鈴井 伸郎; 丸山 哲平*; 河地 有木; 三輪 睿太郎*; 樋口 恭子*; 藤巻 秀
JAEA-Review 2015-022, JAEA Takasaki Annual Report 2014, P. 95, 2016/02
Common reed, which is classified to the same family as rice, is a salt-tolerant plant. In this study, we conducted non-invasive imaging of Na dynamics in intact common reed plants by using Na tracer and a positron-emitting tracer imaging system (PETIS) in order to understand the salt-tolerant mechanism in common reed. Common reeds and rice plants were cultivated in a nutrient solution containing 50 mM NaCl. After Na was fed into the solution, we observed the Na movement from the solution to the shoot for 24 h using PETIS. As a result, Na was strongly accumulated in the shoot base but not transported to the upper shoot in common reed, whereas Na was continuously transported to the upper shoot in rice plant. Furthermore, we replaced the original solution with a fresh nutrient solution without Na and traced the Na movement inside the plants for 18 h. Detailed quantitative analysis of the image data revealed that Na migrated downward from the shoot base to the root tip in common reed. These results indicate that common reed has constitutive ability of Na exclusion only in the direction of root tips, and consequently keeps low Na concentration in the upper shoot.
尹 永根; 河地 有木; 鈴井 伸郎; 石井 里美; 吉原 利一*; 渡部 浩司*; 山本 誠一*; 藤巻 秀
JAEA-Review 2015-022, JAEA Takasaki Annual Report 2014, P. 112, 2016/02
Large areas of agricultural fields were contaminated with radiocesium (Cs and Cs) in Japan by the accident of The Tokyo Electric Power Company Fukushima Daiichi Nuclear Power Station in March 2011. Many agricultural studies, such as fertilizer management and plant breeding, are undertaken for reducing radiocesium uptake in crops or enhancing of uptake and transportation via phytoremediation. These studies examine the control of radiocesium transport into/within plant bodies from the viewpoint of plant physiology. Radiotracer imaging is one of the few methods that enable the observation of the movement of substances in a living plant, like a video camera, without sampling of the plant tissues. In this study, we performed the imaging of Cs uptake and transport from root to aerial part by using a new gamma camera in intact soybean plants because contamination of soybean by radiocesium has currently become a major problem in agriculture in Fukushima.
河地 有木; 尹 永根; 鈴井 伸郎; 石井 里美; 吉原 利一*; 渡部 浩司*; 山本 誠一*; 藤巻 秀
JAEA-Review 2015-022, JAEA Takasaki Annual Report 2014, P. 94, 2016/02
We developed an original gamma camera system to image radiocesium in a plant. The gamma camera was designed for high-energy gamma photons from Cs radiocesium (662 keV). We performed tests to evaluate the position resolution and quantitative linearity of the gamma camera. The best spatial resolution of this gamma camera was determined to be 19.1 mm in full width at half maximum at the center of the field-of-view. And a result shows a quantitative linearity of the image data with a correlation of = 0.9985 between the source activity and the count rate. We conclude the gamma camera system has sufficiently high capability to obtain quantitative and dynamic images of Cs movement in intact plants.
石井 里美; 山崎 治明*; 鈴井 伸郎; 尹 永根; 河地 有木; 島田 浩章*; 藤巻 秀
JAEA-Review 2015-022, JAEA Takasaki Annual Report 2014, P. 93, 2016/02
Tomato is popularly grown in environmentally controlled system such as a greenhouse for improvement of bioproduction. It is important to control the condition in the greenhouse for increasing the translocation of fixed carbon from the leaves to the growing fruits. Elevation of COconcentration is widely employed for that purpose; however, it is difficult to estimate its effect quantitatively because tomato plants have too large inter-individual variations with developing fruits. In this study, we employed a PETIS which is a live-imaging system of nutrients in plant body using short-lived radioisotopes including C. We also established a closed cultivation system to feed a test plant with CO at set concentrations of 400, 1,500 and 3,000 ppm and a pulse of CO.
