensities (0, 500, 1000, and 1500 mg L^–1 Fe(II)) and at different developmental stages (4‐, 6‐, and 8‐week‐old plants). Based on Fe‐induced biomass reduction and leaf‐bronzing score, the tested spectrum was grouped in resistant and sensitive genotypes. Linking bronzing scores to leaf iron concentrations allowed further differentiation into includer and excluder types. Iron precipitation on roots and organ‐specific iron partitioning permitted to classify the adaptation strategies into root exclusion, stem and leaf sheath retention, and leaf blade tissue tolerance. The effectiveness of these strategies differed with stress intensity and developmental stage. The reported findings improve the understanding of Fe‐stress response and provide a basis for future genotype selection or breeding for enhancing Fe‐toxicity resistance in rice.     

Analysis of Genotype,Environment, and Their Interaction Effects on the Phytochemicals and Antioxidant Capacities of Red Rice (Oryza sativa L.)

Fourteen red rice varieties were planted in two locations during summer (Hangzhou) and winter (Hainan) to study the effect of genotype and environment on the phytochemicals and antioxidant capacities of rice grain. B‐type proanthocyanidins in red rice were detected by LC‐MS/MS and quantified by using the vanillin assay. Analysis of variance showed that total phenolic content (TPC), total flavonoid content (TFC) and 2,2′‐azino‐bis‐(3‐ethylbenzothiazoline‐6‐sulfonic acid) (ABTS) radical scavenging capacity were mainly affected by environmental factors, which accounted for more than 60% of the total variance. However, total proanthocyanidin content (TPAC) and 1,1‐diphenyl‐2‐picrylhydrazyl (DPPH) radical scavenging capacity were equally affected by both genotype and environment. The genotype × environment effects were significant for all traits. The pairwise correlations among TPC, TFC, TPAC, ABTS, and DPPH were also significant (r > 0.900, P < 0.001). Principal component analysis identified the genotypes that had higher contents of antioxidants and more stability across environments. This study showed that indirect selection of a simple trait (i.e., TPC) is an effective way to select rice high in antioxidant capacity in breeding programs. This study also suggests that rice should be produced specifically in a certain environment for the end user to minimize the variation in the functional properties and maximize their contents.     

Comparison of iron bioavailability from 15 rice genotypes: studies using an in vitro digestion/caco-2 cell culture model

An in vitro digestion/Caco-2 model was used to compare iron bioavailability from 15 selected Fe-dense and normal genotypes of unpolished rice from the International Rice Research Institute. Iron uptake was determined using Caco-2 cell ferritin formation in response to exposure to a digest of the cooked rice. Iron bioavailabilities from all rice genotypes were ranked as a percent relative to a control variety (Nishiki). Iron concentration in the rice samples ranged from 14 to 39 microg/g. No correlation was observed between Fe uptake and grain-Fe concentration. Furthermore, phytic acid levels were not correlated with Fe bioavailability. Genotypes with low Fe bioavailability (Tong Lan Mo Mi, Zuchein, Heibao, and Xua Bue Nuo) were noticeably more brown to purple in color. The results suggest that certain unknown compounds related to rice grain color may be a major factor limiting Fe bioavailability from unpolished rice.     

Genotypic differences in concentrations of iron,manganese, copper,and zinc in polished rice grains

Identification of genotypic differences in micronutrient concentrations of staple food crops is essential if plant breeding strategies are to improve human mineral nutrition. The concentrations of zinc (Zn), iron (Fe), copper (Cu), and manganese (Mn) in polished grains of 285 rice (Oryza sativa L.) genotypes and the relationship between concentrations of the four micronutrient elements and concentrations of protein and lysine were examined. Significant differences (P<.01) were found in the concentrations of Zn, Fe, Cu, and Mn in polished rice with a fairly normal distribution among rice genotypes. On average, Cu and Zn concentrations of Indica rice were about 2‐fold higher than Japonica rice, while Fe concentrations of Japonica rice were slightly higher than Indica rice. Among Indica rice genotypes, red rice contained higher Zn than white rice. Protein and lysine concentrations differed considerably among the genotypes, but no close relationship between the micronutrients and protein or lysine concentrations was observed among genotypes. Sixteen genotypes with significantly higher grain Zn, Fe, Cu, and Mn concentrations were identified.     

