Naoki Kawachi

Tracing Cadmium from Culture to Spikelet: Noninvasive Imaging and Quantitative Characterization of Absorption, Transport, and Accumulation of Cadmium in an Intact Rice Plant

We characterized the absorption and short-term translocation of cadmium (Cd) in rice (Oryza sativa ‘Nipponbare’) quantitatively using serial images observed with a positron-emitting tracer imaging system. We fed a positron-emitting 107Cd (half-life of 6.5 h) tracer to the hydroponic culture solution and noninvasively obtained serial images of Cd distribution in intact rice plants at the vegetative stage and at the grain-filling stage every 4 min for 36 h. The rates of absorption of Cd by the root were proportional to Cd concentrations in the culture solution within the tested range of 0.05 to 100 nm. It was estimated that the radial transport from the culture to the xylem in the root tissue was completed in less than 10 min. Cd moved up through the shoot organs with velocities of a few centimeters per hour at both stages, which was obviously slower than the bulk flow in the xylem. Finally, Cd arrived at the panicles 7 h after feeding and accumulated there constantly, although no Cd was observed in the leaf blades within the initial 36 h. The nodes exhibited the most intensive Cd accumulation in the shoot at both stages, and Cd transport from the basal nodes to crown root tips was observed at the vegetative stage. We conclude that the nodes are the central organ where xylem-to-phloem transfer takes place and play a pivotal role in the half-day travel of Cd from the soil to the grains at the grain-filling stage.

Rapid Quantitative Measurement of CMRO2 and CBF by Dual Administration of 15O-Labeled Oxygen and Water During a Single PET Scan—a Validation Study and Error Analysis in Anesthetized Monkeys

Cerebral blood flow (CBF) and rate of oxygen metabolism (CMRO2) may be quantified using positron emission tomography (PET) with 15O-tracers, but the conventional three-step technique requires a relatively long study period, attributed to the need for separate acquisition for each of 15O2, H215O, and C15O tracers, which makes the multiple measurements at different physiologic conditions difficult. In this study, we present a novel, faster technique that provides a pixel-by-pixel calculation of CBF and CMRO2 from a single PET acquisition with a sequential administration of 15O2 and H215O. Experiments were performed on six anesthetized monkeys to validate this technique. The global CBF, oxygen extraction fraction (OEF), and CMRO2 obtained by the present technique at rest were not significantly different from those obtained with three-step method. The global OEF (gOEF) also agreed with that determined by simultaneous arterio-sinus blood sampling (gOEFA–V) for a physiologically wide range when changing the arterial PaCO2 (gOEF = 1.03gOEFA–V +0.01, P<0.001). The regional values, as well as the image quality were identical between the present technique and three-step method for CBF, OEF, and CMRO2. In addition, a simulation study showed that error sensitivity of the present technique to delay or dispersion of the input function, and the error in the partition coefficient was equivalent to that observed for three-step method. Error sensitivity to cerebral blood volume (CBV) was also identical to that in the three-step and reasonably small, suggesting that a single CBV assessment is sufficient for repeated measures of CBF/CMRO2. These results show that this fast technique has an ability for accurate assessment of CBF/CMRO2 and also allows multiple assessment at different physiologic conditions.

Real-time imaging and analysis of differences in cadmium dynamics in rice cultivars (Oryza sativa) using positron-emitting107Cd tracer

BackgroundRice is a major source of dietary intake of cadmium (Cd) for populations that consume rice as a staple food. Understanding how Cd is transported into grains through the whole plant body is necessary for reducing rice Cd concentrations to the lowest levels possible, to reduce the associated health risks. In this study, we have visualized and quantitatively analysed the real-time Cd dynamics from roots to grains in typical rice cultivars that differed in grain Cd concentrations. We used positron-emitting107Cd tracer and an innovative imaging technique, the positron-emitting tracer imaging system (PETIS). In particular, a new method for direct and real-time visualization of the Cd uptake by the roots in the culture was first realized in this work.ResultsImaging and quantitative analyses revealed the different patterns in time-varying curves of Cd amounts in the roots of rice cultivars tested. Three low-Cd accumulating cultivars (japonica type) showed rapid saturation curves, whereas three high-Cd accumulating cultivars (indica type) were characterized by curves with a peak within 30 min after107Cd supplementation, and a subsequent steep decrease resulting in maintenance of lower Cd concentrations in their roots. This difference in Cd dynamics may be attributable to OsHMA3 transporter protein, which was recently shown to be involved in Cd storage in root vacuoles and not functional in the high-Cd accumulating cultivars. Moreover, the PETIS analyses revealed that the high-Cd accumulating cultivars were characterized by rapid and abundant Cd transfer to the shoots from the roots, a faster transport velocity of Cd to the panicles, and Cd accumulation at high levels in their panicles, passing through the nodal portions of the stems where the highest Cd intensities were observed.ConclusionsThis is the first successful visualization and quantification of the differences in whole-body Cd transport from the roots to the grains of intact plants within rice cultivars that differ in grain Cd concentrations, by using PETIS, a real-time imaging method.