河地 有木; 尹 永根; 鈴井 伸郎; 石井 里美; 吉原 利一*; 渡部 浩司*; 山本 誠一*; 藤巻 秀
Journal of Environmental Radioactivity, 151(Part 2), p.461 - 467, 2016/01
We developed a new gamma camera specifically for plant nutritional research and successfully performed live imaging of the uptake and partitioning of Cs in intact plants. The gamma camera was specially designed for high-energy photons from Cs (662 keV). To obtain reliable images, a pinhole collimator made of tungsten heavy alloy was used to reduce penetration and scattering of photons. The array block of the GAGG scintillator was coupled to a high-quantum efficiency position sensitive photomultiplier tube to obtain accurate images. The completed gamma camera had a sensitivity of 0.83 count s MBq for Cs, and a spatial resolution of 23.5 mm. We used this gamma camera to study soybean plants that were hydroponically grown and fed with 2.0 MBq of Cs for 6 days to visualize and investigate the transport dynamics in aerial plant parts. Cs gradually appeared in the shoot several hours after feeding, and then accumulated preferentially and intensively in growing pods and seeds; very little accumulation was observed in mature leaves. Our results also suggested that this gamma-camera method may serve as a practical analyzing tool for breeding crops and improving cultivation techniques resulting in low accumulation of radiocesium into the consumable parts of plants.
藤巻 秀; 丸山 哲平*; 鈴井 伸郎; 河地 有木; 三輪 睿太郎*; 樋口 恭子*
Plant & Cell Physiology, 56(5), p.943 - 950, 2015/05
山崎 治明*; 鈴井 伸郎; 尹 永根; 河地 有木; 石井 里美; 島田 浩章*; 藤巻 秀
Plant Biotechnology, 32(1), p.31 - 37, 2015/04
To maximize fruit yield of tomatoes cultivated in a controlled, closed system such as a greenhouse or a plant factory at a limited cost, it is important to raise the translocation rate of fixed carbon to fruits by tuning the cultivation conditions. Elevation of atmospheric CO concentration is a good candidate. In this study, we employed a positron-emitting tracer imaging system (PETIS), which is a live-imaging technology for plant studies, and a short-lived radioisotope C to quantitatively analyze immediate responses of carbon fixation and translocation in tomatoes in elevated CO conditions. We also developed a closed cultivation system to feed a test plant with CO at concentrations of 400, 1500 and 3000 ppm and a pulse of CO. As a result, we obtained serial images of C fixation by leaves and subsequent translocation into fruits. Carbon fixation was enhanced steadily by increasing the CO concentration, but the amount translocated into fruits saturated at 1500 ppm on average. The translocation rate had larger inter-individual variation and showed less consistent responses to external CO conditions compared with carbon fixation.
樋口 恭子*; 金井 雅武*; 土屋 将久*; 石井 春香*; 渋谷 尚史*; 藤田 直子*; 中村 保典*; 鈴井 伸郎; 藤巻 秀; 三輪 睿太郎*
Frontiers in Plant Science (Internet), 6, p.138_1 - 138_6, 2015/03
In a previous study, we reported that the common reed accumulates water-soluble Cd complexed with an -glucan-like molecule, and that the synthesis of this molecule is induced in the stem of the common reed under Cd stress. We studied the metabolic background to ensure -glucan accumulation under the Cd stress conditions that generally inhibit photosynthesis. We found that the common reed maintained an adequate CO assimilation rate, tended to allocate more assimilated C to the stem, and accumulated starch granules in its stem under Cd stress conditions. AGPase activity, which is the rate-limiting enzyme for starch synthesis, increased in the stem of common reed grown in the presence of Cd. Starch accumulation in the stem of common reed was not obvious under other excess metal conditions. Common reed may preferentially allocate assimilated carbon as the carbon source for the formation of Cd and -glucan complexes in its stem followed by prevention of Cd transfer to leaves acting as the photosynthetic organ. These responses may allow the common reed to grow even under severe Cd stress conditions.
