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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 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.

To know the effect of drought stress on two cultivars of cowpea, drought tolerant (DT) and drought sensitive (DS), and to estimate vanadium treatment on plant activity, we performed real time 18F labeled water uptake measurement by PETIS. Fluoride-18 was produced by bombarding a cubic ice target with 50 MeV protones using TIARA AVF cyclotron. Then 18F labeled water was applied to investigate water movement in a cowpea plant. Real time water uptake manner could be monitored by PETIS. After the analysis by PETIS, we also measured the distribution of 18F in a whole plant by BAS. When a cowpea plant was treated with drought stress, there was a difference in water uptake manner between DT and DS cultivar. When a cowpea plant was treated with V for 20 hours before the water uptake experiment, the total amount of 18F labeled water absorption was found to be drastically decreased.

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 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 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.

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.

[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.

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.

[ 11 C]methionine was supplied to Fe-deficient and Fe-sufficient barley plants through a single leaf, and real time 11 C movement was monitored using a Positron Emitting Tracer Imaging System (PETIS). In Fe-deficient plants, [ 11 C]methionine was translocated from the tip of the absorbing leaf to the 'discrimination centre' located at the base of the shoot, and then retranslocated to all the chlorotic leaves within 60 min, while a negligible amount was retranslocated to the roots. In Fe-sufficient plants, methionine was translocated to the discrimination centre and then only to the newest leaf on the main shoot within 60 min. A negligible amount was also retranslocated to the roots. In conclusion, methionine from the above-ground parts of a plant is not a precursor of mugineic acid under Fe-deficiency. The discrimination centre is suggested to play a vital role in the distribution of mineral elements and metabolites in gramin-aceous monocots.