Experimental Results of the Gamma-Ray Imaging Capability With a Si/CdTe Semiconductor Compton Camera

A semiconductor Compton camera that combines silicon (Si) and cadmium telluride (CdTe) detectors was developed, and its imaging capability was examined with various kinds of gamma-ray targets such as a point source, arranged point sources and an extended source. The camera consists of one double-sided Si strip detector and four layers of CdTe pad detectors, and was designed to minimize the distance between a scatterer and the target. This is because the spatial resolution with Compton imaging improves as the target approaches the scatterer. This new camera realizes a minimum distance of 25 mm. By placing the target at a distance of 30 mm from the detector, resolving power better than 3 mm was demonstrated experimentally for a 364 keV (131I) gamma-ray. Positional determination with accuracy of 1 mm was also demonstrated. As a deconvolution method, we selected the iteration algorithm (called List-Mode Expectation-Maximizing Maximum Likelihood), and applied it to several kinds of experimental data. The Compton back projection images of the arranged point sources and an extended object were successfully deconvolved.

Kinetic Analysis of Carbon-11-Labeled Carbon Dioxide for Studying Photosynthesis in a Leaf Using Positron Emitting Tracer Imaging System

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

Application of glutathione to roots selectively inhibits cadmium transport from roots to shoots in oilseed rape

Glutathione is a tripeptide involved in various aspects of plant metabolism. This study investigated the effects of the reduced form of glutathione (GSH) applied to specific organs (source leaves, sink leaves, and roots) on cadmium (Cd) distribution and behaviour in the roots of oilseed rape plants (Brassica napus) cultured hydroponically. The translocation ratio of Cd from roots to shoots was significantly lower in plants that had root treatment of GSH than in control plants. GSH applied to roots reduced the Cd concentration in the symplast sap of root cells and inhibited root-to-shoot Cd translocation via xylem vessels significantly. GSH applied to roots also activated Cd efflux from root cells to the hydroponic solution. Inhibition of root-to-shoot translocation of Cd was visualized, and the activation of Cd efflux from root cells was also shown by using a positron-emitting tracer imaging system (PETIS). This study investigated a similar inhibitory effect on root-to-shoot translocation of Cd by the oxidized form of glutathione, GSSG. Inhibition of Cd accumulation by GSH was abolished by a low-temperature treatment. Root cells of plants exposed to GSH in the root zone had less Cd available for xylem loading by actively excluding Cd from the roots. Consequently, root-to-shoot translocation of Cd was suppressed and Cd accumulation in the shoot decreased.

From laboratory to field: OsNRAMP5-knockdown rice is a promising candidate for Cd phytoremediation in paddy fields.

Previously, we reported that OsNRAMP5 functions as a manganese, iron, and cadmium (Cd) transporter. The shoot Cd content in OsNRAMP5 RNAi plants was higher than that in wild-type (WT) plants, whereas the total Cd content (roots plus shoots) was lower. For efficient Cd phytoremediation, we produced OsNRAMP5 RNAi plants using the natural high Cd-accumulating cultivar Anjana Dhan (A5i). Using a positron-emitting tracer imaging system, we assessed the time-course of Cd absorption and accumulation in A5i plants. Enhanced 107Cd translocation from the roots to the shoots was observed in A5i plants. To evaluate the phytoremediation capability of A5i plants, we performed a field experiment in a Cd-contaminated paddy field. The biomass of the A5i plants was unchanged by the suppression of OsNRAMP5 expression; the A5i plants accumulated twice as much Cd in their shoots as WT plants. Thus, A5i plants could be used for rapid Cd extraction and the efficient phytoremediation of Cd from paddy fields, leading to safer food production.