鈴井 伸郎; 河地 有木; 石井 里美; 尹 永根; 岩崎 郁*; 小川 健一*; 藤巻 秀
JAEA-Review 2014-050, JAEA Takasaki Annual Report 2013, P. 105, 2015/03
In this study, we developed an analytical method to evaluate velocity of photoassimilate flow using C-tracer and the positron-emitting tracer imaging system (PETIS). CO gas tracer was fed to the compound leaves of the soybean plant, and serial images of C distribution were obtained by PETIS. Regions of interests (ROIs) were set in the node of the first compound leaf (ROI-1) and the stem base (ROI-2). Time course of C-radioactivity (Time-Activity Curve: TAC) in each ROI was generated from the serial images. Initial slope of the rising C-radioactivity was estimated by a linear least-square method using the TAC data. The value of the intercept of approximated line to the background line (X-intercept) was defined as "C-arrival time" to the ROIs. In order to determine X-intercept with a fair criterion, we developed an analytical program. The velocities of C-photoassimilate flows of 18 individual soybean plants were estimated by the values of C-arrival time and the distance between ROI-1 and ROI-2. As a result, the average value of the velocity was 113 cm h and the standard deviation was 20 cm h. This result indicates the newly developed method is a reliable tool for the quantitative analysis of photoassimilate flow through the phloem.
山本 誠一*; 緒方 良至*; 河地 有木; 鈴井 伸郎; 尹 永根; 藤巻 秀
Nuclear Instruments and Methods in Physics Research A, 777, p.102 - 109, 2015/03
After the nuclear disaster in Fukushima, radiocesium contamination became a serious scientific concern and research of its effects on plants increased. In such plant studies, high resolution images of radiocesium are required without contacting the subjects. Cherenkov light imaging of beta radionuclides has inherently high resolution and is promising for plant research. Since Cs and Cs emit beta particles, Cherenkov light imaging will be useful for the imaging of radiocesium distribution. Consequently, we developed and tested a Cherenkov light imaging system. We used a high sensitivity cooled charge coupled device (CCD) camera for imaging Cherenkov light from Cs and Cs. A bright lens was mounted on the camera and placed in a black box. With a 100-m Cs point source, we obtained 220-m spatial resolution in the Cherenkov light image. With a 1-mm diameter, 320-kBq Cs point source, the source was distinguished within 2-s. We successfully obtained Cherenkov light images of a plant whose root was dipped in a Cs solution, radiocesium-containing samples as well as line and character phantom images with our imaging system. Cherenkov light imaging is promising for the high resolution imaging of radiocesium distribution without contacting the subject.
中村 進一*; 近藤 ひかり*; 鈴井 伸郎; 尹 永根; 石井 里美; 河地 有木; 頼 泰樹*; 服部 浩之*; 藤巻 秀
Molecular Physiology and Ecophysiology of Sulfur, p.253 - 259, 2015/00
Glutathione is a sulfur-containing peptide involved in various aspects of plant metabolism. Glutathione is also known to have effects on heavy metal responses in plants. In our previous work, we have found glutathione, applied to roots site- specifically, inhibited cadmium (Cd) translocation from roots to shoots and Cd accumulation in shoots in oilseed rape plants. In addition, we succeeded in visualizing inhibition of root-to-shoot translocation of Cd by using a positron-emitting tracer imaging system (PETIS). In this work, the effects of glutathione concentration in the root zone (hydroponic solution) and the glutathione treatment period on Cd partitioning in oilseed rape plants were investigated. Our experimental results demonstrated that glutathione, exceeding a certain concentration in the root zone, is needed to trigger inhibition of Cd translocation, and that treatment time from the start of glutathione application had different effects on Cd partitioning in oilseed rape plants.
高橋 竜一*; 石丸 泰寛*; Shimo, H.*; Bashir, K.*; 瀬野浦 武志*; 杉本 和彦*; 小野 和子*; 鈴井 伸郎; 河地 有木; 石井 里美; et al.
PLoS ONE (Internet), 9(6), p.e98816_1 - e98816_7, 2014/06
Previously, we reported that OsNRAMP5 functions as a manganese, iron, and cadmium (Cd) transporter. The shoot Cd content in 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 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 Cd 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 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.
吉原 利一*; 鈴井 伸郎; 石井 里美; 北崎 真由*; 山崎 治明*; 北崎 一義*; 河地 有木; 尹 永根; 七夕 小百合*; 橋田 慎之介*; et al.
