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Positron imaging technique is a new technology that enables to observe the materials transportation in living things using RI tracer labeled with a positron-emitting nuclide. In the positron imaging, a pair of annihilation gamma rays, generated by pair annihilation of a positron with an electron, are detected coincidently. In medical, positron emission tomography (PET) is used in a diagnostic imaging of cancer as a powerful tool. Plant positron imaging system is using the same measurement principle with PET and is developed as a device specialized for research on plants.

More than a hundred years have passed since W. C. Rontgen found X-ray in 1895. During this period, radiation is used for not only diagnosis and treatment of disease but also elucidation of principal of life through research on human and living things. In Japan, cancer is the leading cause of the death of the people, and radiation plays an important role in the diagnosis and treatment of cancer. In this chapter, I will outline the situation of radiation use which is indispensable for research and development of the medical field and medical science which supports it, including diagnosis and treatment of cancer.

An enzymic-HPLC method was successfully applied to the simultaneous analysis of the galacturonate and neutral sugar contents of pectic substances. A mixture of seven neutral sugars that are present in oridnary pectins was eluted as one peak on a Shodex SUGAR SH1821 column, using 0.001 N sulfuric acid as the eluent; the peak was completely separated from that of galacturonate. No difference in the peak-area ratios of individual neutral sugars to glycerol (internal standard) was found among the seven; the relationship between the peak response and concentration was strictly linear throughout the entire concentration range studied. Seventeen pectic samples, including pectins, pectates, and rhamnogalacturonan fragments, were completely prehydrolyzed by Driselase, an industrial enzyme product from Irpex lacteus, and then analyzed for their constituent sugar contents. The enzymic-HPLC method was simple, accurate, and particularly useful for routine pectin analyses.

Knowledge of interspecies scaling of organ doses from experimental animals such as mice to humans is important in the preclinical evaluation of new radiopharmaceuticals. Interspecies scaling factors should be reliably determined since the dose-response relationships in mice would be translated to those in humans. To obtain reliable interspecies scaling factors of organ doses from mice to humans, absorbed fractions (AFs) are needed for sophisticated models on both mice and humans. In the present study, self-AFs for photons and electrons in the spleen, kidneys, and liver of both a mouse and humans were evaluated using Monte Carlo simulations. For the mouse and human models, voxel phantoms based on computed tomography were used. The sources were assumed to be monoenergetic in the energy range 10 keV to 4 MeV and to be uniformly distributed in the spleen, kidneys, and liver. Interspecies scaling factors were determined using the results of the self-AFs for the voxel mouse and voxel human. Consequently, interspecies scaling factors were found to be dependent upon energy emitted in the source organ. It was found that the scaling factor for the photon self-AF, which is corrected by the cube root of the organ mass, shows a similar trend as a function of energy with the scaling factor for the electron self-AF.

The real-time translocation of iron (Fe) in barley (Hordeum vulgare L. cv. Ehimehadaka no. 1) was visualized using the positron-emitting tracer (52)Fe and a positron-emitting tracer imaging system (PETIS). PETIS allowed us to monitor Fe translocation in barley non-destructively under various conditions. In all cases, (52)Fe first accumulated at the basal part of the shoot, suggesting that this region may play an important role in Fe distribution in graminaceous plants. Fe-deficient barley showed greater translocation of (52)Fe from roots to shoots than did Fe-sufficient barley, demonstrating that Fe deficiency causes enhanced (52)Fe uptake and translocation to shoots. In the dark, translocation of (52)Fe to the youngest leaf was equivalent to or higher than that under the light condition, while the translocation of (52)Fe to the older leaves was decreased, in both Fe-deficient and Fe-sufficient barley. This suggests the possibility that the mechanism and/or pathway of Fe translocation to the youngest leaf may be different from that to the older leaves. When phloem transport in the leaf was blocked by steam treatment, (52)Fe translocation from the roots to older leaves was not affected, while (52)Fe translocation to the youngest leaf was reduced, indicating that Fe is translocated to the youngest leaf via phloem in addition to xylem. We propose a novel model in which root-absorbed Fe is translocated from the basal part of the shoots and/or roots to the youngest leaf via phloem in graminaceous plants.

The radionuclide 67Cu was produced via the 68Zn(p,2p)67Cu reaction by irradiating enriched 68Zn targets with 70 MeV proton beam. Copper-67 was chemically separated from the zinc target by ion-exchange chromatography using Chelex-100 chelating ion-exchange resin. Procedure for recovery of the enriched 68Zn was developed. The target recovery yield of this method was evaluated to be more than 97%.

