Real-time Radioisotope Imaging Research Articles

Visualization of phosphorus re‐translocation and phosphate transporter expression profiles in a shortened annual cycle system of poplar

Phosphorus (P) is an essential macronutrient for plant growth. In deciduous trees, P is remobilized from senescing leaves and stored in perennial tissues during winter for further growth. Annual internal recycling and accumulation of P are considered an important strategy to support the vigorous growth of trees. However, the pathways of seasonal re-translocation of P and the molecular mechanisms of this transport have not been clarified. Here we show the seasonal P re-translocation route visualized using real-time radioisotope imaging and the macro- and micro-autoradiography. We analysed the seasonal re-translocation P in poplar (Populus alba. L) cultivated under 'a shortened annual cycle system', which mimicked seasonal phenology in a laboratory. From growing to senescing season, sink tissues of 32 P and/or 33 P shifted from young leaves and the apex to the lower stem and roots. The radioisotope P re-translocated from a leaf was stored in phloem and xylem parenchyma cells and redistributed to new shoots after dormancy. Seasonal expression profile of phosphate transporters (PHT1, PHT5 and PHO1 family) was obtained in the same system. Our results reveal the seasonal P re-translocation routes at the organ and tissue levels and provide a foothold for elucidating its molecular mechanisms.

Visualization of 14CO2 gas fixation by plants

Using the real-time radioisotope imaging system (RRIS), we present the carbon dioxide gas fixation process of a soybean plant applying the 14C-labeled gas. When 14CO2 gas was supplied to the selected mature leaf, the fixed carbon, photosynthate, was transferred and accumulated to the younger leaves preferentially within 24 h. When 14CO2 gas was supplied to the younger leaves, fixed carbon was hardly moved. In the case of the pods, fixed 14CO2 gas in the leaf was preferentially transferred to the closest pod.

Evaluation of plastic scintillators for live imaging of 14C-labeled photosynthate movement in plants

We updated real-time radioisotope imaging system (RRIS) to study photosynthate movement in a large-size plant. Earlier, we used a CsI (Tl) scintillator attached to a fiber optic plate (FOS) to convert radiation into light. However, the scintillator size was fixed as 10 cm × 10 cm, which was too small to observe the entire plant. Moreover, the high cost of FOS makes it hard for other researchers to introduce the system in their facilities. Here, we investigated six types of low-priced plastic scintillators, and demonstrated the movement of 14C-labeled photosynthates in a living plant using the most efficient scintillator instead of FOS.

Visualization of how light changes affect ion movement in rice plants using a real-time radioisotope imaging system

We developed a real-time radioisotope imaging system (RRIS) technique that can nondestructively visualize the element absorption and transport process in plants, using not only positron emitters but also commercially available radioisotopes. In this study, we applied RRIS to analyze light effects on ion movement in rice plants. As tracers, 28Mg, 32P, and 45Ca were used. During the first 5 h, dark/light cycle of 3/7 min were set up; the RRIS needs dark conditions during capturing. When the light was ceased 5 h after supplying each tracer, 32P transport from root to shoot decreased immediately. In contrast, 28Mg and 45Ca transport did not change with light conditions. These results suggest that the P transport is dependent on water flow, whereas Mg and Ca transport are independent of water flow.

Visualization of Uptake of Mineral Elements and the Dynamics of Photosynthates in Arabidopsis by a Newly Developed Real-Time Radioisotope Imaging System (RRIS).

Minerals and photosynthates are essential for many plant processes, but their imaging in live plants is difficult. We have developed a method for their live imaging in Arabidopsis using a real-time radioisotope imaging system. When each radioisotope, 22Na, 28Mg, 32P-phosphate, 35S-sulfate, 42K, 45Ca, 54Mn and 137Cs, was employed as an ion tracer, ion movement from root to shoot over 24 h was clearly observed. The movements of 22Na, 42K, 32P, 35S and 137Cs were fast so that they spread to the tip of stems. In contrast, high accumulation of 28Mg, 45Ca and 54Mn was found in the basal part of the main stem. Based on this time-course analysis, the velocity of ion movement in the main stem was calculated, and found to be fastest for S and K among the ions we tested in this study. Furthermore, application of a heat-girdling treatment allowed determination of individual ion movement via xylem flow alone, excluding phloem flow, within the main stem of 43-day-old Arabidopsis inflorescences. We also successfully developed a new system for visualizing photosynthates using labeled carbon dioxide, 14CO2. Using this system, the switching of source/sink organs and phloem flow direction could be monitored in parts of whole shoots and over time. In roots, 14C photosynthates accumulated intensively in the growing root tip area, 200–800 µm behind the meristem. These results show that this real-time radioisotope imaging system allows visualization of many nuclides over a long time-course and thus constitutes a powerful tool for the analysis of various physiological phenomena.