Plant, Cell & Environment, 37(5), p.1086 - 1096, 2014/05
Cadmium (Cd) accumulations in a Cd hyper-accumulator fern, (), and tobacco, (), were kinetically analysed using the positron-emitting tracer imaging system under two medium conditions (basal and no-nutrient). In , maximumly 50% and 15% of the total Cd accumulated in the distal roots and the shoots under the basal condition, respectively. Interestingly, a portion of the Cd in the distal roots returned to the medium. In comparison with , a little fewer Cd accumulations in the distal roots and clearly higher Cd migration to the shoots were observed in under the basal condition (maximumly 40% and 70% of the total Cd, respectively). The no-nutrient condition down-regulated the Cd migration in both species, although the regulation was highly stricter in than in (almost no migration in and around 20% migration in ). In addition, the present work enabled to estimate physical and physiological Cd accumulation capacities in the distal roots, and demonstrated condition-dependent changes especially in . These results clearly suggested occurrences of species-/condition-specific regulations in each observed parts. It is probable that integration of these properties govern the specific Cd tolerance/accumulation in and .
鈴井 伸郎; 尹 永根; 井倉 将人*; 石井 里美; 河地 有木; 石川 覚*; 藤巻 秀
JAEA-Review 2013-059, JAEA Takasaki Annual Report 2012, P. 101, 2014/03
We have conducted noninvasive imaging of cadmium (Cd) in intact plants using a positron-emitting tracer imaging system (PETIS) and Cd for the purpose of understanding the mechanism of Cd translocation in plants. In our typical imaging experiment, Cd tracer has been fed to the root of large plants such as rice, and obtain the serial images of Cd distribution in the underground parts (roots and culture solutions) or the aerial parts (shoots or grains). However, whole-plant images of Cd distribution in both underground and aerial parts have not been obtained because the field of view (FOV) of PETIS is limited. In this study, we fed Cd to dwarf plants, and visualized Cd dynamics in a whole plant within the FOV of PETIS. As a result, the whole physiological process, i.e., uptake from culture solution, translocation from root to shoot and accumulation in shoot, were successfully visualized in the single imaging experiment. The time courses of Cd amounts demonstrated that the half amount of Cd fed to the solution was taken up by the root within 2 hours, and the two-thirds amount was accumulated in the shoot after 30 hours. These results imply the whole-plant imaging represents a reliable tool for the quantitative analysis of Cd dynamics.
石井 里美; 井倉 将人*; 尹 永根; Hung, N. V. P.*; 鈴井 伸郎; 河地 有木; 小柳 淳*; 大山 卓爾*; 藤巻 秀
JAEA-Review 2013-059, JAEA Takasaki Annual Report 2012, P. 98, 2014/03
Nitrogen is the most important nutrient for the plants. Soybean can utilize nitrogen from atmospheric N fixed by nodules which are symbiotic organs of leguminous plants with rhizobia. In the beginning of this research project, we set our methodological goal to visualize and analyze the nitrogen fixation in the nodules and subsequent nitrogen transport to the aerial part in a plant using N and PETIS. Previously, we have developed a method of production of highly purified N-labelled nitrogen gas tracer using gas chromatography and successfully visualized nitrogen fixation in intact nodules. However, the yields of the tracer were only a few ten megabecquerels and too low to visualize the transport of fixed nitrogen to the aerial part. Therefore, we have been trying to improve the production method to gain much higher radioactivity with consideration of the very short half-life of [N]N. In this study, we tested a new technique to turn the main by-product [N]NO into the desired product [N]N directly.
