Using iron fertilizer to control Cd accumulation in rice plants: A new promising technology

Effects of two kinds of iron fertilizer, FeSO4 and EDTA·Na2Fe were studied on cadmium accumulation in rice plants with two rice genotypes, Zhongzao 22 and Zhongjiazao 02, with soil culture systems. The results showed that application of iron fertilizers could hardly make adverse effects on plant growth and rice grain yield. Soil application of EDTA·Na2Fe significantly reduced the Cd accumulation in rice roots, shoots and rice grain. Cd concentration in white rice of both rice genotypes in the treatment of soil application of EDTA·Na2Fe was much lower than 0.2 mg/kg, the maximal Cd permission concentra- tion in cereal crop foods in State standard. However, soil application of FeSO4 or foliar application of FeSO4 or EDTA·Na2Fe resulted in the significant increase of Cd accumulation in rice plants including rice grain compared with the control. The results also showed iron fertilizers increased the concentra- tion of iron, copper and manganese element in rice grain and also affected zinc concentration in plants. It may be a new promising way to regulate Cd accumulation in rice grain in rice production through soil application of EDTA·Na2Fe fertilizers to maintain higher content of available iron and ferrous iron in soils.     

Using iron fertilizer to control Cd accumulation in rice plants: A new promising technology

Effects of two kinds of iron fertilizer, FeSO₄ and EDTA·Na₂Fe were studied on cadmium accumulation in rice plants with two rice genotypes, Zhongzao 22 and Zhongjiazao 02, with soil culture systems. The results showed that application of iron fertilizers could hardly make adverse effects on plant growth and rice grain yield. Soil application of EDTA·Na₂Fe significantly reduced the Cd accumulation in rice roots, shoots and rice grain. Cd concentration in white rice of both rice genotypes in the treatment of soil application of EDTA·Na₂Fe was much lower than 0.2 mg/kg, the maximal Cd permission concentration in cereal crop foods in State standard. However, soil application of FeSO₄ or foliar application of FeSO₄ or EDTA·Na₂Fe resulted in the significant increase of Cd accumulation in rice plants including rice grain compared with the control. The results also showed iron fertilizers increased the concentration of iron, copper and manganese element in rice grain and also affected zinc concentration in plants. It may be a new promising way to regulate Cd accumulation in rice grain in rice production through soil application of EDTA·Na₂Fe fertilizers to maintain higher content of available iron and ferrous iron in soils.     

水稻胚乳过表达VIT1/VIT2对Fe和Cd积累的影响

水稻籽粒铁(Fe)缺乏和镉(Cd)含量超标是农业生产亟待解决的重要问题。以往研究表明,OsVIT1和OsVIT2是液泡铁转运蛋白,本研究选取野生型ZH11为背景材料,使用胚乳特异性表达启动子Glb-1构建了胚乳过表达OsVIT1和OsVIT2材料。RT-qPCR分析表明,OsVIT1在转化植株的胚乳和叶片过量表达,OsVIT2在转化植株的胚乳过量表达。通过田间试验,研究胚乳过表达OsVIT1和OsVIT2对水稻不同部位Fe和Cd积累的影响。结果表明,胚乳过表达OsVIT1显著降低籽粒中的Fe浓度约50%,显著增加秸秆的锌(Zn)、铜(Cu)浓度和籽粒中的Cu浓度,胚乳过表达OsVIT2显著降低籽粒中的Fe、Cd浓度约50%,显著增加秸秆的Fe浓度45%–120%。胚乳过表达OsVIT1和OsVIT2不影响水稻的农艺性状。总之,胚乳过表达OsVIT1和OsVIT2降低了水稻籽粒的Fe积累,未达到预期效果,胚乳过表达OsVIT2还降低籽粒的Cd积累,增加秸秆Fe积累,为水稻铁生物强化和降镉提供了借鉴。     

Root iron plaque formation and characteristics under N2 flushing and its effects on translocation of Zn and Cd in paddy rice seedlings (Oryza sativa)

Background and Aims

Anoxic conditions are seldom considered in root iron plaque induction of wetland plants in hydroponic experiments, but such conditions are essential for root iron plaque formation in the field. Although ferrous ion availability and root radial oxygen loss capacity are generally taken into account, neglect of anoxic conditions in root iron plaque formation might lead to an under- or over-estimate of their functional effects, such as blocking toxic metal uptake. This study hypothesized that anoxic conditions would influence root iron plaque formation characteristics and translocation of Zn and Cd by rice seedlings.