A quick and efficient screen for resistance to iron toxicity in lowland rice

Iron (Fe) toxicity is a major stress to rice in many lowland environments worldwide. Due to excessive uptake of Fe²⁺ by the roots and its acropetal translocation into the leaves, toxic oxygen radicals may form and damage cell structural components, thus impairing physiological processes. The typical visual symptom is the “bronzing” of the rice leaves, leading to substantial yield losses, particularly when toxicity occurs during early vegetative growth stages. The problem is best addressed through genotype improvement, i.e., tolerant cultivars. However, the time of occurrence and the severity of symptoms and yield responses vary widely among soil types, years, seasons, and genotypes. Cultivars resistant in one system may fail when transferred to another. Better targeting of varietal improvement requires selection tools improving our understanding of the resistance mechanisms and strategies of rice in the presence of excess iron. A phytotron study was conducted to develop a screen for seedling resistance to Fe toxicity based on individual plants subjected to varying levels of Fe (0–3000 mg L^–1 Fe supplied as Fe(II)SO₄), stress duration (1–5 d of exposure), vapor‐pressure deficit (VPD; 1.1 and 1.8 kPa), and seedling age (14 and 28 d). Genotypes were evaluated based on leaf‐bronzing score and tissue Fe concentrations. A clear segregation of the genotypic tolerance spectrum was obtained when scoring 28 d old seedlings after 3 d of exposure to 2000 mg L^–1 Fe in a high‐VPD environment. In most cases, leaf‐bronzing scores were highly correlated with tissue Fe concentration (visual differentiation in includer and excluder types). The combination of these two parameters also identified genotypes tolerating high levels of Fe in the tissue while showing only few leaf symptoms (tolerant includers). The screen allows selecting genotypes with low leaf‐bronzing score as resistant to Fe toxicity, and additional analyses of the tissue Fe concentration of those can identify the general adaptation strategy to be utilized in breeding programs.     

Physiological investigations of management and genotype options for adapting rice production to iron toxicity in Madagascar

Iron (Fe) toxicity is one of the major mineral disorders affecting rice (Oryza sativa L.) production in Madagascar. This study aimed at linking physiological and agronomic responses of diverse rice genotypes to Fe resistance mechanisms with different nutrient management practices. Twenty‐three local and exotic rice varieties were grown in Fe‐toxic soil in parallel greenhouse and field experiments and subjected to two treatments: (1) no fertilizer; (2) mineral and organic fertilizer application at recommended rates. Growth, straw and grain yield, symptom formation, and physiological responses including Fe uptake, root plaque formation, and lipid peroxidation were monitored. The application of fertilizer significantly decreased average shoot Fe concentrations partly due to Fe exclusion favored by enhanced root plaque formation. Visual symptoms negatively correlated with straw biomass in both experiments and grain yield in the greenhouse experiment, and positively correlated with lipid peroxidation. However, no plausible correlation occurred with grain yield in the field due to sterility in exotic varieties un‐adapted to local climate. Even though grain Fe concentrations were orders of magnitude lower than in vegetative tissue, some exotic varieties were significantly superior to local checks. Our results provide insight into management and genotype options for adapting rice to Fe toxicity under field conditions.     