Deoxymugineic acid (DMA) is a member of the mugineic acid family phytosiderophores (MAs), which are natural metal chelators produced by graminaceous plants. Rice secretes DMA in response to Fe deficiency to take up Fe in the form of Fe(III)–MAs complex. In contrast with barley, the roots of which secrete MAs in response to Zn deficiency, the amount of DMA secreted by rice roots was slightly decreased under conditions of low Zn supply. There was a concomitant increase in endogenous DMA in rice shoots, suggesting that DMA plays a role in the translocation of Zn within Zn-deficient rice plants. The expression of OsNAS1 and OsNAS2 was not increased in Zn-deficient roots but that of OsNAS3 was increased in Zn-deficient roots and shoots. The expression of OsNAAT1 was also increased in Zn-deficient roots and dramatically increased in shoots; correspondingly, HPLC analysis was unable to detect nicotianamine in Zn-deficient shoots. The expression of OsDMAS1 was increased in Zn-deficient shoots. Analyses using the positron-emitting tracer imaging system (PETIS) showed that Zn-deficient rice roots absorbed less 62Zn-DMA than 62Zn2+. Importantly, supply of 62Zn-DMA rather than 62Zn2+ increased the translocation of 62Zn into the leaves of Zn-deficient plants. This was especially evident in the discrimination center (DC). These results suggest that DMA in Zn-deficient rice plants has an important role in the distribution of Zn within the plant rather than in the absorption of Zn from the soil. Electronic supplementary material The online version of this article (doi:10.1007/s11103-008-9292-x) contains supplementary material, which is available to authorized users.

Iron (Fe) deficiency is a worldwide agricultural problem on calcareous soils with low-Fe availability due to high soil pH. Rice plants use a well documented phytosiderophore-based system (Strategy II) to take up Fe from the soil and also possess a direct Fe 2+ transport system. Rice plants are extremely susceptible to low-Fe supply, however, because of low phytosiderophore secretion and low Fe 3+ reduction activity. A yeast Fe 3+ chelate-reductase gene refre1/372 , selected for better performance at high pH, was fused to the promoter of the Fe-regulated transporter, OsIRT1 , and introduced into rice plants. The transgene was expressed in response to a low-Fe nutritional status in roots of transformants. Transgenic rice plants expressing the refre1/372 gene showed higher Fe 3+ chelate-reductase activity and a higher Fe-uptake rate than vector controls under Fe-deficient conditions. Consequently, transgenic rice plants exhibited an enhanced tolerance to low-Fe availability and 7.9× the grain yield of nontransformed plants in calcareous soils. This report shows that enhancing the Fe 3+ chelate-reductase activity of rice plants that normally have low endogenous levels confers resistance to Fe deficiency.

Until now, the real-time uptake and movement of manganese (Mn), an essential plant nutrient, has not been documented in plants. In this study, the real-time translocation of Mn in barley (Hordeum vulgare L. cv. Ehimehadaka no. 1) was visualized using the positron-emitting tracer 52Mn and a positron-emitting tracer imaging system (PETIS). PETIS allowed the non-destructive monitoring of Mn translocation in barley under various conditions. In all cases, 52Mn first accumulated in the discrimination center (DC) at the basal portion of the shoot, suggesting that this region may play an important role in Mn distribution in graminaceous plants. Manganese-deficient barley showed greater translocation of 52Mn from roots to shoots than did Mn-sufficient barley, demonstrating that Mn deficiency causes enhanced Mn uptake and loading into vascular bundles. In contrast, the translocation of 52Mn from roots to shoots was suppressed in Mn-excess barley. In these plants, the uptake of Mn may be suppressed or Mn ma...

Mugineic acid family phytosiderophores (MAs) are metal chelators that are produced in graminaceous plants in response to iron (Fe) deficiency, but current evidence regarding secretion of MAs during zinc (Zn) deficiency is contradictory. Our studies using HPLC analysis showed that Zn deficiency induces the synthesis and secretion of MAs in barley plants. The levels of the HvNAS1, HvNAAT-A, HvNAAT-B, HvIDS2 and HvIDS3 transcripts, which encode the enzymes involved in the synthesis of MAs, were increased in Zn-deficient roots. Studies of the genes involved in the methionine cycle using microarray analysis showed that the transcripts of these genes were increased in both Zn-deficient and Fe-deficient barley roots, probably allowing the plant to meet its demand for methionine, a precursor in the synthesis of MAs. In addition, HvNAAT-B transcripts were detected in Zn-deficient shoots, but not in those that were deficient in Fe. Increased synthesis of MAs in Zn-deficient barley was not due to a deficiency of Fe, because Zn-deficient barley accumulated more Fe than did the control plants, ferritin transcripts were increased in Zn-deficient plants, and Zn deficiency promoted Fe transport from root to shoot. Moreover, analysis using the positron-emitting tracer imaging system (PETIS) confirmed that more 62Zn(II)-MAs than 62Zn2+ were absorbed by the roots of Zn-deficient barley plants. These data suggest that the increased biosynthesis and secretion of MAs arising from a shortage of Zn are not due to an induced Fe deficiency, and that secreted MAs are effective in absorbing Zn from the soil.