Methods

A hydroponic culture was used to grow rice seedlings and a non-disruptive approach for blocking air exchange between the atmosphere and the induction solution matrix was applied for root iron plaque formation, namely flushing the headspace of the induction solution with N₂ during root iron plaque induction. Zn and Cd were spiked into the solution after root iron plaque formation, and translocation of both metals was determined.

Key Results

Blocking air exchange between the atmosphere and the nutrient solution by N₂ flushing increased root plaque Fe content by between 11 and 77 % (average 31 %). The N₂ flushing treatment generated root iron plaques with a smoother surface than the non-N₂ flushing treatment, as observed by scanning electron microscopy, but Fe oxyhydroxides coating the rice seedling roots were amorphous. The root iron plaques sequestrated Zn and Cd and the N₂ flushing enhanced this effect by approx. 17 % for Zn and 71 % for Cd, calculated by both single and combined additions of Zn and Cd.

Conclusions

Blocking of oxygen intrusion into the nutrient solution via N₂ flushing enhanced root iron plaque formation and increased Cd and Zn sequestration in the iron plaques of rice seedlings. This study suggests that hydroponic studies that do not consider redox potential in the induction matrices might lead to an under-estimate of metal sequestration by root iron plaques of wetland plants.     

Silicon induces phytochelatin and ROS scavengers facilitating cadmium detoxification in rice

• Cadmium (Cd) is detrimental to crops and the environment. This work examines the natural mechanisms underlying silicon‐ (Si‐)directed Cd detoxification in rice plants.
• The addition of Si to plants under Cd stress caused significant improvements in morphological parameters, chlorophyll score, F_v/F_m and total soluble protein concentration compared to controls, confirming that Si is able to ameliorate Cd‐induced damage in rice plants. This morpho‐physiological evidence was correlated with decreased cell death and electrolyte leakage after Si application.
• The results showed no critical changes in root Cd concentration, while shoot Cd decreased significantly after Si supplementation in comparison with Cd‐stressed rice. Additionally, expression of Cd transporters (OsNRAMP5 and OsHMA2) was significantly down‐regulated while the concentration of phytochelatin, cysteine and glutathione, together with expression of OsPCS1 (phytochelatin synthase) in roots of Cd‐stressed rice was significantly induced when subjected to Si treatment. This confirms that the alleviation of Cd stress is not only limited to the down‐regulation of Cd transporters but also closely related to the phytochelatin‐driven vacuolar storage of Cd in rice roots.
• The enzymatic analysis further revealed the role of SOD and GR enzymes in protecting rice plants from Cd‐induced oxidative harm. These findings suggest a mechanistic basis in rice plants for Si‐mediated mitigation of Cd stress.

     

Role of iron plaque in Cd uptake by and translocation within rice (Oryza sativa L.) seedlings grown in solution culture

