Yong-Gen Yin

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.

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.

A kinetic analysis of cadmium accumulation in a Cd hyper-accumulator fern, Athyrium yokoscense and tobacco plants

Cadmium (Cd) accumulations in a Cd hyper-accumulator fern, Athyrium yokoscense (Ay), and tobacco, Nicotiana tabacum (Nt), were kinetically analysed using the positron-emitting tracer imaging system under two medium conditions (basal and no-nutrient). In Ay, 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 Ay, a little fewer Cd accumulations in the distal roots and clearly higher Cd migration to the shoots were observed in Nt 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 Ay than in Nt (almost no migration in Ay and around 20% migration in Nt). 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 Ay. 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 Ay and Nt.

Effects of Glutathione Concentration in the Root Zone and Glutathione Treatment Period on Cadmium Partitioning in Oilseed Rape Plants

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.

Arabidopsis thaliana FLO2 is Involved in Efficiency of Photoassimilate Translocation, Which is Associated with Leaf Growth and Aging, Yield of Seeds and Seed Quality

FLO2, FLOURY ENDOSPERM 2, is highly conserved in higher plants, and rice FLO2 has been predicted to be involved in regulation of accumulation of storage compounds. We analyzed the function of Arabidopsis thaliana FLO2 (AtFLO2) because A. thaliana set structurally different seeds from those of rice. Although the flo2 mutant of A. thaliana showed normal germination, inflorescence and morphogenesis of flowers, peculiar phenotypes on leaves and siliques were observed, suggesting that this gene played important roles during both the vegetative and reproductive stages. The mutant leaves showed a decrease in chloroplast numbers, and increased total biomass with faster growth. When grown in high light intensity conditions, it was observed that aging events were induced. The flo2 mutant showed depressed transportation of photoassimilates into the sink organs. In the reproductive stage, the flo2 mutant had significantly smaller size siliques, causing a reduced yield of seeds. These seeds were structurally weak, and the quality of seeds was significantly lowered, with reduction of accumulation of storage compounds by seeds. A positron-emitting tracer imaging system (PETIS) analysis detected a decreased amount of photoassimilate transport in the flo2 mutant. Therefore, it was presumed that the phenotypes of the flo2 mutant were caused by reduced performance of translocation or transportation of the photoassimilates. Our observation suggests that AtFLO2 is strongly involved in regulation of translocation and transport of assimilates, and contributes greatly to quality control of the various processes involving substance supply or transfer, such as photoassimilation, leaf enlargement, yield of seeds in a silique and accumulation of seed storage compounds.

Development of a Cherenkov light imaging system for studying the dynamics of radiocesium in plants

ABSTRACT High-resolution images of radiocesium (137Cs) distribution are required to study cesium kinetics in plants. A Cherenkov light imaging system can visualize fine distributions of radionuclides emitting beta particles using an optical camera. To evaluate the linearity of the system, an imaging test was performed with point sources of 137Cs, with a radioactivity of 10–2000 kBq. The results indicated that the system has a good linearity between the image intensity and the radioactivity of 137Cs. We developed an imaging system for plants using this system to study radiocesium movement in intact plants. To demonstrate the ability to image radiocesium in a plant, an experiment was performed with an intact soybean plant for four days. The root of an 11-day-old soybean plant was dipped in 20 mL of a culture solution containing 137Cs with a radioactivity of 10 MBq without potassium. After one day, the solution was replaced with one with potassium but no 137Cs. The soybean plant was in healthy condition in the system, and the high-resolution serial images indicated that 137Cs was transported to the shoot and accumulated in the node. Therefore, Cherenkov light imaging is promising for imaging radiocesium in intact plants.

Development of a low-energy high resolution X-ray camera for high-energy gamma photon background environments

ABSTRACT Although a high-energy gamma camera can obtain images of 137Cs distribution by detecting the 662-keV gamma photons, its spatial resolution is reduced because high-energy gamma photons penetrate the edge of the pinhole collimator. To solve this problem, we developed a low-energy X-ray camera that detects the characteristic X-ray photons (32–37 keV) that are emitted from 137Cs to obtain high resolution images. We used a 45 × 45 × 1-mm-thick NaI(Tl) scintillator that was encapsulated in 0.1-mm-thick aluminum and optically coupled to a 2-inch square, position sensitive photomultiplier tube (Hamamatsu Photonics, PSPMT:H12700 MOD) as an imaging detector. The imaging detector was encased in a 2-cm-thick tungsten alloy container and a pinhole collimator was attached to its camera head. The spatial resolution and sensitivity were ∼5 mm full-width at half-maximum and ∼0.6 cps/MBq for the 1.5-mm pinhole collimator 10 cm from the collimator surface, respectively. We administered 5 MBq of 137Cs to a soybean seedling, imaged the distribution of radionuclides for six hours, and successfully obtained a high resolution image of it with our developed X-ray camera. We believe our camera will be a powerful tool for such 137Cs imaging in plants.