INVESTIGATION OF SHORT-TERM CHEMICAL CHANGE IN STABLE RUTHENIUM ADDED TO RAINWATER THROUGH X-RAY ABSORPTION FINE STRUCTURE ANALYSIS.

Radioactive ruthenium may be accidentally released from spent nuclear fuel reprocessing plants to the surrounding environment. To obtain basic information regarding the effect of radioactive ruthenium on the environment, we investigated changes in the source form of stable ruthenium added to rainwater through X-ray absorption fine structure analysis. Ruthenium tetroxide (RuO4), ruthenium dioxide (RuO2), ruthenium nitrosyl nitrate (Ru(NO)(NO3)3) and ruthenium chloride (RuCl3) were employed as test sources. The Ru K-edge X-ray absorption near edge structure spectra acquired immediately after addition to ultrapure water differed among various sources. With RuO4 addition, a black deposit was observed 1 d after addition, and the spectrum was similar to that of RuO2. For RuO2, Ru(NO)(NO3)3 and RuCl3, no variances were observed between the spectra acquired immediately after addition and 1 d after addition. These results indicate that the chemical forms of RuO2, Ru(NO)(NO3)3 and RuCl3 did not change within a 1-d span in rainwater.

SOIL-SOIL SOLUTION DISTRIBUTION COEFFICIENT OF RADIOIODINE IN SURFACE SOILS AROUND SPENT NUCLEAR FUEL REPROCESSING PLANT IN ROKKASHO, JAPAN.

The soil-soil solution distribution coefficient (Kd) of radioiodine in soil samples with various total carbon (TC) contents was measured in a batch sorption experiment using 125I tracer spiked as I-. The log values of Kd-125I and TC concentration in low-TC soils (< 10g kg-1) were positively correlated, whereas those of Kd-125I in TC rich soils (> 10 g kg-1) and dissolved organic carbon (DOC) in liquid phase were negatively correlated. The proportion of 125I in the < 3 kDa fraction in the liquid phase is negatively correlated with the log of DOC, implying that 125I is primarily combined with high-molecular-weight organic matter in soil solutions rich in DOC. The results suggest that Kd-125I in soil with high soil organic material (SOM) content is governed by DOC via the combination of 125I and DOC. In contrast, Kd-125I in soils with a low SOM content was governed by SOM because the anion exchange capacity of SOM was vital for the sorption of 125I-.

Visualising spatio-temporal distributions of assimilated carbon translocation and release in root systems of leguminous plants.

The release of rhizodeposits differs depending on the root position and is closely related to the assimilated carbon (C) supply. Therefore, quantifying the C partitioning over a short period may provide crucial information for clarifying root-soil carbon metabolism. A non-invasive method for visualising the translocation of recently assimilated C into the root system inside the rhizobox was established using 11CO2 labelling and the positron-emitting tracer imaging system. The spatial distribution of recent 11C-photoassimilates translocated and released in the root system and soil were visualised for white lupin (Lupinus albus) and soybean (Glycine max). The inputs of the recently assimilated C in the entire root that were released into the soil were approximately 0.3%-2.9% for white lupin within 90 min and 0.9%-2.3% for soybean within 65 min, with no significant differences between the two plant species; however, the recently assimilated C of lupin was released at high concentrations in specific areas (hotspots), whereas that of soybean was released uniformly in the soil. Our method enabled the quantification of the spatial C allocations in roots and soil, which may help to elucidate the relationship between C metabolism and nutrient cycling at specific locations of the root-soil system in response to environmental conditions over relatively short periods.

Diversity of Antibiotic Biosynthesis Gene-possessing Rhizospheric Fluorescent Pseudomonads in Japan and Their Biocontrol Efficacy.