A hydroponics culture experiment was conducted to investigate the effect of iron plaque on Cd uptake by and translocation within rice seedlings grown under controlled growth chamber conditions. Rice seedlings were pre-cultivated for 43 days and then transferred to nutrient solution containing six levels of Fe (0, 10, 30, 50, 80 and 100 mg L⁻¹) for 6 days to induce different amounts of iron plaque on the root surfaces. Seedlings were then exposed to solution containing three levels of Cd (0, 0.1 and 1.0 mg L⁻¹) for 4 days. In order to differentiate the uptake capability of Cd by roots with or without iron plaque, root tips (white root part without iron plaque) and middle root parts (with iron plaque) of pre-cultivated seedlings treated with 0, 30 and 50 mg L⁻¹ Fe were exposed to ¹⁰⁹Cd for 24 h. Reddish iron plaque gradually became visible on the surface of rice roots but the visual symptoms of the iron plaque on the roots differed among treatments. In general, the reddish color of the iron plaque became darker with increasing Fe supply, and the iron plaque was more homogeneously distributed all along the roots. The Fe concentrations increased significantly with increasing Fe supply regardless of Cd additions. The Cd concentrations in dithionite–citrate–bicarbonate (DCB)-extracts and in shoots and roots were significantly affected by Cd and Fe supply in the nutrient solution. The Cd concentrations increased significantly with increasing Cd supply in the solution and were undetectable when no Cd was added. The Cd concentrations in DCB-extracts with Fe supplied tended to be higher than that at Fe₀ at Cd_0.1, and at Cd_1.0, DCB-Cd with Fe supplied was significantly lower. Cd concentrations in roots and shoots decreased with increasing Fe supply at both Cd additions. The proportion of Cd in DCB-extracts was significantly lower than in roots or shoots. Compared to the control seedlings without Fe supply, the radioactivity of ¹⁰⁹Cd in shoots of seedlings treated with Fe decreased when root tips were exposed to ¹⁰⁹Cd and did not change significantly when middle parts of roots were exposed. Our results suggest that root tissue rather than iron plaque on the root surface is a barrier to Cd uptake and translocation within rice plants, and the uptake and translocation of Cd appear to be related to Fe nutritional levels in the plants.     

Iron plaque formation and its effect on arsenic uptake by different genotypes of paddy rice

Background and aims

Iron plaque on roots has been hypothesized to be an effective restraint on the uptake of arsenic (As) by rice plants. Evaluating the formation of iron plaque and its effect on As uptake by various rice cultivars is valuable because selecting low As uptake rice cultivars results in reduced risks associated with rice consumption. This study examines iron plaque formation and its effect on As uptake by different genotypes of rice cultivars.

Methods

Hydroponic cultures were conducted in phytotron at day 25/night 20°C and the rice seedlings in fifth-leaf age were treated with Fe (II) at the levels of 0 and 100 mg L^?1 in the Kimura B nutrient solutions for 14 days. The amount of iron plaque formation of 28 rice cultivars was determined by using the DCB extractable Fe of roots. Four cultivars representing high and low iron plaque formation capability, from indica and japonica respectively, were selected out of the 28 cultivars and processed for Fe and As treatments. After Fe treatments for 4 days, the seedlings were fed with As (III) at levels of 0, 0.5, and 1 mg L^?1 for another 10 days. We were thus able to determine the amounts of iron plaque formation and the As content in iron plaque, roots, and shoots of the four tested cultivars.

Results

Iron plaque formation capability differed among tested twenty-eight rice cultivars. Feeding As to four tested cultivars enhanced iron plaque formation on roots; the As uptake by roots and shoots was decreased by the addition of Fe. Both the retention of As on iron plaque and the decrease of As uptake by the addition of Fe varied among tested cultivars and were not correlated with the iron plaque formation capability.

Conclusions

Iron plaque can sequestrate As on the roots and reduce rice’s As uptake. However, other factors also influence the As uptake, namely the differences in binding affinity of iron plaque to As, the existent As species in the rhizosphere, and the uptake capability of various As species by rice plants. These factors should also be considered when selecting low As uptake rice cultivars.     

The iron-chelate transporter OsYSL9 plays a role in iron distribution in developing rice grains

Key message

Rice OsYSL9 is a novel transporter for Fe(II)-nicotianamine and Fe(III)-deoxymugineic acid that is responsible for internal iron transport, especially from endosperm to embryo in developing seeds.