Evaluation of in vivo detection properties of 22Na, 65Zn, 86Rb, 109Cd and 137Cs in plant tissues using real-time radioisotope imaging system

In plant research, radioisotope imaging provides useful information about physiological activities in various tissues and elemental transport between plant organs. To expand the usage of imaging techniques, a new system was developed to visualize beta particles, x-rays and gamma-rays emitted from plant bodies. This real-time radioisotope imaging system (RRIS) visualizes radioactivity after conversion into light with a CsI(Tl) scintillator plate. Herein, the RRIS detection properties of the gamma-ray emitters 22Na, 65Zn, 86Rb, 109Cd and 137Cs were evaluated in comparison with those of radioluminography (RLG) using an imaging plate. The lower quantitative detection limit (Bq mm−2) during a 15 min period ranged from 0.1 to 4, depending on the nuclide, similar to that of RLG. When the quantitative ability to detect radiation from various Arabidopsis tissues was analyzed, the quantitative capability in silique and the thick internode tended to be low. In an EGS5 simulation, beta particles were the greatest contributors to RRIS imaging of 22Na, 86Rb and 137Cs, and low-energy x-rays contributed significantly to 65Zn and 109Cd detection. Thus, both self-absorption and air space between the sample and scintillator surface could impair quantitative RRIS imaging. Despite these issues, RRIS is suggested for quantitative time-course measurements of radionuclide motion within plants.

Nondestructive real-time radioisotope imaging system for visualizing 14C-labeled chemicals supplied as CO2 in plants using Arabidopsis thaliana

We have developed a real-time radioisotope imaging system (RRIS) that can nondestructively trace 14C-labeled chemicals in plants. In an experiment, after feeding 14CO2 to a plant, the plant was fixed inside a box where lighting was regulated, and beta rays emitted from the 14C in the plant were intermittently imaged using the developed system. As a first step, using a series of standard sources of 14C, the data depth and detection limits of the 14C images captured by the RRIS were evaluated for various integral times. As a result, the linearity between the 14C activity and signal intensity was determined for the range 103. Next, the linearity was validated using plant (Arabidopsis thaliana) organs, resulting that the linearity was shown in the case of young leaf, but was not maintained in the thick organs, such as a flower, mature leaf, siliques, and stem. Considering the good correlation between the intensity by RRIS and the PSL value by an imaging plate (IP) as well as the relative low energy of beta rays emitted from 14C, the thickness of the organs would easily affect the quantitativity of the RRIS as well as an IP. Our findings prove that sequential images of 14C in a living plant sample in a regulated light and air environment can be nondestructively analyzed using the developed system, whose quantitativity is similar to that of an IP.

Development of a 14C detectable real-time radioisotope imaging system for plants under intermittent light environment

A new real-time radioisotope imaging system (RRIS) to study the kinetics of nutrient uptake and transfer of photosynthetic products in a living plant was developed and evaluated through a test run. 14C is a common radioisotope of carbon and useful to trace the photosynthetic products as well as a low energy beta emitter. The rationale of this study was to develop a RRIS that has the ability to detect low energy beta emitters, such as 14C, 35S, and 45Ca. To achieve compatibility between the detection of low energy beta emitters and irradiation of the test plant, an intermittent lighting system was added to the RRIS. Furthermore, a commercially available digital camera was added to the RRIS for acquisition of photographic images of the test plants. The capabilities of the new RRIS were evaluated through a test run by using seedlings of rice plants and 35S-labeled sulfate. It was shown that the new RRIS was able to detect 35S absorbed by rice plant seedlings, and it was able to acquire photon-counting images and photographic images of the test plants simultaneously. Despite some limitations, the new RRIS provides a means to study the kinetics of elements in plants by utilizing low energy beta emitters.

Development of real-time radioisotope imaging systems for plant nutrient uptake studies

Ionic nutrition is essential for plant development. Many techniques have been developed to image and (or) measure ionic movement in plants. Nevertheless, most of them are destructive and limit the analysis. Here, we present the development of radioisotope imaging techniques that overcome such restrictions and allow for real-time imaging of ionic movement. The first system, called macroimaging, was developed to visualize and measure ion uptake and translocation between organs at a whole-plant scale. Such a device is fully compatible with illumination of the sample. We also modified fluorescent microscopes to set up various solutions for ion uptake analysis at the microscopic level. Both systems allow numerical analysis of images and possess a wide dynamic range of detection because they are based on radioactivity.

A study of 32P-phosphate uptake in a plant by a real-time RI imaging system

Abstract It is very important to visualize the process of nutrient absorption and distribution to study the physiological activity of the plant. We developed a real-time radioisotope (RI) imaging system, where RI tracers were applied to the plant sample. This system allowed the quantitative measurement concerning the uptake of nutrients labeled with radioisotopes, such as 45Ca, 35S, 32P and 14C as long as several days. The β-rays emitted from the sample were converted to light by a CsI(Tl) scintillator and were guided to a highly sensitive CCD camera. The scintillator surface was covered with an Al plate to avoid LED light penetration but allow selected β-ray penetration. We employed Lotus japonicus for the plant sample and observed the 32P-phosphate absorption in roots and the accumulation to the aboveground part of the plant. The environment condition of daytime and night was simulated by the ON/OFF of LED timer and the accumulation manner of the 32P-phosphate in roots and leaves during daytime and night was analyzed. The accumulation of 32P-phosphate in leaves was highly dependant on light irradiation and higher when the LEDs was turned on, whereas the absorption of 32P-phosphate in root was higher when the LEDs was turned off. The transfer function concerning the transportation of phosphate within the plant during the developmental stage was obtained from the analysis of 32P uptake images. We are now trying to get specific moving images of each radioisotope when two kinds of isotopes, such as 32P and 35S, were applied at the same time to the plant, through an image analysis.