Diversity in Phytochemical Composition and Antioxidant Capacity of Dent,Flint, and Specialty Corns

Dent, flint, and specialty genotypes of Indian yellow maize were evaluated for phytochemical content and their hydrophilic antioxidant capacity. Solid‐phase extraction coupled with solid–liquid extraction was used to obtain free phenolics and flavonoids from maize samples, reducing the use of excessive solvents and handling time. Bound phenolics were extracted with enhanced acidic hydrolysis to improve extractability. The phenolic contents in Indian maize genotypes ranged from 46 to 79 μmol of ferulic acid equivalents per gram of sample on a dry basis (db). Carotenoids in Indian maize genotypes were found to be equivalent to Chinese yellow maize and ranged from 13 to 24 μg/g of sample (db), whereas tocol content varied greatly in the range of 607–1,238 μg of α‐tocopherol equivalents/g of sample (db). Dent and flint corn did not exhibit differences (P > 0.05) in their phenolic contents, whereas among the specialty genotypes sweet corn contained the highest phenolics but least carotenoids (P < 0.05). Bound extracts appeared to contribute largely to 2,2′‐diphenyl‐1‐picrylhydrazyl free radical scavenging, hydrogen peroxide scavenging, and ferric reducing antioxidant power of maize genotypes. Sweet corn exhibited higher antioxidant capacity (P < 0.05) among all the genotypes. The antioxidant capacities correlated well with the total phenolic contents of the maize genotypes.     

Farm-scale variations in soil color as influenced by organic matter and iron oxides in Japanese paddy fields

To understand how soil color is influenced by soil components at the farm scale, we evaluated spatial variation in soil color and related soil properties in Japanese paddy fields. After harvest of rice, 246 surface soil samples were collected in 10-m grids from five contiguous irrigated paddy fields, each with an area of about 0.5 ha. The samples were analyzed to determine color parameters (L*, a*, and b*), and contents of total C, total N, Fe oxides, sand, and loss-on-ignition. The results obtained were modeled and mapped geostatistically. All color parameters indicated strong spatial dependence with long ranges (>85 m). In contrast, total C and N showed short ranges (about 40 m). The contents of Fe oxides, sand, and loss-on-ignition showed intermediate ranges (50–85 m). The ranges of these properties and their distribution patterns suggested that the contents of total C and N were influenced by long-term application of manure and that sand content was influenced by topography and past land consolidation. Further soil color analysis after removal of organic matter or silt plus clay particles revealed that soil organic matter, texture, and Fe oxides affected soil color parameters in a complex manner. Prediction of total C from soil darkness was hindered by the presence of silt plus clay particles containing Fe oxides. On the other hand, citrate-dithionite extractable Fe was estimated accurately from the b* value (yellowness), which can be useful for predicting the occurrence of akiochi (autumn decline) disease of rice at the farm scale.     

An automatic procedure for determination of available iron in Indian soils

Abstract

An on‐line automatic procedure for the flow injection analysis (FIA) determination of iron (Fe) in a variety of soil extracts with common laboratory reagents, i.e., thiocyanate, thiocyanate +1,10‐phenanthroline/bipyridine has been tested. The apparent molar absorptivity of the complexes lies in the range of (5.5–7.0)x10³ L mole^‐1 cm^‐1 at an absorption maximum between 470–495 nm. The detection limit of the method is 15 ppb Fe. The sample output is 100 samples/hr. Almost all ions associated with Fe in the soil extract do not interfere in this method. Optimization of FIA variables, composition of the complex, and effect of other ions on the determination of Fe are discussed. The method has been used for determination of Fe status in a variety of agricultural soils in east Madhya Pradesh, India.     