Abstract

Metal chelators are essential for safe and efficient metal translocation in plants. Graminaceous plants utilize specific ferric iron chelators, mugineic acid family phytosiderophores, to take up sparingly soluble iron from the soil. Yellow Stripe 1-Like (YSL) family transporters are responsible for transport of metal-phytosiderophores and structurally similar metal-nicotianamine complexes. Among the rice YSL family members (OsYSL) whose functions have not yet been clarified, OsYSL9 belongs to an uncharacterized subgroup containing highly conserved homologs in graminaceous species. In the present report, we showed that OsYSL9 localizes mainly to the plasma membrane and transports both iron(II)-nicotianamine and iron(III)-deoxymugineic acid into the cell. Expression of OsYSL9 was induced in the roots but repressed in the nonjuvenile leaves in response to iron deficiency. In iron-deficient roots, OsYSL9 was induced in the vascular cylinder but not in epidermal cells. Although OsYSL9-knockdown plants did not show a growth defect under iron-sufficient conditions, these plants were more sensitive to iron deficiency in the nonjuvenile stage compared with non-transgenic plants. At the grain-filling stage, OsYSL9 expression was strongly and transiently induced in the scutellum of the embryo and in endosperm cells surrounding the embryo. The iron concentration was decreased in embryos of OsYSL9-knockdown plants but was increased in residual parts of brown seeds. These results suggested that OsYSL9 is involved in iron translocation within plant parts and particularly iron translocation from endosperm to embryo in developing seeds.

     

Effect of iron plaque outside roots on nutrient uptake by rice (Oryza sativa L.): Phosphorus uptake

Under anaerobic conditions, ferric hydroxide deposits on the surface of rice roots have been shown to affect the uptake of some nutrients. In the present experiment, different amount of this iron plaque were induced on the roots of rice (Oryza sativa L. cv. TZ88-145) by supplying different Fe(OH)₃ concentrations in nutrient solutions, and the effect of the iron plaque on phosphorus uptake was investigated. Results showed that 1) iron plaque adsorbed phosphorus from the growth medium, and that the amount of phosphorus adsorbed by the plaque was correlated with the amount of plaque; 2) the phosphorus concentration in the shoot increased by up to 72% after 72 h at concentration of Fe(OH)₃ in the nutrient solution from 0 to 30 mg Fe/L, corresponding with amounts of iron plaque from 0.2 to 24.5 mg g^-1 (root d. wt); 3) the phosphorus concentration in the shoots of rice with iron plaque was higher than that without iron plaque though the concentration in the shoot decreased when Fe(OH)₃ was added at 50 mg Fe/L producing 28.3 mg g^-1 (root d. wt) of plaque; and 4) the phosphorus concentrations in Fe-deficient and Fe-sufficient rice plants with iron plaque were the same, although phytosiderophores were released from the Fe-deficient roots. The phytosiderophores evidently did not mobilise phosphorus adsorbed on plaque. The results suggest that iron plaque on rice plant roots might be considered a phosphorus reservoir. This revised version was published online in June 2006 with corrections to the Cover Date.     

Activation of rice Yellow Stripe1-Like 16 (OsYSL16) enhances iron efficiency

Graminaceous plants release ferric-chelating phytosiderophores that bind to iron. These ferric-phytosiderophore complexes are transported across the plasma membrane by a protein produced from Yellow Stripe 1 (YS1). Here, we report the characterization of OsYSL16, one of the YS1-like genes in rice. Real-time analysis revealed that this gene was constitutively expressed irrespective of metal status. Promoter fusions of OsYSL16 to β-glucuronidase (GUS) showed that OsYSL16 was highly expressed in the vascular tissues of the root, leaf, and spikelet, and in leaf mesophyll cells. The OsYSL16-green fluorescence protein (GFP) fusion protein was localized to the plasma membrane. From a pool of rice T-DNA insertional lines, we identified two independent activation-tagging mutants in OsYSL16. On an Fe-deficient medium, those mutants retained relatively high chlorophyll concentrations compared with the wild-type (WT) controls, indicating that they are more tolerant to a lack of iron. The Fe concentration in shoots was also higher in the OsYSL16 activation lines than in the WT. During germination, the rate of Fe-utilization from the seeds was higher in the OsYSL16 activation lines than in the WT seeds. Our results suggest that the function of OsYSL16 in Fe-homeostasis is to enable distribution of iron within a plant.     