The positron emitting tracer imaging system (PETIS) and carbon-11-labeled carbon dioxide (11CO2) can be used for imaging the photosynthesis process in plant leaves. Further, 11C kinetics facilitate the estimation of the physiological function parameters of photosynthesis. PETIS measurements were performed under four light conditions for each exposure of a single leaf to 11CO2 gas. In order to estimate the rate constants of the photosynthesis parameters, the time-activity curves of the input 11CO2 gas and the leaf response were fitted to an appropriate compartmental tracer kinetic model that considers photoassimilation and sucrose export rate constants as influx and efflux, respectively. The data obtained by this method show a reasonable response with respect to the photoenvironment of the leaf, and they are important for discussing photosynthesis with regard to plant physiology and agriculture

Non-invasive real-time visualization of the accumulation of photoassimilates in the grains of an ear of wheat using [ 11 C]CO 2 and positron emitting tracer imaging system (PETIS) was studied. [ 11 C]CO 2 was supplied to the center of a fully expanded leaf of a wheat plant for an initial 10 min, and the transportation of 11 C-labeled photoassimilates into the grains of the ear was monitored for 120 min using the PETIS. Each grain was clearly identified in the obtained animation. The 11 C-labeled photoassimilates arrived at the ear from the [ 11 C]CO 2 -absorbing leaf within 53 min from the time of supplying [ 11 C]CO 2 . After that, grains appeared on the image one by one from the basal part and full images of the grains appeared within 20 min. The time course of the accumulation of photoassimilates into each grain showed a different profile. Furthermore, the PETIS data suggested that the photo-condition of the ear plays an important role in the transportation of photoassimilates in wheat. PETIS can be used to visualize the dynamics of the substances in a living plant in real time and can exhibit the time course analysis of substances, such as the transportation, distribution, and accumulation.

The photoassimilate flow in an intact plant stem was imaged in real-time and its dynamics was quantitatively described using the Positron Emitting Tracer Imaging System (PETIS). Radioactive 11CO2 was fed to a leaf of an intact broad bean (Vicia faba L.) plant, together with air containing an ambient concentration of non-radioactive carrier CO2 gas. Movies of flow of the 11C-labeled photoassimilates in the plant body were captured with PETIS. Here we demonstrate that the average flow speeds and the distribution ratios of photoassimilates in the respective nodes and internodes of the observed stem can be estimated by the transfer function analysis, one of the mathematical modeling methods. We also estimated the changes in the spatial distribution of the average flow speeds in the same stem when the fed leaf was exposed to enriched carrier CO2 gas.

The effects of anion channel blockers, 4,4′-diisothiocyanatostilbene-2,2′-disulfonic acid (DIDS) and anthracene-9-carboxylic acid (A-9-C), on the uptake and xylem transport of nitrate were investigated in intact barley (Hordeum vulgare L.) seedlings using 13NO3 − and 15NO3 − as tracers. The seedling roots were pretreated with either blocker for 1 h and then 13NO3 − or 15NO3 − was supplied to the medium. Real-time images of 13N accumulation in shoots were monitored during the first 30 min using Positron Emitting Tracer Imaging Systems (PETIS). The radioactivity in the shoots of the plants treated with either blocker was about half of that in the shoots of the control plants. Analysis of the distribution of 13N in whole seedlings using a Bioimaging Analyzer System (BAS) showed a 13N-accumulation in both shoots and roots of the control plants but reduced 13N-levels in the shoots of the plants treated with either blocker. Nitrate concentrations in the xylem sap were significantly reduced by the appli...

Water (H 2 15 O) translocation from the roots to the top of rice plants (Oryza saliva L. cv. Nipponbare) was visualized over time by a positron-emitting tracer imaging system (PETIS). H 2 15 O flow was activated 8 min after plants were exposed to bright light (1500 μmol m -2 s -1 ). When the light was subsequently removed, the flow gradually slowed and completely stopped after 12 min. In plants exposed to low light (500 μmol m -2 s -1 ), H 2 15 O flow was activated more slowly, and a higher translocation rate of H 2 15 O was observed in the same low light at the end of the next dark period. NaCI (80 mM) and methylmercury (1 mM) directly suppressed absorption of H 2 15 O by the roots, while methionine sulfoximine (1 mM), abscisic acid (10 pM) and carbonyl cyanide m-chlorophenylhydrazone (10 mM) were transported to the leaves and enhanced stomatal closure, reducing H 2 15 O translocation.

The toxicity of vanadium (V) and the effect of chitosan has been investigated on soybean, rice, wheat and barley. Wheat and barley were sensitive to V than rice and soybean but all seedlings of these plants were damaged at 2.5 μg/ml V (in VCl 3 ). These damages were reduced by application of radiation-degraded chitosan. The recovery of growth and reduction of V levels in seedlings were obtained by the treatments with 10–100 μg/ml chitosan irradiated at 70–200 kGy of γ -rays in 1% solution. The reductions of V and Fe contents in plants were due to the ability of chitosan to form chelate complexes with metals in solution. The result of BAS analysis shows that the absorption and transportation of 48 V to the leaf from root was suppressed with irradiated chitosan. Therefore, it can be concluded that chitosan irradiated at suitable doses (ca. 100 kGy) is effective as plant growth promoters and heavy metal eliminators in crop production.