Iron deficiency stress response of various c‐3 and c‐4 grain crop genotypes: Strategy II mechanism evaluated

The relative amount of phytosiderophore produced by various Strategy II plants has been categorized as follows: barley (Hordeum vulgare L.) > wheat (Triticum aestivum L.) > oat (Avena byzantina C. Koch.) > rye (Secale cereale L.) >> corn (Zea mays L.) >> sorghum (Sorghum bicolor (L.) Moench) > rice (Oryza sativa L.). With the exception of rice, these plants developed under oxidized soil conditions, and the C‐3 species produce more phytosiderophore than C‐4 species under Fe‐deficiency stress. Iron‐efficient Coker 227 oat produced phytosiderophore in response to Fe‐deficiency stress, while Fe‐inefficient TAM 0–312 oat did not. Although Fe‐efficient WF9 corn and Fe‐inefficient ys₁ corn differed in their ability to obtain Fe, neither produced sufficient quantities of phytosiderophore to explain these differences. The objectives of this research were to determine: (a) if phytosiderophore production of Fe‐deficiency stressed C‐4 species millet (Panicum miliaceum L.) and corn is low or absent compared to identically stressed C‐3 species oat and barley, and (b) if native, inbred and hybrid corn cultivars differ in ability to produce and utilize phytosiderophores.

Although release of phytosiderophore for Fe‐stressed corn and millet was generally lower than oat, quantity of release was not always related to obtaining Fe and maintaining green plants. Barley maintained high leaf Fe and low chlorosis with a minor release of phytosiderophore. Oat with increased release acted similarly to barley, whereas a relatively high release of phytosiderophore from White maize did not effect Fe uptake or greening. Likewise, small amounts of phytosiderophore were produced by all corn types, but corn was generally unable to obtain adequate Fe from the growth medium. Two of the native corns, Coneso and Tepecintle, maintained relatively low chlorosis, but they differed in phytosiderophore release. Thus, it appears that the C‐4 plants studied herein generally release a lower amount of phytosiderophore than do C‐3 species, but overcoming Fe‐deficiency chlorosis is not guaranteed by such release. The Strategy II mechanism of mere release of phytosiderophore and consequential Fe acquisition appears simplistic. There is a need for understanding what other factors are involved.     

Interaction of Oxidation Products from Caffeic Acid with Fe(III) and Fe(II)

Abstract

Phenolic substances in the soil–plant system can be oxidized by metal ions, inorganic components, molecular oxygen as well as by phenoloxidases, giving rise to the formation of products of low or high molecular weight. Interactions of these products with iron, in both reduced and oxidized form, can affect the iron mobility in soil and rhizosphere, and thus its availability to plants. Here we report the results of a study on the complexing and reducing activity of the oxidation products from caffeic acid (CAF), obtained via electrochemical means, towards Fe(III) and Fe(II) in aqueous solution in the 3.0–6.0 pH range. The HPLC analysis of the filtered solutions after the CAF oxidation showed the formation of two main groups of products: (i) CAF oligomers formed through radicalic reactions which do not involve the double bond of the CAF lateral chain and (ii) products where this bond is involved. These oxidation products (COP) were found to interact with both Fe(III) and Fe(II) with formation of soluble and insoluble Fe(III)‐, and Fe(II)‐COP complexes. The COP were found to be able to reduce Fe(III) to Fe(II) mainly at pH < 4.0. A low redox activity was observed at pH ≥ 4.5 due to Fe(III) hydrolysis reactions as well as to the decrease in the redox potential of the Fe(III)/Fe(II) couple. Formation of hydroxy Fe(III)‐COP polymers occurs at pH > 3.5.     

Physiological characteristics of selenite uptake by maize roots in response to different pH levels

