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Rice (Oryza sativa L.) grain is a major dietary source of cadmium (Cd), which is toxic to humans, but no practical technique exists to substantially reduce Cd contamination. Carbon ion-beam irradiation produced three rice mutants with <0.05 mg Cd⋅kg(-1) in the grain compared with a mean of 1.73 mg Cd⋅kg(-1) in the parent, Koshihikari. We identified the gene responsible for reduced Cd uptake and developed a strategy for marker-assisted selection of low-Cd cultivars. Sequence analysis revealed that these mutants have different mutations of the same gene (OsNRAMP5), which encodes a natural resistance-associated macrophage protein. Functional analysis revealed that the defective transporter protein encoded by the mutant osnramp5 greatly decreases Cd uptake by roots, resulting in decreased Cd in the straw and grain. In addition, we developed DNA markers to facilitate marker-assisted selection of cultivars carrying osnramp5. When grown in Cd-contaminated paddy fields, the mutants have nearly undetectable Cd in their grains and exhibit no agriculturally or economically adverse traits. Because mutants produced by ion-beam radiation are not transgenic plants, they are likely to be accepted by consumers and thus represent a practical choice for rice production worldwide.

Heavy metals are transported to rice grains via the phloem. In rice nodes, the diffuse vascular bundles (DVBs), which enclose the enlarged elliptical vascular bundles (EVBs), are connected to the panicle and have a morphological feature that facilitates xylem-to-phloem transfer. To find a mechanism for restricting cadmium (Cd) transport into grains, the distribution of Cd, zinc (Zn), manganese (Mn), and sulphur (S) around the vascular bundles in node I (the node beneath the panicle) of Oryza sativa 'Koshihikari' were compared 1 week after heading. Elemental maps of Cd, Zn, Mn, and S in the vascular bundles of node I were obtained by synchrotron micro-X-ray fluorescence spectrometry and electron probe microanalysis. In addition, Cd K-edge microfocused X-ray absorption near-edge structure analyses were used to identify the elements co-ordinated with Cd. Both Cd and S were mainly distributed in the xylem of the EVB and in the parenchyma cell bridge (PCB) surrounding the EVB. Zn accumulated in the PCB, and Mn accumulated around the protoxylem of the EVB. Cd was co-ordinated mainly with S in the xylem of the EVB, but with both S and O in the phloem of the EVB and in the PCB. The EVB in the node retarded horizontal transport of Cd toward the DVB. By contrast, Zn was first stored in the PCB and then efficiently transferred toward the DVB. Our results provide evidence that transport of Cd, Zn, and Mn is differentially controlled in rice nodes, where vascular bundles are functionally interconnected.

The effects of water management in rice paddies on the levels of cadmium (Cd) and arsenic (As) in Japanese rice grains were tested. In order to reduce the Cd concentration in rice grains, flooding for 3 weeks before and after heading was most effective, but this treatment increased As concentration considerably. Aerobic treatment was effective in reducing As concentration in rice grains, but increased Cd concentration markedly. In the pot experiment, flooding treatment after heading was more effective than flooding treatment before heading in reducing both Cd and As concentrations in rice grains. The concentration of dimethylarsinic acid (DMA) in rice grains was very low under aerobic conditions, but increased in the continuous-flooding treatment. In the field experiment, the grain As concentration in the case of flooding for 3 weeks before and after heading was higher than that in the case of intermittent irrigation. The ratios of DMA to the total As concentration were 3-52% in the pot experiment and 7-13% in the field experiment.