More than 3,000 isolates of fluorescent pseudomonads have been collected from plant roots in Japan and screened for the presence of antibiotic-synthesizing genes. In total, 927 hydrogen cyanide (HCN)-, 47 2,4-diacetylphloroglucinol (PHL)-, 6 pyoluteorin (PLT)-, 14 pyrrolnitrin (PRN)-, and 8 phenazine (PHZ)-producing isolates have been detected. A cluster analysis (≥99% identity) identified 10 operational taxonomic units (OTUs) in antibiotic biosynthesis gene-possessing pseudomonads. OTU HLR (PHL, PLT, and PRN) contained four antibiotics: HCN, PHL, PLT, and PRN, while OTU RZ (PRN and PHZ) contained three: HCN, PRN, and PHZ. OTU H1, H2, H3, H4, H5, H6, and H7 (PHL1-7) contained two antibiotics: HCN and PHL, while OTU H8 (PHL8) contained one: PHL. Isolates belonging to OTU HLR and RZ suppressed damping-off disease in cabbage seedlings caused by Rhizoctonia solani. Effective strains belonging to OTU HLR and RZ were related to Pseudomonas protegens and Pseudomonas chlororaphis, respectively. Antibiotic biosynthesis gene-possessing fluorescent pseudomonads are distributed among different geographical sites in Japan and plant species.

Effects of organic amendments on the natural attenuation of radiocesium transferability in grassland soils with high potassium fertility.

Organic amendments affect the behavior of radiocesium in soil-plant systems in a complex way; they can inhibit radiocesium fixation by clay minerals by blocking selective sorption sites, whereas K supplied to the soil solution by amendments can reduce Cs uptake by plant roots. Here, we investigated the influence of inorganic and organic amendments on the transferability of radiocesium from soil to grass seedling in a humus-rich Andosol with high exchangeable K content. Soil samples were spiked with a137Cs tracer, treated with N, N-P-K, compost (cattle manure using rice straw), or no amendment (control soil), and subjected to repeated two-week wetting and air-drying treatments for one year in an artificial climate chamber. Small-scale cultivations of orchard grass were performed four times during the experimental period to assess temporal changes of availability of 137Cs in the soils. The 137Cs transfer factor (TF), defined as the 137Cs concentration in the plant divided by that in the soil, decreased with time in the control soil. The soil treated with compost showed higher TFs than the control soil in each cultivation and a slower attenuation of 137Cs transferability. By comparing the extractability of 137Cs, NH4+, and K+ with the observed TFs, we show that K released from the compost was not effective in reducing root uptake of 137Cs, but enhanced 137Cs desorption from the soil under K-rich conditions. This result suggests that organic amendment is ineffective in reducing root uptake of radiocesium under high exchangeable K concentrations, and may instead enhance the long-term availability of radiocesium in soils.

Assessment of bacterial communities of black soybean grown in fields.

Since the domestication of soybean (Glycine max) about 4,500 years ago, thousands of local cultivars have been developed around the world. In Japan, black soybeans grown in the mountainous region of central Kyoto and Hyogo prefectures, called the Tamba region, are well known for large seeds and palatability. The yields of black soybean in the Tamba region of Kyoto have decreased during the past few decades, and the involvement of rhizosphere microbes in the yield decline has been suggested. We analyzed bacterial communities of the soybean rhizosphere on 7 farms managed under different strategies. Non-metric multidimensional scaling showed shifts of bacterial communities from bulk to rhizosphere soil and the difference among the farms. The relative abundance of the Proteobacteria and Firmicutes was higher in rhizosphere soil than in bulk soil, whereas that of the Acidobacteria was higher in bulk soil. To clarify the possible relationship between bacterial communities and soybean growth, we used ConfeitoGUIplus software (version 1.2.0), based on the Confeito algorithm, which is designed to detect highly interconnected modules in a correlation network by using a unique inter-modular index with network density. One module was extracted from the rhizosphere soil community and two from bulk soil communities, suggesting the involvement of these bacteria in soybean growth.

Soil-soil solution distribution coefficient of soil organic matter is a key factor for that of radioiodide in surface and subsurface soils.