The ammonium ion is an indispensable nitrogen source for crops, especially paddy rice ( Oryza sativa L. cv Nipponbare). Until now, it has been impossible to measure ammonium uptake and nitrogen movement in plants in real time. Using the new technologies of PETIS (positron emitting tracer imaging system) and PMPS (positron multi-probe system), we were able to visualize the real time translocation of nitrogen and water in rice plants. We used positron-emitting 13 N-labeled ammonium ( 13 NH 4 + ) and 15 O-water to monitor the movement. In plants cultured under normal conditions, 13 NH 4 + supplied to roots was taken up, and a 13 N signal was detected at the discrimination center, the basal part of the shoots, within 2 minutes. This rapid translocation of 13 N was almost completely inhibited by a glutamine synthetase inhibitor, methionine sulfoximine. In general, nitrogen deficiency enhanced 13 N translocation to the discrimination center. In the dark, 13 N translocation to the discrimination center was suppressed to 40% of control levels, whereas 15 O-water flow from the root to the discrimination center stopped completely in the dark. In abscisic acid-treated rice, 13 N translocation to the discrimination center was doubled, whereas translocation to leaves decreased to 40% of control levels. Pretreatment with NO 3 − for 36 hours increased 13 N translocation from the roots to the discrimination center to 5 times of control levels. These results suggest that ammonium assimilation (from the roots to the discrimination center) depends passively on water flow, but actively on NH 4 + -transporter(s) or glutamine synthetase(s).

We present real time vanadate (V5+ ) uptake imaging in acowpea plant by positron emitting tracer imaging system (PETIS). Vanadium-48was produced by bombarding a Sc foil target with 50 MeV α -particlesat Takasaki Ion Accelerators for Advanced Radiation Application (TIARA) AVFcyclotron. Then 48 V was added to the culture solution to investigatethe V distribution in a cowpea plant. The real time uptake of the 48V was monitored by PETIS. We measured the distribution of 48Vin a whole plant after 3, 6 and 20 hours of V treatment by Bio-imaging AnalyzerSystem (BAS). After the 20 hour treatment, vanadate was detected at the up-groundpart of the plant. To know the effect of V uptake on plant activity, 18F-labeled water uptake was analyzed by PETIS. When a cowpea plantwas treated with V for 20 hours before 18 F-labeled water uptakeexperiment, the total amount of 18F-labeled water absorption wasdrastically decreased. Results suggest the inhibition of water uptake wasmainly caused by the vanadate already moved to the up-ground part of the plant.

15P-water flow from the roots to the top in tomato (Lycopersicon esculentum Mill.) and rice (Oryza sativa L.) plants was visualized with time using a positron-emitting tracer imaging system (PETIS). The 15O-water flow was switched on by light and completely stopped in the dark. The flow rate in the stem of tomato and the shoot of rice at a light intensity of 500 μmol·m−2·s−1 was 1.9 and 0.4 cm min−1, respectively.

The positron emitting tracer-imaging system (PETIS) was used to determine whether it was possible to obtain on image of 13N-distribution in common bean in which a single root was fed with a liquid medium containing nitrate at different concentrations with different 13N specific activities. The distribution of the images of the 13N atoms in the root could be obtained over a wide range of nitrate concentrations and 13N specific activities in the medium. As for nitrate stress on leguminous root nodulation, the positional relationship between the nitrate uptake sites and root hair just elongating area, where rhizobia capably initiate their infection, was studied in common bean and soybean. PETIS gave direct evidence that single roots of both common bean and soybean showed one or two dense 13N-distribution areas after 2 min pulse-feeding of 13N03 -. These areas remained stable over 30 min, and the first dense site, which was common in all the examined roots, extended over ca. 1 cm above the root apex....

The lelvels of seven heavy metals and their toxicity towardGanoderma lucidum under various cultivation conditions were assessed. The contents of Mn, Cu, Zn, Cd, Hg, Pb and U in the fruitbodies of cultivatedG. lucidum, and sawdust substrates were determined to be at trace levels for U, 0.01–0.1 μg/g for Cd and Hg, and 1–5 μg/g for Pb, 10–120 μg/g for Mn, Cu and Zn. The effects of heavy metals, on the growth of mycelia ofG. lucidium in pure cultures were examined over a wide range of concentrations (10–3,000 μg/ml), and their toxicities were found to decrease in the order: Hg>Cd>Cu>U>Pb>Mn=Zn. The translocation and accumulation of Zn from contaminated substrates (at 10 μg/g) in fruitbodies were investigated by using65Zn tracer, andG. lucidum was found to take up Zn with an efficiency of >60%, leading to accumulation of >100 μ/g, in fruitbodies and >80 μ/g Zn in basidiospores.

The radiation effects on the carbohydrates in chicken ovomucoid, a protease inhibitor as a typical allergenic glycoprotein of egg white, were investigated to clarify its role for the trypsin inhibitory activity on irradiation. The trypsin inhibitory activity of chicken ovomucoid decreased exponentially as a function of the radiation dose. In O 2 , the inactivation of chicken ovomucoid was protected remarkably in comparison to that in N 2 . With protein blotting, protein was degraded in O 2 and aggregated in N 2 . The patterns of carbohydrate blotting were similar to those of protein blotting. These results show that there could be a structural interrelationship between the active site and carbohydrate moiety. Sugar chains in a low molecular weight fraction (MW γ -radiation and those in O 2 were higher than in N 2 . From the high-performance liquid chromatography (HPLC) patterns of the degradation of sugar chains, all peaks of the oligosaccharides decreased with an increase of radiation dose and more remarkable in O 2 than in N 2 . It shows that the oligosaccharides of ovomucoids could be released sighificantly in O 2 by the degradation associated with γ -radiation. These results suggest that oxygen could play a protective role in the inactivation of ovomucoids by the removal of reducing species generated by γ -radiation and ionizing radiation could cause overall conformational changes by the degradation and release of oligosaccharides, as well as alter the bioactivity of ovomucoid.