Response of rice (Oryza sativa) with root surface iron plaque under aluminium stress

BACKGROUND AND AIMS: Rice (Oryza sativa) is an aquatic plant with a characteristic of forming iron plaque on its root surfaces. It is considered to be the most Al-tolerant species among the cereal crops. The objective of this study was to determine the effects of root surface iron plaque on Al translocation, accumulation and the change of physiological responses under Al stress in rice in the presence of iron plaque. METHODS: The japonica variety rice, Koshihikari, was used in this study and was grown hydroponically in a growth chamber. Iron plaque was induced by exposing the rice roots to 30 mg L(-1) ferrous iron either as Fe(II)-EDTA in nutrient solution (6 d, Method I) or as FeSO(4) in water solution (12 h, Method II). Organic acid in root exudates was retained in the anion-exchange resin and eluted with 2 m HCl, then analysed by high-performance liquid chromatography (HPLC) after proper pre-treatment. Fe and Al in iron plaque were extracted with DCB (dithionite-citrate-bicarbonate) solution. KEY RESULTS AND CONCLUSIONS: Both methods (I and II) could induce the formation of iron plaque on rice root surfaces. The amounts of DCB-extractable Fe and Al on root surfaces were much higher in the presence of iron plaque than in the absence of iron plaque. Al contents in root tips were significantly decreased with iron plaque; translocation of Al from roots to shoots was significantly reduced with iron plaque. Al-induced secretion of citrate was observed and iron plaque could greatly depress this citrate secretion. These results suggested that iron plaque on rice root surfaces can be a sink to sequester Al onto the root surfaces and Fe ions can pre-saturate Al-binding sites in root tips, which protects the rice root tips from suffering Al stress to a certain extent.     

The impact of iron plaque on La and Nd uptake and translocation in rice (Oryza sativa L.)

The effect of root surface iron plaque formation on the uptake, transfer and accumulation of La and Nd in the rice root system was evaluated by using solution cultures. The results showed that La and Nd pollution stress inhibit formation of rice root surface iron plaques. The amount of La and Nd absorbed by the rice root surface iron plaque rose with the increase of La and Nd solution concentrations. Iron plaque formation on the rice root surface significantly decreases the La and Nd concentrations in rice roots and shoots. At growth solution La concentrations of 0.1, 0.5, and 1.0 mmol.L^? 1, concentrations of La in rice roots with induced iron plaques decreased by 17.1%, 37.4%, and 31.2%, respectively, and concentrations of La in rice shoots decreased by 43.9%, 60.6%, and 27.0%, respectively, when compared to plants with non-induced iron plaques. Also, with Nd solution concentrations of 0.1, 0.5, and 1.0 mmol.L^? 1, the Nd concentrations in rice roots and shoots of plants with induced iron plaques decreased by 21.0–31.7% and 22.7–47.5%, respectively when compared to plants with non-induced iron plaques. Iron plaque formation on the rice root surface affects the accumulation and transfer of La and Nd in rice roots. Accumulation of La and Nd was greater in rice roots than in rice shoots regardless of whether the plants had induced or non-induced iron plaques. Transfer coefficients of iron plague on rice root surface and root system under La treatments were both higher than those under Nd treatment. For rice roots and iron plaques on the root surface, the enrichment coefficient in the La treatment group was less than that in the Nd treatment group, while for rice shoots, the enrichment coefficient in the La treatment group was greater than that in the Nd treatment group. Clearly, the mechanisms governing the effect of iron plaque on La and Nd uptake and transfer in the rice root system are rather complicated.     

Iron nutrition affects cadmium accumulation and toxicity in rice plants

The effect of iron (Fe) nutrition on cadmium (Cd) toxicity and accumulation in rice plants was studied using a hydroponic system. The inhibitory effect of Cd on plant growth and chlorophyll content (SPAD value) was dependent on Fe level and the genotype. Malondialdehyde (MDA) content in leaves and roots was not much affected by an increased Cd stress at 0.171 mg l⁻¹ Fe, but it showed a rapid increase when the plants were exposed to moderate (1.89 mg l⁻¹) and high (16.8 mg l⁻¹) Fe levels. High Fe nutrition caused a marked reduction in Cd content in both leaves and roots. Fe content in plants was lower at high Cd (5.0 μM) stress than at low Cd (<1.0 μM) stress. Cd stress increased both superoxide dismutase (SOD) and peroxidase (POD) activities at low and moderate Fe levels. However, with high Fe level, it increased the POD activity, but reduced the SOD activity. Our results substantiate the hypothesis that cell membrane-bound iron transporter (carrier) involved in high-affinity iron transport systems can also transport Cd, and both these ions may compete for this common carrier. The study further showed that there were significant correlations between MDA and Fe contents in leaves and roots of rice plants. It is suggested that the occurrence of oxidative stress in plants exposed to Cd stress is mediated by Fe nutrition. The present results also show that Cd stress affects the uptake of Cu and Zn.     