Hung, N. V. P.; 石井 里美; 鈴井 伸郎; 河地 有木; 尹 永根; 小柳 淳; 大山 卓爾*; 藤巻 秀
JAEA-Review 2013-059, JAEA Takasaki Annual Report 2012, P. 99, 2014/03
ダイズの根粒における共生的窒素固定に対する周辺空気の組成、とくに酸素分圧の及ぼす影響について解析するために、ポジトロンイメージング(PETIS)による画像化と解析を行った。窒素13ガスを製造、精製し、所定の酸素分圧となるようにトレーサガスを調製した。これをダイズ根粒に10分間投与し、PETISによりイメージングを行った。減衰を待ち、同一個体を対象に、酸素分圧を変えて(0%, 10%, 20%)実験を繰り返し、合計3回の実験を行った。各実験回において根粒が窒素を固定する動画像が得られ、これらのデータを解析したところ、酸素分圧が通常の20%から10%に低下した場合、窒素固定活性も顕著に低下するが、その程度は0%の場合とさほど変わりないことが明らかになった。
河地 有木; 尹 永根; 鈴井 伸郎; 石井 里美; 渡部 浩司*; 山本 誠一*; 藤巻 秀
JAEA-Review 2013-059, JAEA Takasaki Annual Report 2012, P. 100, 2014/03
Because of the accident at the Fukushima Daiichi Nuclear Power Plant of the Tokyo Electric Power Company, Inc., an extensive area of agricultural fields was contaminated with radioactive materials. Cs-137 is the most of soil contaminant, which was reported to have a 16 - 18 years half-life in agricultural fields affected by physical decay and soil erosion. Therefore, further research is required on the behavior of radiocesium and its transfer from contaminated soil to agricultural products. In this study, we have developed a gamma camera for ray imaging of Cs-137 emitting at 662 keV. A pinhole collimator was fabricated with heavy metal of tungsten to avoid the penetration and scattering of rays, since high-energy incident ray originates from Cs-137 tracer inside a test plant. A gadolinium oxyorthosilicate (GSO) scintillator and a flat panel position sensitive photomultiplier tube were adapted to the gamma camera to obtain adequate high sensitivity. Giant knotweed, which has potential as a cleanup plant with high uptake capacity for cesium, was grown in hydroponic solutions. After exposed to approximately 400 kBq of Cs-137, images were taken for 15 h. Sequential images reveal the changing distribution of cesium into the plant from the hydroponic solutions via the root system. We have indicated real-time visualization of uptake of radiocesium within an intact plant in the first time successfully.
Hu, P.*; 尹 永根; 石川 覚*; 鈴井 伸郎; 河地 有木; 藤巻 秀; 井倉 将人*; Yuan, C.*; Huang, J.*; Li, Z.*; et al.
Environmental Science and Pollution Research, 20(9), p.6306 - 6316, 2013/09
, a species in the family Crassulaceae, has been reported to be a Cd hyperaccumulator exhibiting fast growth, large biomass, asexual reproduction and perennial habit and showing remarkable potential in the phytoextraction of Cd from polluted soils. The aims of this study are to investigate whether and how the nitrogen form (nitrate (NO) versus ammonium (NH)) influences cadmium (Cd) uptake and translocation and subsequent Cd phytoextraction by the . This is the first report of successful visualisation and quantification of Cd uptake and root-to-shoot translocation in intact plants of the hyperaccumulator species supplied with different forms of inorganic N using positron-emitting tracer imaging system (PETIS), a real-time imaging method.
中村 進一*; 鈴井 伸郎; 長坂 俊紀*; 小松 史弥*; 石岡 典子; 伊藤 小百合*; 河地 有木; 頼 泰樹*; 服部 浩之*; 茅野 充男*; et al.
Journal of Experimental Botany, 64(4), p.1073 - 1081, 2013/02
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 () 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.
尹 永根; 鈴井 伸郎; 河地 有木; 石井 里美; 山崎 治明; 小柳 淳*; 藤巻 秀
JAEA-Review 2012-046, JAEA Takasaki Annual Report 2011, P. 92, 2013/01
The root of higher plant has important role in absorb essential nutrients critical to life. On the other hand, the root evolved special abilities to uptake of nutrients from the rhizosphere environment because that is fixed in the soil. As one example, the roots secrete organic acids to surrounding of rhizosphere for solubilization of the insoluble mineral in soil and absorb directly or indirectly of the nutrition. Previously, our group has reported that imaging of cadmium (Cd) uptake from hydroponic culture solution to root for study the mechanism of mineral metabolism by using a positron-emitting tracer imaging system (PETIS) in plant. In this study, we performed the imaging of organic matter which is exudate from root to soil cultivation by using carbon-11-labeled carbon dioxide (CO) gas tracer with PETIS.