Responses of mycelia ofGanoderma lucidum to vanadium, selenium and germanium were examined over a wide range of concentrations (10–1, 120 μg/ml) in pure culture. Se and V were found to be highly toxic, but Ge was not toxic at the levels tested.Ganododerma lucidum cultivated on substrates of sawdust with V (30–80 μg/g) developed mature fruitbodies, but the bioaccumulation of V was quite low (2.5–7 μg/g in pileus, 12.5–21.5 μg/g in stipe and

The positron emitters11C,13N and18F and their labeled compounds have been produced for studies on plants using a newly developed positron emitting tracer imaging system. Although this system covers, at present, a limited area in a plant, the distribution of the positron emitter fed into the plant can be visualized dynamically. Further development of positron-emitter-labeled compounds is expected to elucidate the physiological function of plants in vivo.

The upgrading of sugar cane bagasse with natural rubber waste by irradiation and three mushroom fungi was studied in order to produce economical animal feed and mushrooms. The protein concentrations changed a little at 0, 1% of NRSP but greatly increased at 5% NRSP concentration with irradiation and nonirradiation in liquid fermentation. The filtration rate decreased at 5% NRSP concentration after liquid fermentation but it increased by 2.7, 10.2, 11.1 times with irradiation for P. sajor-caju , C. phlytidosprorus and P. flavellatus . The amounts of crude fiber were decreased due to the irradiation and the different concentration of NRSP after 42 days of solid fermentation by P. sajor-caju .

Endohedral 133Xe-higher fullerenes (133Xe@C76 and 133Xe@C84) were produced by implantation of 133Xe ions using an isotope separator. A high performance liquid chromatography (HPLC) analysis showed that the peak of endohedral 133Xe-higher fullerenes shifted backward from that of empty fullerenes, suggesting a possibility of the separation of endohedral 133Xe-higher fullerenes from empty fullerenes. The yields of endohedral 133Xe-fullerenes were in the order of 133Xe@C76

The transport of a positron-emitting isotope introduced into a plant was dynamically followed by a special observation apparatus called ‘Positron-Emitting Tracer Imaging System’ to observe the damage and recovery functions of plants in vivo. In the system, annihilation γ-rays from the positron emitter are detected with two planar detectors (5 × 6 cm2). The water containing ca. 5 MBq/ml of 18F was fed to the cut stem of soybean for 2 min and then the images of tracer activity were recorded for 30–50 min. When the midrib of a leaf near the petiole was cut just before measurement, the activity in the injured leaf was decreased but detected even at the apex. This result suggests that the damaged leaf recovered the uptake of water through the lamina. Maximum tracer activities in leaves of unirradiated plant were observed within 10 min, whereas those of irradiated plant at 100 Gy were observed after over 25 min. The final activity of irradiated plant after 30 min was lower than that of unirradiated plant. In case of beans, there was a difference in the absorption behavior of the 18F-labeled water between unirradiated and irradiated samples. These results show that the system is effective to observe the uptake and transportation of water containing positron emitting tracer for the study of damage and recovery functions of plants.

Antimicrobial activity of irradiated chitosan was studied against Escherichia coli B/r. Irradiation of chitosan at 100 kGy under dry conditions was effective in increasing the activity, and inhibited the growth of E coli completely. The molecular weight of chitosan significantly decreased with the increase in irradiation dose, whereas the relative surface charge of chitosan was decreased only 3% by 100 kGy irradiation. Antimicrobial activity assay of chitosan fractionated according to molecular weight showed that 1 x 10 5 -3 x 10 5 fraction was most effective in suppressing the growth of E coli. This fraction comprised only 8% of the 100 kGy irradiated chitosan. On the other hand, chitosan whose molecular weight was less than 1 x 10 5 had no activity. The results show that low dose irradiation, specifically 100 kGy, of chitosan gives enough degradation to increase its antimicrobial activity as a result of a change in molecular weight.

Radiation effects on biochemical properties of ovomucoid, a protease inhibitor which is a typical allergenic protein of egg, were investigated. Trypsin inhibiting activity of ovomucoid decreased exponentially and the inactivation was larger in N2 than in O2. The antigenicity of ovomucoid measured by ELISA was decreased exponentially and the inactivation curves were almost the same as that of trypsin inhibiting activity. These results show that the oxygen protects the inactivation and destruction of antigenicity of ovomucoid while it enhances the reaction with ovalbumin. The effect of radiolytically generated radical anions; a pronounced protective action of (CNS)2− and a slight accelerate action of Br2− were the same both on decrease in trypsin inhibiting activity and antigenicity. SDS-PAGE showed that the change in protein structure was also protected by (CNS)2−. These results suggest that the changes by radiation were not mainly due to the destruction of essential amino acid residues but to the conformational change.