Real-time imaging of nitrogen fixation in an intact soybean plant with nodules using 13N-labeled nitrogen gas

Abstract Real-time images of nitrogen fixation in an intact nodule of hydroponically cultured soybean (Glycine max [L] Merr.) were obtained. In the present study, we developed a rapid method to produce and purify 13N-labeled radioactive nitrogen gas (half life: 9.97 min). 13N was produced from a 16O (p, α) 13N nuclear reaction. The target chamber was filled with CO2 and irradiated for 10 min with protons at an energy of 18.3 MeV and an electric current of 5 μA, which was delivered from a cyclotron. All CO2 in the collected gas was absorbed and removed with powdered soda-lime in a syringe and replaced with helium gas. The resulting gas was injected into gas chromatography and separated and a 35 mL fraction, including the peak of [13N]-nitrogen gas, was collected by monitoring the chromatogram. The obtained gas was mixed with 10 mL of O2 and 5 mL of N2 and used in the tracer experiment. The tracer gas was fed into the underground part of intact nodulated soybean plants and serial images of the distribution of 13N were obtained non-invasively using a positron-emitting tracer imaging system (PETIS). The rates of nitrogen fixation of the six test plants were estimated to be 0.17 ± 0.10 μmol N2 h−1 from the PETIS image data. The decreasing rates of assimilated nitrogen were also estimated to be 0.012 ± 0.011 μmol N2 h−1. In conclusion, we successfully observed nitrogen fixation in soybean plants with nodules non-invasively and quantitatively using [13N]N2 and PETIS.

Analysis of NO3 interception of the parasitic angiosperm Orobanche spp. using a positron-emitting tracer imaging system and 13NO3−: A new method for the visualization and quantitative analysis of the NO3 interception ratio

Abstract The root parasitic plants Orobanche spp. (broomrapes) seriously affect agricultural production. A visualization and quantitative analytical method for the interception of nutrients was established using a positron-emitting tracer imaging system and 13NO− 3. By using this analytical method that involves volume normalization with 18F− images, the nitrogen nutrient interception ratio of the Orobanche spp. was calculated to be 73.6 ± 3.9% in a host–parasite system of red clover (Trifolium pratense L.).

Kinetic Analysis of Zinc/Cadmium Reciprocal Competitions Suggests a Possible Zn-Insensitive Pathway for Root-to-Shoot Cadmium Translocation in Rice.

BackgroundAmong cereals, rice has a genetic propensity to accumulate high levels of cadmium (Cd) in grains. Xylem-mediated root-to-shoot translocation rather than root uptake has been suggested as the main physiological factor accounting for the genotypic variation observed in Cd accumulation in shoots and grains. Several evidence indicate OsHMA2 – a putative zinc (Zn) transporter – as the main candidate protein that could be involved in mediating Cd- and Zn-xylem loading in rice. However, the specific interactions between Zn and Cd in rice often appear anomalous if compared to those observed in other staple crops, suggesting that root-to-shoot Cd translocation process could be more complex than previously thought. In this study we performed a complete set of competition experiments with Zn and Cd in order to analyze their possible interactions and reciprocal effects at the root-to-shoot translocation level.ResultsThe competition analysis revealed the lack of a full reciprocity when considering the effect of Cd on Zn accumulation, and vice versa, since the accumulation of Zn in the shoots was progressively inhibited by Cd increases, whereas that of Cd was only partially impaired by Zn. Such behaviors were probably dependent on Cd-xylem loading mechanisms, as suggested by: i) the analysis of Zn and Cd content in the xylem sap performed in relation to the concentration of the two metals in the mobile fractions of the roots; ii) the analysis of the systemic movement of 107Cd in short term experiments performed using a positron-emitting tracer imaging system (PETIS).ConclusionsOur results suggest that at least two pathways may mediate root-to-shoot Cd translocation in rice. The former could involve OsHMA2 as Zn2+/Cd2+ xylem loader, whereas the latter appears to involve a Zn-insensitive system that still needs to be identified.