Tomoko M. Nakanishi

Phosphate Import in Plants: Focus on the PHT1 Transporters.

The main source of phosphorus for plants is inorganic phosphate (Pi), which is characterized by its poor availability and low mobility. Uptake of this element from the soil relies heavily upon the PHT1 transporters, a specific family of plant plasma membrane proteins that were identified by homology with the yeast PHO84 Pi transporter. Since the discovery of PHT1 transporters in 1996, various studies have revealed that their function is controlled by a highly complex network of regulation. This review will summarize the current state of research on plant PHT1 multigenic families, including physiological, biochemical, molecular, cellular, and genetics studies.

Cesium adsorption/desorption behavior of clay minerals considering actual contamination conditions in Fukushima.

Cesium adsorption/desorption experiments for various clay minerals, considering actual contamination conditions in Fukushima, were conducted using the 137Cs radioisotope and an autoradiography using imaging plates (IPs). A 50 μl solution containing 0.185 ~ 1.85 Bq of 137Cs (10−11 ~ 10−9 molL−1 of 137Cs) was dropped onto a substrate where various mineral particles were arranged. It was found that partially-vermiculitized biotite, which is termed “weathered biotite” (WB) in this study, from Fukushima sorbed 137Cs far more than the other clay minerals (fresh biotite, illite, smectite, kaolinite, halloysite, allophane, imogolite) on the same substrate. When WB was absent on the substrate, the amount of 137Cs sorbed to the other clay minerals was considerably increased, implying that selective sorption to WB caused depletion of radiocesium in the solution and less sorption to the coexisting minerals. Cs-sorption to WB continued for about one day, whereas that to ferruginous smectite was completed within one hour. The sorbed 137Cs in WB was hardly leached with hydrochloric acid at pH 1, particularly in samples with a longer sorption time. The presence/absence of WB sorbing radiocesium is a key factor affecting the dynamics and fate of radiocesium in Fukushima.

Agricultural implications of the Fukushima nuclear accident.

More than 4 years has passed since the accident at the Fukushima Nuclear Power Plant. Immediately after the accident, 40 to 50 academic staff of the Graduate School of Agricultural and Life Sciences at the University of Tokyo created an independent team to monitor the behavior of the radioactive materials in the field and their effects on agricultural farm lands, forests, rivers, animals, etc. When the radioactive nuclides from the nuclear power plant fell, they were instantly adsorbed at the site where they first touched; consequently, the fallout was found as scattered spots on the surface of anything that was exposed to the air at the time of the accident. The adsorption has become stronger over time, so the radioactive nuclides are now difficult to remove. The findings of our study regarding the wide range of effects on agricultural fields are summarized in this report.

Biological effect of radiation‐degraded alginate on flower plants in tissue culture

Alginate with a weight‐average molecular mass (Mw) of approx. 9.04×105 Da was irradiated at 10–200 kGy in 4% (w/v) aqueous solution. The degraded alginate product was used to study its effectiveness as a growth promoter for plants in tissue culture. Alginate irradiated at 75 kGy with an Mw of approx. 1.43×104 Da had the highest positive effect in the growth of flower plants, namely limonium, lisianthus and chrysanthemum. Treatment of plants with irradiated alginate at concentrations of 30–200 mg/l increased the shoot multiplication rate from 17.5 to 40.5% compared with control. In plantlet culture, 100 mg/l irradiated alginate supplementation enhanced shoot height (9.7–23.2%), root length (9.7–39.4%) and fresh biomass (8.1–19.4%) of chrysanthemum, lisianthus and limonium compared with that of the untreated control. The survival ratios of the transferred flower plantlets treated with irradiated alginate were almost the same as the control value under greenhouse conditions. However, better growth was attained for the treated plantlets.

Leaf senescence in rice due to magnesium deficiency mediated defect in transpiration rate before sugar accumulation and chlorosis.

Magnesium (Mg) is an essential macronutrient supporting various functions, including photosynthesis. However, the specific physiological responses to Mg deficiency remain elusive. In this study, 2-week-old rice seedlings (Oryza sativa. cv. Nipponbare) with three expanded leaves (L2-L4) were transferred to Mg-free nutrient solution for 8 days. In the absence of Mg, on day 8, L5 and L6 were completely developed, while L7 just emerged. We also studied several mineral deficiencies to identify specific responses to Mg deficiency. Each leaf was analyzed in terms of chlorophyll, starch, anthocyanin and carbohydrate metabolites, and only absence of Mg was found to cause irreversible senescence of L5. Resupply of Mg at various time points confirmed that the borderline of L5 death was between days 6 and 7 of Mg deficiency treatment. Decrease in chlorophyll concentration and starch accumulation occurred simultaneously in L5 and L6 blades on day 8. However, nutrient transport drastically decreased in L5 as early as day 6. These data suggest that the predominant response to Mg deficiency is a defect in transpiration flow. Furthermore, changes in myo-inositol and citrate concentrations were detected only in L5 when transpiration decreased, suggesting that they may constitute new biological markers of Mg deficiency.