The effect of radiation pasteurization of sugar cane bagasse and rice straw and fermentation using various strains of fungi were studied for upgrading of cellulosic wastes. The initial contamination by fungi and aerobic bacteria both in bagasse and straw was high. The doses of 30 kGy for sterilization and 8 kGy for elimination of fungi were required. Irradiation effect showed that rice straw contained comparatively radioresistant microorganisms. It was observed that all the fungi (Hericium erinacium, Pleurotus djamor, Ganoderma lucidum, Auricularia auricula, Lentinus sajor-caju, Coriolus versicolor, Polyporus arcularius, Coprinus cinereus) grow extending over the entire substrates during one month after inoculation in irradiated bagasse and rice straw with 3% rice bran and 65% moisture content incubated at 30°C. Initially, sugar cane bagasse and rice straw substrates contained 39.4% and 25.9% of cellulose, 22.9% and 26.9% of hemicellulose, and 19.6% and 13.9% of lignin + cutin, respectively. Neutral detergent fibre (NDF) values decreased significantly in sugar cane bagasse fermented byG. lucidum, A. auricula andP. arcularius, and in rice straw fermented by all the 8 strains of fungi. Acid detergent fibre (ADF) values also decreased in bagasse and rice straw fermented by all the fungi.P. arcularius, H. erinacium, G. lucidum andC. cinereus were found to be the most effective strains for delignification of sugar cane bagasse.

The production of animal feeds and mushrooms from oil palm cellulosic wasres by radiation and fermentation has been investigated in order to utilize the agro-resources and to reduce the smoke pollution. The process is as follows: decontamination of microorganisms in fermentation media of empty fruit bunch of oil palm (EFB) by irradiation, inoculation of useful fungi, and subsequently production of proteins and edible mushrooms. The dose of 25 kGy was required for the sterilization of contaminating bacteria whereas the dose of 10 kGy was enough to eliminate the fungi. Among many kinds of fungi tested, C. cinereus and P. sajor-caju were selected as the most suitable microorganism for the fermentation of EFB. The protein content of the product increased to 13 % and the crude fiber content decreased to 20% after 30 days of incubation with C. cinereus at 30°C in solid state fermentation. P. sajor-caju was suitable for the mushroom production on EFB with rice bran.

Degradation percentages of pectates by exopolygalacturonate lyase, exopolygalacturonase (exo-PG), and exo-PG and endopolygalactur-onase in combination can be accurately measured by our HPLC method. This enzymic-HPLC method was successfully used for the structural analysis of pectic substances, which are usually composed of two distinct regions of linear galacturonan (GN) and branched rhamnogalacturonan (RG). The GN regions were classified for convenience into four categories and the relative amounts of each GN region were represented by α∼δ, viz., (α) GN chains having a nonreducing end, (β) those having a reducing end, (γ) those having both reducing and nonreducing ends, and no neutral sugar residues, and (δ) those interposed between two RG regions; in addition, (∊) the amount of the residual galacturonate, mainly comprised in the RG regions. The procedure for the determination of α∼∊ was described in detail, and the results of application to several pectate preparations were reported

Salinity stress affects photosynthate partitioning between sources and sinks of plants, but how it affects these systems is less well understood. Because sources and sinks are closely tied, any adverse effect under suboptimal conditions on one of these is often misinterpreted for an effect on the other. Carbon partitioning is indispensable for stress resistance and good plant growth. In the present study, carbon partitioning in tomato plants ( Lycopersicon esculentum L. cv. Momotarou) in a saline (NaCl) environment was studied by feeding radioactive 11 C and stable 13 C isotopes. Pulse-chases were conducted to measure the spatial and temporal distribution of 13 C. 13 C was measured by a standard conventional technique, but 11 C distribution was monitored using a positron-emitting tracer imaging system (PETIS). Salt stress resulted in reduced carbon translocation toward roots. The majority of the photosynthate accumulated in the leaf. We also observed that the reduction in translocation of carbon occurred well before the salt stress symptoms of reduced photosynthesis and reduced plant growth in salt-exposed plants. The effect on sink activity was also shown by a decrease in stem diameter. In addition, PETIS analysis of 11 C translocation indicated that carbon translocation to roots was inhibited under salt conditions without a direct effect on leaf Na accumulation or osmotic stress. These results suggest that NaCl has direct effects on plants, inhibiting carbon partitioning within a few hours of salt exposure without inhibition of source activity.