Zea mays Fe deficiency‐related 4 (ZmFDR4) functions as an iron transporter in the plastids of monocots

Iron (Fe)‐homeostasis in the plastids is closely associated with Fe transport proteins that prevent Fe from occurring in its toxic free ionic forms. However, the number of known protein families related to Fe transport in the plastids (about five) and the function of iron in non‐green plastids is limited. In the present study, we report the functional characterization of Zea mays Fe deficiency‐related 4 (ZmFDR4), which was isolated from a differentially expressed clone of a cDNA library of Fe deficiency‐induced maize roots. ZmFDR4 is homologous to the bacterial FliP superfamily, coexisted in both algae and terrestrial plants, and capable of restoring the normal growth of the yeast mutant fet3fet4, which possesses defective Fe uptake systems. ZmFDR4 mRNA is ubiquitous in maize and is inducible by iron deficiency in wheat. Transient expression of the 35S:ZmFDR4–eGFP fusion protein in rice protoplasts indicated that ZmFDR4 maybe localizes to the plastids envelope and thylakoid. In 35S:c‐Myc‐ZmFDR4 transgenic tobacco, immunohistochemistry and immunoblotting confirmed that ZmFDR4 is targeted to both the chloroplast envelope and thylakoid. Meanwhile, ultrastructure analysis indicates that ZmFDR4 promotes the density of plastids and accumulation of starch grains. Moreover, Bathophenanthroline disulfonate (BPDS) colorimetry and inductively coupled plasma mass spectrometry (ICP‐MS) indicate that ZmFDR4 is related to Fe uptake by plastids and increases seed Fe content. Finally, 35S:c‐Myc‐ZmFDR4 transgenic tobacco show enhanced photosynthetic efficiency. Therefore, the results of the present study demonstrate that ZmFDR4 functions as an iron transporter in monocot plastids and provide insight into the process of Fe uptake by plastids.     

Iron plaque enhances phosphorus uptake by rice (Oryza sativa) growing under varying phosphorus and iron concentrations

Both solution culture and pot experiments were performed to investigate (a) the effects of external Fe (II) concentrations and forms on the formation of iron plaque on the roots of rice (Oryza sativa) and subsequent P adsorption on iron plaque and shoot P concentrations and (b) the effects of soil moisture regimes on the formation of iron plaque and P adsorption on root surfaces and P accumulation in shoots. The results showed that iron plaque was significantly increased with increasing Fe²⁺ concentrations in the solution culture. The amounts of P adsorbed on the iron plaque were increased significantly with external Fe²⁺ concentrations. Although shoot P concentration was not significantly affected by Fe²⁺ treatment after incubation for 2 days, it was significantly increased in the Fe‐treated plants compared with Fe‐deprived ones after incubation for 4 days. Soil culture experiment showed that the formation of iron plaque on root surfaces was promoted by exogenous iron, with greater amount of iron plaque being formed by addition of ferric hydroxide than of ferric oxide. Phosphorus adsorption on iron plaque also increased with the addition of iron oxides, and increasing soil P increased the amounts of P associated with the iron plaque and shoot P concentration. The amounts of iron plaque were almost sixfold higher under flooding condition than under field capacity condition. Plants pretreated under flooding condition generally had higher shoot P concentrations when they were transplanted to solutions with varying P levels, and this was most pronounced in the treatment with highest solution P concentration. The results suggest that iron plaque acts as a nutrient reservoir for phosphorus in the rhizosphere and helps enhance P acquisition by rice.