Element interconnections in Lotus japonicus: A systematic study of the effects of element additions on different natural variants

Lotus japonicus was used to study the distribution and interconnections of 15 elements in plant tissues, including essential and non-essential elements: boron (B), sodium (Na), magnesium (Mg), potassium (K), calcium (Ca), manganese (Mn), iron (Fe), nickel (Ni), copper (Cu), zinc (Zn), arsenic (As), strontium (Sr), molybdenum (Mo), cadmium (Cd) and cesium (Cs). Large amounts of B and Ca accumulated in plant leaves, while Fe, Na, Ni, As and Cd tended to mainly occur in the roots, and Mo was the only element to accumulate in the stems. The elemental compositions within plants were severely disturbed by treatment with toxic elements. Competition between element pairs in the same group (e.g. K and Cs; Ca and Sr) was not found. Iron, Cu and Zn accumulation were induced by Cd and Ni addition. When natural variants grew in a nutrition solution with subtoxic levels of As, Cd, Cs, Ni, Mo and Sr, intriguing relationships between the elements (such as Fe, As and K; Mg and Ni; Mn and Ca) were revealed using principal-component analysis. This study on the plant ionome offers detailed information of element interactions and indicates that chemically different elements might be closely linked in uptake or translocation systems.     

Phenolic constituents in the fruits of Cinnamomum zeylanicum and their antioxidant activity

Defatted cinnamon fruit powder was successively extracted with benzene ethyl acetate, acetone, MeOH, and water. The concentrated water extract contained the maximum amount of phenolics and showed the highest antioxidant activities. Hence, it was fractionated by Diaion HP-20SS, Diaion HP-20, and Sephadex LH-20 column chromatographies. It gave five purified compounds, the purities of which were analyzed by HPLC. Compounds 1-5 were identified as 3,4-dihydroxybenzoic acid (protocatechuic acid), epicatechin-(2beta-->O-7,4beta-->8)-epicatechin-(4beta-->8)-epicatechin (cinnamtannin B-1), 4-[2,3-dihydro-3-(hydroxymethyl)-5-(3-hydroxypropyl)-7-(methoxy)benzofuranyl]-2-methoxyphenyl beta-d-glucopyranoside (urolignoside), quercetin-3-O-(6-O-alpha-l-rhamnopyranosyl)-beta-d-glucopyranoside (rutin), and quercetin-3-O-alpha-l-rhamnopyranoside by using extensive spectral studies. The antioxidant activities of purified compounds were screened for their antioxidative potential using beta-carotene-linoleate and 1,1-diphenyl-2-picrylhydrazyl model systems. All of the compounds showed antioxidant and radical scavenging activities. This is the first report of the isolation and identification of nonvolatile constituents and as well as antioxidant activities from cinnamon fruits.     

The relationship between migration speed and release date for chum salmon Oncorhynchus keta fry exiting a 110-km northern Japanese river

The relationship between release date and migration speed was examined for hatchery chum salmon Oncorhynchus keta fry exiting the Nishibetsu River in eastern Hokkaido, northern Japan so that future releases might be scheduled so that fry arrive at the ocean during periods favoring high survival. Separate marked groups of chum salmon released in early April, mid-April, and early May in 2008, late March and mid-April in 2009, and mid-April in 2010 were recaptured with a rotary screw trap 12 km above the river mouth. Chum salmon in later release groups tended to migrate downstream faster than fish in earlier release groups. Those released after mid-April arrived in the lower river on average 9 days after release, while those released before mid-April arrived on average 26–28 days after release. Most marked fish arrived in the lower river during late April to mid-May. These results suggest that chum salmon are adapted to adjust their migratory speed so as to arrive at the ocean during a relatively discrete period, presumably during a time of high productivity favoring good survival.     