We investigated the vertical distribution of the soil-soil-solution distribution coefficients (Kd) of 125I, 137Cs, and 85Sr in organic-rich surface soil and organic-poor subsurface soil of a pasture and an urban forest near a spent-nuclear-fuel reprocessing plant in Rokkasho, Japan. Kd of 137Cs was highly correlated with water-extractable K+. Kd of 85Sr was highly correlated with water-extractable Ca2+ and SOC. Kd of 125I- was low in organic-rich surface soil, high slightly below the surface, and lowest in the deepest soil. This kinked distribution pattern differed from the gradual decrease of the other radionuclides. The thickness of the high-125I-Kd middle layer (i.e., with high radioiodide retention ability) differed between sites. Kd of 125I- was significantly correlated with Kd of soil organic carbon. Our results also showed that the layer thickness is controlled by the ratio of Kd-OC between surface and subsurface soils. This finding suggests that the addition of SOC might prevent further radioiodide migration down the soil profile. As far as we know, this is the first report to show a strong correlation of a soil characteristic with Kd of 125I-. Further study is needed to clarify how radioiodide is retained and migrates in soil.

Determination of Iodide, Iodate and Total Iodine in Natural Water Samples by HPLC with Amperometric and Spectrophotometric Detection, and Off-line UV Irradiation.

We developed a rapid, simple method for the iodine speciation analysis of water and applied it to natural water samples. Simultaneous determinations of I(-) and IO3(-) were achieved with an HPLC system with amperometric detection for I(-) and spectrophotometric detection after a postcolumn reaction for IO3(-). We determined the I(-) and IO3(-) concentrations in 20-µL water samples within 10 min. Total I concentrations in water samples were determined after the decomposition of organics by off-line UV irradiation for 30 min, followed by reduction to I(-). The analytical conditions were optimized by using test solutions rich in organic matter extracted from soils. We tested the new method with samples of groundwater, spring water, precipitation, soil percolate, stream water, and seawater as well as solutions extracted from soil. The method worked well, although the concentrations of some I species were below detection. This method is suitable for routine speciation analysis, which is important for studies of I behavior in the environment.

A putative multicopper oxidase, IoxA, is involved in iodide oxidation by Roseovarius sp. strain A-2.

Roseovarius sp. strain A-2 is an aerobic heterotrophic bacterium with a capacity for oxidizing iodide ion (I(-)) to form molecular iodine (I2). In this study, iodide-oxidizing enzyme of strain A-2 was characterized. The enzyme was an extracellular protein, and Cu(2+) ion significantly enhanced the enzyme activity in the culture supernatant. When iodide was used as the substrate, the crude enzyme showed Km and Vmax values of 4.78 mM and 25.1 U mg(-1), respectively. The enzyme was inhibited by NaN3, EDTA, KCN, and o-phenanthroline, and also had significant activities toward p-phenylenediamine and hydroquinone. Tandem mass spectrometric analysis of an active band excised from SDS-PAGE gel revealed that at least two proteins are involved in the enzyme. One of these proteins was closely related with IoxA, a multicopper oxidase previously found as a component of iodide-oxidizing enzyme of Alphaproteobacterium strain Q-1. Furthermore, a terrestrial bacterium Rhodanobacter denitrificans 116-2, which possesses an ioxA-like gene in its genome, was found to oxidize iodide. These results suggest that IoxA catalyzes the oxidation of iodide in phylogenetically distinct bacteria.

Effect of phosphorus levels on the protein profiles of secreted protein and root surface protein of rice.

Plant roots are complicated organs that absorb water and nutrients from the soil. Roots also play an essential role in protecting plants from attack by soil pathogens and develop a beneficial role with some soil microorganisms. Plant-derived rhizosphere proteins (e.g., root secretory proteins and root surface binding proteins) are considered to play important roles in developing mutual relationships in the rhizosphere. In the rhizosphere, where plant roots meet the surrounding environment, it has been suggested that root secretory protein and root surface binding protein are important factors. Furthermore, it is not known how the physiological status of the plant affects the profile of these proteins. In this study, rice plants were grown aseptically, with or without phosphorus nutrition, and proteins were obtained from root bathing solution (designated as root secretory proteins) and obtained using 0.2 M CaCl2 solution (designated as root surface binding proteins). The total number of identified proteins in the root bathing solution was 458, and the number of root surface binding proteins was 256. More than half of the proteins were observed in both fractions. Most of the proteins were categorized as either having signal peptides or no membrane transport helix sites. The functional categorization suggested that most of the proteins seemed to have secretory pathways and were involved in defense/disease-related functions. These characteristics seem to be unique to rhizosphere proteins, and the latter might be part of the plants strategy to defeat pathogens in the soil. The low phosphorus treatment significantly increased the number of pathogenesis-related proteins in the root secretory proteins, whereas the change was small in the case of the root surface binding proteins. The results suggested that the roots are actively and selectively secreting protein into the rhizosphere.