In selenite solutions, H₂SeO₃, HSeO , and SeO<$>_3^{2‐}<$> are in equilibrium in proportions that vary with solution pH. The physiological characteristics of selenite uptake were studied with excised roots of maize (Zea mays L.) seedlings at pH 3.0, 5.0, and 8.0. The results showed that 0.10 mM 2,4‐dinitrophenol (DNP), 1.0 mM sodium fluoride (NaF), and a temperature of 4°C inhibited selenite uptake by maize roots by 16%, 20%, and 23% at pH 3.0, by up to 80%, 79%, and 78% at pH 5.0, and by 5%, 9%, and 16% at pH 8.0. Hence, selenite may enter roots at pH 5.0 in an energy‐dependent manner, in contrast to pH 3.0 and 8.0. The uptake kinetics for selenite were determined for excised roots of maize, and the curves were linear at pH 3.0 and 8.0, but saturated at pH 5.0, showing that carrier‐mediated uptake of selenite occurred at pH 5.0, but not at pH 3.0 or 8.0. Further studies showed that HgCl₂ and AgNO₃ inhibited selenite uptake separately by 81% and 76% at pH 3.0 and indicated that selenite was absorbed by maize roots through aquaporins at pH 3.0. At pH 8.0, anion‐channel inhibitors only inhibited a small fraction of selenite uptake, indicating that the major absorption pathway of SeO<$>_3^{2‐}<$> species into roots was not absorbed passively through anion channels, but might involve other processes. According to these results, it is proposed that selenite uptake occurs via different mechanisms depending on its species in solution in response to pH levels.     

Magnesium deficiency–induced impairment of photosynthesis in leaves of fruiting Citrus reticulata trees accompanied by up‐regulation of antioxidant metabolism to avoid photo‐oxidative damage

Limited data are available on the physiological responses of leaves from fruiting trees to magnesium (Mg) deficiency. Magnesium deficiency–induced effects on photosystem II (PSII) photochemistry in leaves of fruiting (Citrus reticulate cv. Ponkan) trees were assessed by the chlorophyll a fluorescence (OJIP) transient. Magnesium deficiency decreased leaf CO₂ assimilation and carbohydrates, but had no effect on intercellular CO₂ concentration. Activity of ribulose‐1,5‐bisphosphate carboxylase/oxygenase (Rubisco) and concentrations of Chlorophyll (Chl) and carotenoids (Car) decreased to a lesser extent than CO₂ assimilation. Chlorophyll a fluorescence transient from Mg‐deficient leaves had increased O step and decreased P step, accompanied by positive ΔL, ΔK, ΔJ, and ΔI bands. Magnesium deficiency decreased maximum quantum yield of primary photochemistry (F_v/F_m), quantum yield of electron transport from Q<$>_A^‐<$> to the photosystem I (PSI) end electron acceptors (φ_R0), maximum amplitude of IP phase and total performance index (PI_{tot, abs}), but increased deactiviation of oxygen‐evolving complex (OEC) and energy dissipation. Magnesium‐deficient leaves had higher or similar activities of antioxidant enzymes except for lower catalase (CAT) activity, higher or similar concentrations of antioxidant metabolites, and a higher ratio of Car : Chl. Magnesium‐deficiency did not affect concentration of malondialdehyde (MDA) and ratios of ascorbate (ASC) to ASC + dehydroascorbate (DHA) and reduced glutathione (GSH) to GSH + oxidized glutathione (GSSG). In conclusion, Mg deficiency–induced impairment of the whole photosynthetic electron transport chain may be the main factor contributing to decreased CO₂ assimilation. Enhanced energy dissipation and antioxidant metabolism provide sufficient protection to Mg‐deficient leaves against photo‐oxidative damage.     

Interactive effects of salinity and iron deficiency on different rice genotypes

Salinity adversely affects plant growth, photosynthesis, and availability of nutrients including iron. Rice (Oryza sativa L.) is susceptible to soil salinity and highly prone to iron (Fe) deficiency due to lower release of Fe‐chelating compounds under saline conditions. In order to investigate the effects of salinity and low iron supply on growth, photosynthesis, and ionic composition of five rice genotypes (KS‐282, Basmati Pak, Shaheen Basmati, KSK‐434 and 99417), a solution culture experiment was conducted with four treatments (control, 50 mM NaCl, Fe‐deficient, and 50 mM NaCl + Fe‐deficient). Salinity and Fe deficiency reduced shoot and root growth, photosynthetic and transpiration rates, chlorophyll concentration, and stomatal conductance. The reduction in all these parameters was more in the interactive treatment of salinity and low Fe supply. Moreover, a significant increase in shoot and root Na⁺ with corresponding decrease in K⁺ and Fe concentrations was also observed in the combined salinity and Fe‐deficiency treatment. Among the tested genotypes, Basmati Pak was the most sensitive genotype both under salt stress and Fe deficiency. The genotype KS‐282 performed better than other genotypes under salinity stress alone, whereas Shaheen Basmati was the best genotype under Fe deficiency in terms of all the studied parameters.     