Cadmium (Cd) is a heavy metal toxic to humans and the accumulation of Cd in the rice grain is a major agricultural problem, particularly in Asia. The role of the iron transporter OsNRAMP1 in Cd uptake and transport in rice was investigated here. An OsNRAMP1:GFP fusion protein was localized to the plasma membrane in onion epidermal cells. The growth of yeast expressing OsNRAMP1 was impaired in the presence of Cd compared with yeast transformed with an empty vector. Moreover, the Cd content of OsNRAMP1-expressing yeast exceeded that of the vector control. The expression of OsNRAMP1 in the roots was higher in a high Cd-accumulating cultivar (Habataki) than a low Cd-accumulating cultivar (Sasanishiki) regardless of the presence of Cd, and the amino acid sequence of OsNRAMP1 showed 100% identity between Sasanishiki and Habataki. Over-expression of OsNRAMP1 in rice increased Cd accumulation in the leaves. These results suggest that OsNRAMP1 participates in cellular Cd uptake and Cd transport within plants, and the higher expression of OsNRAMP1 in the roots could lead to an increase in Cd accumulation in the shoots. Our results indicated that OsNRAMP1 is an important protein in high-level Cd accumulation in rice.

Background: A simple and quick on-site test for trace levels of cadmium (Cd) in food is needed because of the human toxicity of this heavy metal. We developed an immunochromatography kit which uses the antigen-antibody complex reaction between the Cd-ethylenediaminetetraacetic acid (Cd-EDTA) complex and an anti-Cd-EDTA antibody. We previously reported the successful use of this kit to determine Cd concentrations in brown rice with respect to the international standard: 0.4 mg kg⁻¹. Here, we measured, using this immunochromatography kit, Cd concentrations in crops with lower international standards than rice., Results: Cadmium extracted with 0.1 mol L⁻¹ HCl from wheat grain and fresh eggplant was purified sufficiently using an ion-exchange column treatment. Appropriate HCl extraction rates and dilution rates for the column eluate were selected; Cd concentrations in wheat grain and fresh eggplant were determined successfully by immunochromatography with respect to the international standards of 0.2 mg kg⁻¹ and 0.05 mg kg⁻¹ fresh weight, respectively., Conclusion: Approximate Cd concentrations in wheat grain and fresh eggplant can be monitored easily and quickly by this method at locations where facilities for acid digestion and precision analysis are not available., (Copyright © 2011 Society of Chemical Industry.)

Solanum torvum Sw. cv. Torubamubiga (TB) is a low cadmium (Cd)-accumulating plant. To elucidate the molecular mechanisms of the Cd acclimation process in TB roots, transcriptional regulation was analysed in response to mild Cd treatment: 0.1 muM CdCl(2) in hydroponic solution. A unigene set consisting of 6296 unigene sequences was constructed from 18 816 TB cDNAs. The distribution of functional categories was similar to tomato, while 330 unigenes were suggested to be TB specific. For expression profiling, the SuperSAGE method was adapted for use with Illumina sequencing technology. Expression tag libraries were constructed from Cd-treated (for 3 h, 1 d, and 3 d) and untreated roots, and 34 269 species of independent tags were collected. Moreover, 6237 tags were ascribed to the TB or eggplant (aubergine) unigene sequences. Time-course changes were examined, and 2049 up- and 2022 down-regulated tags were identified. Although no tags annotated to metal transporter genes were significantly regulated, a tag annotated to AtFRD3, a xylem-loading citrate transporter, was down-regulated. In addition to induction of heavy metal chaperone proteins, antioxidative and sulphur-assimilating enzymes were induced, confirming that oxidative stress developed even using a mild Cd concentration. Rapid repression of dehydration-related transcription factors and aquaporin isoforms suggests that dehydration stress is a potential constituent of Cd-induced biochemical impediments. These transcriptional changes were also confirmed by real-time reverse transcription-PCR. Further additions of TB unigene sequences and functional analysis of the regulated tags will reveal the molecular basis of the Cd acclimation process, including the low Cd-accumulating characteristics of TB.