Metagenomic analysis of the rhizosphere soil microbiome with respect to phytic acid utilization.

While phytic acid is a major form of organic phosphate in many soils, plant utilization of phytic acid is normally limited; however, culture trials of Lotus japonicus using experimental field soil that had been managed without phosphate fertilizer for over 90 years showed significant usage of phytic acid applied to soil for growth and flowering and differences in the degree of growth, even in the same culture pot. To understand the key metabolic processes involved in soil phytic acid utilization, we analyzed rhizosphere soil microbial communities using molecular ecological approaches. Although molecular fingerprint analysis revealed changes in the rhizosphere soil microbial communities from bulk soil microbial community, no clear relationship between the microbiome composition and flowering status that might be related to phytic acid utilization of L. japonicus could be determined. However, metagenomic analysis revealed changes in the relative abundance of the classes Bacteroidetes, Betaproteobacteria, Chlorobi, Dehalococcoidetes and Methanobacteria, which include strains that potentially promote plant growth and phytic acid utilization, and some gene clusters relating to phytic acid utilization, such as alkaline phosphatase and citrate synthase, with the phytic acid utilization status of the plant. This study highlights phylogenetic and metabolic features of the microbial community of the L. japonicus rhizosphere and provides a basic understanding of how rhizosphere microbial communities affect the phytic acid status in soil.

Analysis of diversity of diazotrophic bacteria associated with the rhizosphere of a tropical Arbor, Melastoma malabathricum L.

The diversity of diazotrophic bacteria in the rhizosphere of Melastoma malabathricum L. was investigated by cloning-sequencing of the nifH gene directly amplified from DNA extracted from soil. Samples were obtained from the rhizosphere and bulk soil of M. malabathricum growing in three different soil types (acid sulfate, peat and sandy clay soils) located very close to each other in south Kalimantan, Indonesia. Six clone libraries were constructed, generated from bulk and rhizosphere soil samples, and 300 nifH clones were produced, then assembled into 29 operational taxonomic units (OTUs) based on percent identity values. Our results suggested that nifH gene diversity is mainly dependent on soil properties, and did not differ remarkably between the rhizosphere and bulk soil of M. malabathricum except in acid sulfate soil. In acid sulfate soil, as the Shannon diversity index was lower in rhizosphere than in bulk soil, it is suggested that particular bacterial species might accumulate in the rhizosphere.

Plant growth promotion abilities and microscale bacterial dynamics in the rhizosphere of Lupin analysed by phytate utilization ability.

In the rhizosphere, phosphorus (P) levels are low because of P uptake into the roots. Rhizobacteria live on carbon (C) exuded from roots, and may contribute to plant nutrition by liberating P from organic compounds such as phytates. We isolated over 300 phytate (Na-inositol hexa-phosphate; Na-IHP)-utilizing bacterial strains from the rhizosheath and the rhizoplane of Lupinus albus (L.). Almost all of the isolates were classified as Burkholderia based on 16S rDNA sequence analysis. Rhizosheath isolates cultured with Na-IHP as the only source of C and P showed lower P uptake at the same extracellular phytase activity than rhizoplane strains, suggesting that bacteria from the rhizosheath utilized phytate as a C source. Many isolates also utilized insoluble phytate (Al-IHP and/or Fe-IHP). In co-culture with Lotus japonicus seedlings, some isolates promoted plant growth significantly.