A Rapid Microassay for Determination of Peroxidase in Wheat and Flour

A rapid sensitive microplate assay for the determination of peroxidase activity in wheat, flour, or individual wheat kernels has been developed using a commercially prepared 2,2′‐azinobis(3‐ethylbenzothiazoline‐6‐sulfonic acid) diammonium salt (ABTS) solution. The assay derives a six‐point calibration curve based on commercially available horseradish peroxidase (r² > 0.990), while simultaneously analyzing eight extracts in triplicate. The coefficients of variation (CV) of triplicate assays of a single extract from any of the three sources investigated were generally <2.0%. Multiple extracts (n = 5) of either whole meal or flour yielded assays with an average CV < 5.0%. Preparation of the calibration standards with commercially available peroxidase stabilizing buffer allowed the standards to be used for five days without any deterioration in the assay's reproducibility. This assay is ideally suited for high‐throughput operations such as millstream analysis or plant breeder screening evaluations.     

Phytosiderophore release as a criterion for genotypic evaluation of iron efficiency in oat

Iron (Fe) deficiency in small grains grown on calcareous soils results in reduced yields, is difficult and expensive to treat with fertilizer, and is complicated to overcome by genetic field screening due to heterogeneous soil and environmental conditions. Recently, phytosiderophore release has been linked to ability of species and genotypes to resist Fe‐deficiency chlorosis. We propose a laboratory technique to measure phytosiderophore release by Fe‐deficient oat (Avena sativa L.) genotypes as a selection method for Fe‐deficiency chlorosis resistance in oat. Plants were grown in Fe‐limiting nutrient solution and phytosiderophore release was measured on 11 days. Summations of daily phytosiderophore release by 17 oat genotypes correlate well with Fe‐deficiency chlorosis scores in the field (r = ‐0.70, p = 0.01). The proposed method consistently identified the genotypes most susceptible to Fe deficiency but did not clearly separate the moderately susceptible genotypes. In these latter genotypes, other factors such as active uptake sites, root growth rate, utilization of acquired Fe, or soil interactions may be modifying factors to phytosiderophore in Fe efficiency. Quantification of phytosiderophore provides a useful selection criterion for oat by eliminating the most inefficient types and with refinement, may become a powerful tool for identifying Fe efficiency in grass crops.     

Atrazine photodegradation in aqueous solution induced by interaction of humic acids and iron: photoformation of iron(II) and hydrogen peroxide

The photochemical formation of Fe(II) and hydrogen peroxide (H 2O 2) coupled with humic acids (HA) was studied to understand the significance of iron cycling in the photodegradation of atrazine under simulated sunlight. The presence of HA significantly enhanced the formation of Fe(II) and H 2O 2, and their subsequent product, hydroxyl radical ( (*)OH), was the main oxidant responsible for the atrazine photodegradation. During 60 h of irradiation, the fraction of iron presented as Fe(II) (Fe(II)/Fe(t)) decreased from 20-32% in the presence of the Fe(III)-HA complex to 10-22% after adding atrazine. The rate of atrazine photodegradation in solutions containing Fe(III) increased with increasing HA concentration, suggesting that the complexation of Fe(III) with HA accelerated the Fe(III)/Fe(II) cycling. Using fluorescence spectrometry, the quenching constant and the percentage of fluorophores participating in the complexation of HA with Fe(III) were estimated by the modified Stern-Volmer equation. Fourier transform infrared spectroscopy (FTIR) offered the direct evidence that Fe(III)-carboxylate complex could be formed by ligand exchange of HA with Fe(III). Based on all the information, a possible reaction mechanism was proposed.     