Benefits of live phytoplankton, <Emphasis Type="Italic">Chlorella vulgaris</Emphasis>, as a biocontrol agent against fish pathogen <Emphasis Type="Italic">Vibrio anguillarum</Emphasis>

The ability of the probiotic bacterium Sulfitobacter to inhibit the growth of two virulent strains (psh-9019 and ATCC 43306) of the fish pathogenic bacterium Vibrio anguillarum was tested. Probiotic and pathogenic bacteria were inoculated individually or together in three different types of media. Two of the phytoplankton media tested were filtrates of phytoplankton Chlorella vulgaris cultures, either the live phytoplankton (live-CV) or a condensed phytoplankton (condensed-CV). Phytoplankton culture medium, ESM, was used as a control medium without phytoplankton. In ESM, Sulfitobacter decreased the viable cell counts of both V. anguillarum strains by tenfold. In the live-CV filtrate, V. anguillarum was eradicated by Sulfitobacter within one week. Although colony counts of strain ATCC 43306 declined during the two-week co-incubation with Sulfitobacter, its growth was not fully inhibited; however, the counts were tenfold lower than that in control ESM medium. Neither of the pathogenic V. anguillarum strains were inhibited nor eradicated by Sulfitobacter in the condensed-CV filtrate medium. Our study indicates that commercially available condensed phytoplankton can enhance the growth of V. anguillarum. Thus, the addition of live phytoplankton, including the introduction of Roseobacter clade bacteria to fish larvae tanks, leads to better biocontrol of the fish pathogen V. anguillarum.     

Attenuated mutants of Potato virus Y necrotic strain produced by nitrous acid treatment and mutagenesis-in-tissue culture methods

We produced attenuated mutants of Potato virus Y necrotic strain not only by nitrous acid treatment but also by a novel method, probably unique to plant viruses, which we call the “mutagenesis-in-tissue culture method”. This relies on the natural or experimental generation of virus sequence variants within an infected plant, and then isolating the mutants by serially cloning them in plants. A total of fifteen attenuated mutants were obtained and studied. Nucleotide and amino acid sequences of the genomes of the attenuated mutant populations were compared with those parental severe isolates, and the amino acid changes in relevant genomic regions for viral attenuation were inferred. Many of the mutations were located in the 5’ half of the genome; 65 % were located in the protein 1 (P1) and helper component proteinase protein (HC-Pro) encoding regions. Amino acid changes mostly involved simultaneous changes in two or more protein encoding regions, one of which was often in the HC-Pro encoding region. The attenuated mutants M-MY10 and N-NA10 were effective in cross-protection against the original severe isolate NTND6.     

Clinical and histopathological evaluation of Dermatophagoides farinae-induced dermatitis in NC/Nga mice orally administered Bacillus subtilis

Probiotic strains have been reported to have the ability to control allergic and inflammatory diseases. In this study, we studied the inhibitory effect of Bacillus subtilis (natto) (BS) on atopic dermatitis. The effects of continuous oral administration of BS for 4 weeks on the development of atopic dermatitis induced by Dermatophagoides farinae body antigen (DF) in NC/Nga (NC) mice were evaluated using 4 groups of mice: group (Gp) DF, DF(+) with no administration of bacteria (n=3); Gp DF/BS, DF(+) and BS(+) (n=5); and Gp PBS, DF(-) with no administration of bacteria (n=3). The mice were gavaged with 1.2 × 10(17) CFU/head of BS 6 times a week for 4 weeks, and DF was applied twice a week for 4 weeks. Histopathological examination revealed significant differences in auricular thickness between Gp DF (664.4 μm, SD=78.0) and Gp DF/BS (278.7 μm, SD = 88.8; p<0.01). The dorsal skin of Gp DF/BS (316.7 μm, SD=187.4) was significantly thinner than that of Gp DF (503 μm, SD=116.3). These results suggest that continuous oral administration of fermented food-derived bacteria (BS) can be effective in alleviating the development of skin lesions induced by DF in NC mice.     