Position emitting tracer imaging system (PETIS) and carbon-11-labeled carbon dioxide (11CO2) can image photosynthesis process in plant leaf, and 11C kinetics make it possible to estimate physiological function parameters in photosynthesis. With an exposure 11CO2-gas to a leaf, PETIS experiments were performed under four light conditions on a single leaf. In order to estimate the rate constants of photosynthesis parameters, time activity curves of 11CO2 gas input and leaf response were fitted to an appropriate compartmental tracer kinetic model, which applies influx and efflux for photo-assimilation and photosynthate delivery rate constants respectively. These results are reasonable response against light environment and important to discuss photosynthesis in plant physiology and agriculture

A radioisotope-production facility at the Takasaki Ion Accelerators for Advanced Radiation Application (TIARA) of the Japan Atomic Energy Agency (JAEA) allows us to produce radioisotopes using accelerated beam from an AVF cyclotron. The facility has been constructed for research and development of potentially useful radioisotopes in medical research and physiological studies of plants. Experiments in these fields require a variety of radioisotopes and their labeled compounds during a limited machine time. Laboratory equipment has been designed and installed to treat high-level radioactivity and to irradiate a variety of targets (solid, liquid, gas....). The irradiation apparatus consists of two solid target chambers and a compact revolver equipped with six target chambers for gas and liquid targets, set on the movable table. It is possible to exchange rapidly and remotely the target chambers. The solid-target transfer system realized to carry automatically the irradiated target from irradiation position to the shielded-cell by a truck. Use of this facility enables us to produce routinely positron-emitting tracers for plant physiology and to develop the production method of carrier-free radioisotopes for medical use

Summary 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.

The positron emitting tracer imaging system (PETIS) and carbon-11-labeled carbon dioxide (11CO2) can image carbon movement during photosynthesis in a plant leaf, and 11C kinetics make it possible to estimate physiological function parameters in those photosynthesis processes. With an exposure 11CO2 gas to a leaf, PETIS experiments were performed iteratively under dark and light environments on a single leaf. In order to estimate the rate constants of photosynthetic parameters, time activity curves of 11CO2 gas input and leaf response were fitted to an appropriate compartmental tracer kinetic model, which applies influx and efflux for photo-assimilation and sucrose export rate constants respectively. Results obtained from the kinetic analysis are consistent with physiological knowledge and important to discuss photosynthesis in plant physiology and agriculture. In addition, the proposed method in this paper produce parametric images of photosynthetic functions on a pixel-by-pixel basis successfully, in other words, molecular imaging for plant study is demonstrated.

The Positron-Emitting Tracer Imaging System (PETIS) is introduced for monitoring the distribution of (11)C-labelled photoassimilates in Sorghum. The obtained two-dimensional image data were quantitatively analysed using a transfer function analysis approach. While one half of a Sorghum root in a split root system was treated with either 0, 100, or 500 mM NaCl dissolved in the nutrient solution, tracer images of the root halves and the lower stem section were recorded using PETIS. From the observed tracer levels, parameters were estimated, from which the mean speed of tracer transport and the proportion of tracer moved between specified image positions were deduced. Transport speed varied between 0.7 and 1.8 cm min(-1) with the difference depending on which part of the stem was involved. When data were collected in the lowest 0.5-1 cm of the stem, which included the point where the roots emerge, transport speed was less. Rapid changes in NaCl concentration, from 0 to 100 mM, resulted in short-term increases of assimilate import into the treated root. This response represented a transient osmotic effect, that was compensated for in the medium-term by osmotic adaptation. Higher concentrations of NaCl (500 mM) resulted in distinctly less photoassimilate transport into the treated root half. The present results agree with earlier observations, showing that transport of (11)C-labelled photoassimilates measured with the PETIS detector system can be quantified using the method of input-output analysis. It is worth noting that with the PETIS detector system, areas of interest do not need to be defined until after data collection. This means that unexpected behaviour of a plant organ will be seen, which is not necessarily the case with conventional detector systems looking at predefined areas of interest.

Effect of γ-irradiation and combination treatment with alkali on oil palm empty fruit bunch (EFB) was studied to enhance the efficiency of enzymatic digestibility. γ-Irradiation enhanced the enzymatic digestibility, and the amounts of released galacturonic acid and neutral sugars from EFB by Driselase were increased with an increase in irradiation dose. The main components of neutral sugars released from EFB were glucose and xylose. EFB treated with alkali and 50 kGy irradiation released 1200 μg g -1 of xylose that was twice amount of glucose showing that the combination treatment was effective especially to increase the digestibility of xylan.

The upper part of a nodulated soybean root hydroponically cultured in a glass bottle was monitored using a computer microscope under controlled environmental conditions, and the diameter of individual nodules was measured from 10-24 d after planting. The diameter of a root nodule attached to the primary root increased from 1 mm to 6 mm for 2 weeks under nitrogen-free conditions. The increase in diameter of the nodules was almost completely stopped after 1 d of supplying 5 mM nitrate, and was due to the cessation of nodule cell expansion. However, nodule growth quickly returned to the normal growth rate following withdrawal of nitrate from the solution. The reversible depression of nodule growth by nitrate was similar to the restriction of photoassimilate supply by continuous dark treatment for 2 d followed by normal light/dark conditions. In addition, the inhibitory effect of nitrate was partially alleviated by the addition of 3% (w/v) sucrose to the medium. Plant leaves were exposed to (11)C or (14)C-labelled carbon dioxide to investigate the effects of 5 mM nitrate on the translocation and distribution of photosynthates to nodules and roots. Supplying 5 mM nitrate stimulated the translocation rate and the distribution of labelled C in nitrate-fed parts of the roots. However, the (14)C partitioning to nodules decreased from 9% to 4% of total (14)C under conditions of 5 mM nitrate supply. These results indicate that the decrease in photoassimilate supply to nodules may be involved in the quick and reversible nitrate inhibition of soybean nodule growth.