Inspections of radiocesium concentration levels in rice from Fukushima Prefecture after the Fukushima Dai-ichi Nuclear Power Plant accident

We summarize the inspections of radiocesium concentration levels in rice produced in Fukushima Prefecture, Japan, for 3 years from the nuclear accident in 2011. In 2011, three types of verifications, preliminary survey, main inspection, and emergency survey, revealed that rice with radiocesium concentration levels over 500 Bq/kg (the provisional regulation level until March 2012 in Japan) was identified in the areas north and west of the Fukushima nuclear power plant. The internal exposure of an average adult eating rice grown in the area north of the nuclear plant was estimated as 0.05 mSv/year. In 2012, Fukushima Prefecture authorities decided to investigate the radiocesium concentration levels in all rice using custom-made belt conveyor testers. Notably, rice with radiocesium concentration levels over 100 Bq/kg (the new standard since April 2012 in Japan) were detected in only 71 and 28 bags out of the total 10,338,000 in 2012 and 11,001,000 in 2013, respectively. We considered that there were almost no rice exceeding 100 Bq/kg produced in Fukushima Prefecture after 3 years from the nuclear accident, and the safety of Fukushimas rice were ensured because of the investigation of all rice.

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.

Highly sensitive analytical method for aluminum movement in soybean root through lumogallion staining

We present highly sensitive aluminum detection method in root using fluorescent lumogallion. Roots treated with 100 μM AlCl3 including 0.2 mM CaCl2 (pH 4.5) were stained for 60 min with 10 μM lumogallion fluorescence solution and fluorescence from aluminum complex in root was observed under confocal laser microscope. There was a good correlation between the intensity of fluorescence and aluminum content. When the amount of aluminum lost during each step in staining process was measured, it was found that about 10% of aluminum was lost only at staining stage. Through lumogallion staining method, aluminum accumulation especially at an early stage of aluminum treatment in root was shown. At the beginning (2 hr), aluminum began to be accumulated in root cap. After 4 hr treatment, the aluminum distribution was spread to about 3 mm from root apex in the root cap and outer cortex. When aluminum was found in the outer cortex in 3–5 mm from the root apex, the viability was tended to be decreased in the same area (6 hr). At the same time, aluminum amount in meristem was increased. However the comparison of lumogallion staining method with that of morin, which has been widely used to detect aluminum in root, the sensitivity of lumogallion method was found to be much higher.

Water movement in a plant sample by neutron beam analysis as well as positron emission tracer imaging system

We present water imaging of a plant sample both by neutron beam and positron emission tracer imaging system (PETIS). The former method provided static water profile in a plant sample as well as that in the vicinity of a root imbedded in soil. Not only X-ray film but also CT method using a cooled CCD camera is presented. Through non-destructive water image in an X-ray film, root development as well as 2-dimensional water movement toward the root was analyzed. Spatial water image was constructed from 180 CT projection images, taken at an interval of one degree while rotating the sample, through a CCD camera. In the case of a soybean root, there was a water gradient toward a root in soil and gave minimum value at about 1 mm far from the surface of a root. The water absorbing part in a root was gradually shifted downward with the root development. We also present real time water movement by PETIS, where water was labeled with a positron emitting nuclide, 15O. The transportation of 15O-water within a plant was relatively slow and water uptake was observed only at the lowest internode, between a root and the first leaf, during 20-minute measurement.

Biological effect of irradiated chitosan on plants in vitro.

For degradation of chitosan, chitosan with an 80% degree of deacetylation and a weight‐average molecular mass (Mw) of approx. 48 kDa was irradiated with γ‐rays at doses up to 200 kGy in a 10% (w/v) solution. The Mw of chitosan was reduced from 48 to 9.1 kDa by irradiation. The characteristics of irradiated chitosan were analysed by using Fourier‐transform IR spectroscopy and an elemental analyser. The amino group was found to be stable, whereas the C‐O‐C group decreased with increase in the dose. The product of chitosan irradiated at 100 kGy with an Mw of approx. 16 kDa showed the strongest growth promotion effect on plants in vitro. For shoot culture, supplementation with irradiated chitosan increased the fresh biomass of shoot clusters (7.2–17.0%) as well as the shoot multiplication rate (17.9–69.0%) for Chrysanthemum morifolium (florists chrysanthemum), Limonium latifolium (limonium or sea‐lavender), Eustoma grandiflorum (lisianthus, tulip gentian or Texas bluebell) and Fragaria ananassa (modern garden strawberry). The optimum concentrations of irradiated chitosan were found to be approx. 70–100 mg/l for chrysanthemum, 50–100 mg/l for lisianthus and 30–100 mg/l for limonium. For the plantlet culture, the optimum concentrations were found to be approx. 100 mg/l for chrysanthemum, 30 mg/l for lisianthus, 40 mg/l for limonium and 50 mg/l for strawberry. Supplementation with optimum concentrations of irradiated chitosan resulted in a significant increase in the fresh biomass (68.1% for chrysanthemum, 48.5% for lisianthus, 53.6% for limonium and 26.4% for strawberry), shoot height (19.4% for chrysanthemum, 16.5% for lisianthus, 33.9% for limonium and 25.9% for strawberry) and root length (40.6% for chrysanthemum, 66.9% for lisianthus, 23.4% for limonium and 22.6% for strawberry). In addition, treatment with irradiated chitosan enhanced the activity of chitosanase in treated plants and also improved the survival ratio and growth of the transferred plantlets acclimatized for 10–30 days under greenhouse conditions.