Physiological basis of iron chlorosis tolerance in rice (Oryza sativa) in relation to the root exudation capacity

Micronutrient deficiency in cultivable soil, particularly that of iron (Fe) and zinc (Zn), is a major productivity constraint in the world. Low Fe availability due to the low solubility of the oxidized ferric forms is a challenge. An experiment was, thus, executed to assess the performance of eight genetically diverse rice genotypes on Fe-sufficient (100 µM) and Fe-deficient (1 µM) nutrient solution, and their ability to recover from Fe deficiency was measured. Fe efficiency under Fe deficiency in terms of biomass production showed a significant positive correlation with the root release of phytosiderophore (PS) (R² = 0.62*). This study shows that the Fe deficiency tolerance of Pusa 33 was related to both a high release of PS by the root and an efficient translocation of Fe from the root to the shoot as the Fe–PS complex, which could be useful for improving the Fe nutrition of rice particularly under aerobic conditions.     

Responses of aerobically grown iron chlorosis tolerant and susceptible rice (Oryza sativa L.) genotypes to soil iron management in an Inceptisol

Iron deficiency is a serious nutritional disorder in aerobic rice, causing chlorosis, poor yields and reduced grain nutritional quality. The problem can be managed by complementing the use of Fe-efficient plant type with a suitable Fe management strategy. In the present paper, we report the effect of eight iron management practices to resolve the problem of iron (Fe) chlorosis through the use of an iron deficiency tolerant (IDTR) and iron deficiency susceptible (IDSR) rice genotype, i.e. Pusa 33 and ADT 39, respectively. Fe deficiency tolerance of these genotypes was related to the root release of PS which enabled a higher uptake of Fe in the IDTR than the IDSR under Fe deficiency. In general, IDTR performed better than the IDSR as evident from a significant increase in total iron, active iron, chlorophyll content and grain and straw yield. IDSR produced the highest grain and straw yield under slow iron release nano clay complex source. Grain Fe content of the IDTR and IDSR increased by 18.9 and 13.4%, respectively, under recommended dose of Fe. The results identified the most effective soil management strategies for the alleviating Fe deficiency chlorosis and improving Fe nutrition of both IDTR and IDSR genotypes.     

Distribution of Protein Bodies and Phytate‐Rich Inclusions in Grain Tissues of Low and High Iron Rice Genotypes

The present study aims to understand whether genotypic differences in grain iron (Fe) concentration in four rice genotypes are related to its association with protein bodies containing phytate‐rich inclusions. Rice genotypes with high and low grain Fe concentrations in unpolished brown rice were grown in a greenhouse at Chiang Mai, Thailand, and grains were harvested at maturity. The presence of protein bodies and phytate‐rich inclusions in rice grain tissues were examined by means of light and transmission electron microscopy (TEM). The composition of mineral elements in different grain tissues was examined using energy dispersive X‐ray microanalysis (EDX) and chemical analysis. The relative distribution pattern of protein bodies in the tissues was similar among the four rice genotypes, which resembled the pattern of grain N concentrations in these tissues. The high grain Fe genotypes (based on brown rice Fe concentration) had more protein bodies containing phytate‐rich inclusions in the embryo and aleurone layer tissues than the low Fe genotypes. Phytate‐rich inclusions were not detected in the endosperm tissues in all genotypes. In conclusion, the presence of protein bodies with phytate‐rich inclusions predominantly in the embryo and aleurone regions of the grain is an important parameter contributing to the variation in brown rice Fe concentration among the genotypes, but not in the white rice (the endosperm). Iron associated with the phytate‐rich inclusions present in the embryo and aleurone layer tissues are largely lost during the polishing process to produce white rice.