Quantitative analysis of the two-dimensional image data obtained with the positron-emitting tracer imaging system (PETIS) for plant physiology has been carried out using a transfer function analysis method. While a cut leaf base of Chinese chive (Allium tuberosum Rottler) or a cut stem of soybean (Glycine max L.) was immersed in an aqueous solution containing the [18F] F− ion or [13N]NO3− ion, tracer images of the leaf of Chinese chive and the trifoliate of soybean were recorded with PETIS. From the time sequence of images, the tracer transfer function was estimated from which the speed of tracer transport and the fraction moved between specified image positions were deduced.

[11C]Methionine was supplied through barley roots and the 11C signal was followed for 90 min using a real-time imaging system (PETIS), with subsequent development of autoradiographic images of the whole plant. In all cases, [11C]methionine was first translocated to the 'discrimination center', the basal part of the shoot, and this part was most strongly labeled. Methionine absorbed by the roots of the plants was subsequently translocated to other parts of the plant. In Fe-deficient barley plants, a drastic reduction in [11C]methionine translocation from the roots to the shoot was observed, while a greater amount of 11C was found in the leaves of Fe-sufficient or methionine-pretreated Fe-deficient plants. Treatment of Fe-deficient plants with aminooxyacetic acid, an inhibitor of nicotianamine aminotransferase, increased the translocation of [11C]methionine to the shoot. The retention of exogenously supplied [11C]methionine in the roots of Fe-deficient barley indicates that the methionine is used in the biosynthesis of mugineic acid phytosiderophores in barley roots. This and the absence of methionine movement from shoots to the roots suggest that the mugineic acid precursor methionine originates in the roots of plants.

A simple and reliable method for the determination of NH4+, K+, Na+, aspartic acid, asparagine, glutamine, and alanine by ion chromatography has been developed. It is suitable for monitoring changes of nitrogen metabolism in soybean because it can accurately measure concentrations o asparagine and NH4+, two key substances for nitrogen storage and transport in this plant species Analysis of asparagine distribution in soybean indicated that higher levels (up to 18.4 micromol g(-1) of fresh mass) occur in stems and lower levels in roots (2.0 micromol g(-1) of fresh mass) and leaves (1.6 micromol g(-1) of fresh mass). When the herbicide metsulfuron-methyl (0.5, 5, and 50 ppb) was applied via the nutrient solution to the root system, asparagine concentrations increased 3-6 times in stems roots, and leaves. Metsulfuron-methyl is known to impair the synthesis of branched amino acids and, in consequence, protein synthesis. Thus, nitrogen consumption was limited, leading to ar accumulation of asparagine. The possible use of this physiological response in agricultural practice to identify herbicide stress in soybean and to detect low-level residues of sulfonylurea herbicides ir the soil is discussed.

Non-nodulated soybean (Glycine max (L.) Merr.) plants were cultivated hydroponically under N-sufficient (5 mM NaNO 3 ) or N-deficient (0.5 mM NaNO 3 ) conditions. 13 N- or 15 N- labelled nitrate was fed to the cut end of the stems, and the accumulation of nitrate-derived N in the pods, nodes and stems was compared. Real-time images of 13 N distribution in stems, petioles and pods were obtained using a Positron Emitting Tracer Imaging System for a period of 40 min. The results indicated that the radioactivity in the pods of N-deficient plants was about 10 times higher than that of N-sufficient plants, although radioactivity in the stems and nodes of N-deficient versus N-sufficient plants was not different. A similar result was obtained by supplying 15NO3- to cut soybean shoots for 1 h. The fact that the N translocation into the pods from NO 3 - fed to the stem base was much faster in N-deficient plants may be due to the strong sink activity of the pods in N-deficient plants. Alternatively, the redistribution of N from the leaves to the pods via the phloem may be accelerated in N-deficient plants. The temporal accumulation of 13 NO 3 in nodes was suggested in both N-sufficient and N-deficient plants. In one 13 NO 3 - pulse-chase experiment, radioactivity in the stem declined rapidly after transferring the shoot from the 13 NO 3 - solution to non-labelled NO 3 ; in contrast, the radioactivity in the node declined minimally during the same time period.

Nitrogen movement through the xylem vessels and sieve tubes in rice plants was studied using xylem and phloem sap analysis in combination with stable and radioactive nitrogen isotope techniques.

The transport of l8F-water introduced into irradiated plants was followed dynamically by the detection of annihilation γ -rays from a positron emitter using a “Positron-Emitting Tracer Imaging System”. Tracer activities at the vein of top leaf of unirradiated soybean (Glysine max) were observed in a short time, whereas those in plants irradiated with a doses up to 100 Gy were delayed with increase in dose. In the case of rose (Rosa sp. Rote Rose), radiation using a dose of 800 Gy for disinfestation as a quarantine procedure inhibited flowering. Uptake and transport of 18F-water through the cut stem was changed significantly by irradiation. These results show that the system is effective in observing the damage of irradiated plants and is useful for the study of physiological functions of plants.