Rice consumption is a major source of cadmium and arsenic for the population of Asia. We investigated the effects of water management in rice paddy on levels of cadmium and arsenic in Japanese rice grains. Flooding increased arsenic concentrations in rice grains, whereas aerobic treatment increased the concentration of cadmium. Flooding for 3 weeks before and after heading was most effective in reducing grain cadmium concentrations, but this treatment increased the arsenic concentration considerably, whereas aerobic treatment during the same period was effective in reducing arsenic concentrations but increased the cadmium concentration markedly. Flooding treatment after heading was found to be more effective than flooding treatment before heading in reducing rice grain cadmium without a concomitant increase in total arsenic levels, although it increased inorganic arsenic levels. Concentrations of dimethylarsinic acid (DMA) in grain were very low under aerobic conditions but increased under flooded conditions. DMA accounted for 3-52% of the total arsenic concentration in grain grown in soil with a lower arsenic concentration and 10-80% in soil with a higher arsenic concentration. A possible explanation for the accumulation of DMA in rice grains is that DMA translocates from shoots/roots to the grains more readily than does inorganic arsenic.

In accordance with a new international standard set by the Codex Alimentarius Commission for cadmium (Cd) contentin rice grain, Japan must perform large-scale remediation of paddy fields polluted with lowto moderate levels of Cd. Phytoextraction using hyperaccumulating wild plants has been proposed as a low-cost, environmentally friendly restoration technology. However, because of difficulties with sowing, weed and disease control, and harvesting, hyperaccumulators may not be suitable for large-scale phytoextraction in polluted paddy fields. Here, we demonstrated phytoextraction using Indica-type rice cultivars capable of accumulating Cd at high levels. Phytoextraction with the Indica rice Chokoukoku grown for 2 years without irrigation after drainage removed 883 g Cd ha(-1), reduced the total soil Cd content by 38%, and reduced the grain Cd content in subsequently grown Japonica food rice by 47% without decreasing yield. The results suggest that phytoextraction with Chokoukoku can remove Cd from paddy fields polluted with low to moderate levels of Cd and reduce the grain Cd concentration of Japonica food rice cultivars to below the Codex standard within a reasonable time frame. This approach will help reduce the risk of Cd contamination of rice from paddy fields.

Physiological properties involved in divergent cadmium (Cd) accumulation among rice genotypes were characterized using the indica cultivar 'Habataki' (high Cd in grains) and the japonica cultivar 'Sasanishiki' (low Cd in grains). Time-dependence and concentration-dependence of symplastic Cd absorption in roots were revealed not to be responsible for the different Cd accumulation between the two cultivars because root Cd uptake was not greater in the Cd-accumulating cultivar 'Habataki' compared with 'Sasanishiki'. On the other hand, rapid and greater root-to-shoot Cd translocation was observed in 'Habataki', which could be mediated by higher abilities in xylem loading of Cd and transpiration rate as a driving force. To verify whether different abilities in xylem-mediated shoot-to-root translocation generally account for the genotypic variation in shoot Cd accumulation in rice, the world rice core collection, consisting of 69 accessions which covers the genetic diversity of almost 32,000 accessions of cultivated rice, was used. The results showed strong correlation between Cd levels in xylem sap and shoots and grains among the 69 rice accessions. Overall, the results presented in this study revealed that the root-to-shoot Cd translocation via the xylem is the major and common physiological process determining the Cd accumulation level in shoots and grains of rice plants.

We determined the natural Cd balance in a cultivated rice paddy field in Japan. The main sources of Cd in the non-polluted paddy field were phosphorus fertilizer and precipitation (annual input of Cd, 2,000 mg ha(-1) and 1,020 mg ha(-1), respectively). These sources account for 95% of the total input of Cd (3,192 mg ha(-1)). The actual increase of Cd in the soil was 0.0016 mg kg(-1), we thus consider Cd in soil increases only slightly as a result of rice culture. This study indicates that it is difficult to reduce Cd loading by irrigation water treatment in a non-polluted paddy field. This further indicates that once a field is polluted by Cd it is difficult to decrease the accumulated Cd by the ordinary cultivation of rice plants., (Copyright (c) IWA Publishing 2008.)