Isolation and characterization of macrophages from a mixed primary culture of bovine liver cells

Previously, we developed a simple and efficient method to isolate liver macrophages from a mixed primary culture of adult rat liver cells. To extend the applicability of this method, we isolated macrophages from mixed primary cultures of bovine liver cells. Macrophage cells proliferated on the cell sheet of mixed bovine liver cells after 8-16d of culture. These cells were detached by shaking of the culture flasks. Subsequent transfer and brief incubation in plastic dishes resulted in selective adhesion of macrophages. After rinses with PBS, attached macrophages were harvested. More than 10(6) cells could be harvested from the culture flask at intervals of 2-3d for more than three weeks. The isolated cells were strongly positive for bovine macrophage markers, such as CD68, CD172a and Iba-1. These cells exhibited functional properties of macrophages, including active phagocytosis of polystyrene microbeads, proliferative response to recombinant bovine granulocyte-macrophage colony-stimulating factor, upregulation of specific inflammatory cytokine genes upon stimulation with lipopolysaccharide, and formation of multinucleated giant cells. The shaking and attachment method provides a simple and efficient alternative to obtain bovine liver macrophages without requiring complex equipment or specialized technical skills.     

Root-shoot interaction as a limiting factor of biomass productivity in new tropical rice lines

The objective of the present study was to identify the limiting factors in biomass productivity of new tropical rice lines (panicle weight type) by analyzing the relationship between root and shoot activity. Five field experiments using three new lines, IR65598-112-2, IR65600-42-5-2, and IR68544-29-2-1-3-1-2, and one of the highest-yielding indica rice varieties (panicle number type), IR72, were conducted at the International Rice Research Institute from 1997 to 2002. Specific absorption rate of N (SARN , mg N g^-1 root dry weight d^-1), specific stem sap exudation rate (sap exudation rate, g exudate g^-1 root dry weight 12 h^-1) and specific root respiratory rate (root respiratory rate, μmol O₂ g^-1 root dry weight h^-1) were determined as indices of root activity. Relative growth rate (RGR, mg g^-1 d^-1) was used as an index of shoot activity. Compared with IR72, the new lines showed lower RGR and SAR_N values during the early growth stages. In contrast, during the late growth stages, these activities were higher in the new lines than in IR72. The SAR_B and sap exudation rate showed a linear correlation with RGR at successive growth stages, regardless of the genotypes and growth conditions. These findings indicated that active root-shoot interaction was maintained throughout the growth period in high-yielding tropical rice, including the new lines and IR72. Therefore, it was considered that SAR_N and the sap exudation rate are useful indices of root activity regulating RGR in the new lines. However, the root respiratory rate could not be used for selection, because the relationship between RGR and the root respiratory rate changed with the growth conditions. Our findings support the hypothesis that root and shoot activities during maturation are important factors affecting the productivity of the new lines, which have not yet been able to attain the targeted yield.     

Varietal difference in radiocesium uptake and transfer from radiocesium deposited soils in the genus Amaranthus

Abstract

Within Amaranthaceae, 33 different varieties, including local varieties from Japan, were grown in 2012 in a field in the town of Iino in the Fukushima prefecture, which is located approximately 51 km north of Tokyo Electric Power Company, Fukushima Daiichi Nuclear Power Plant (FDNPP). The contamination level of the soil was 2770 ± 140 Bq kg^?1 dry weight (¹³⁴Cesium (Cs) + ¹³⁷Cs, average ± SE), and the field was also cultivated in 2011. There was a significant varietal difference in the dry weight production, radiocesium accumulation and transfer factor (TF) of radiocesium from the soil to the plant. The ratio of the lowest TF to the highest TF was approximately 3. Because the ratio of ¹³⁷Cs to ¹³³Cs was significantly positive, radiocesium seems to be absorbed in a manner similar to that of ¹³³Cs. It is suggested that the varietal difference in the behavior of radiocesium uptake mainly depends on its genetic background rather than on environmental factors.     

Changes in radiocesium concentration in a blueberry (Vaccinium virgatum Aiton) orchard resulting from radioactive fallout

We investigated changes in the radiocesium concentration in a blueberry (Vaccinium virgatum Aiton) orchard in Fukushima prefecture to clarify the radiocesium contamination for 3 years after the Tokyo Electric Power Company’s Fukushima Daiichi nuclear power plant accident occurred in mid-March 2011. In the aboveground part of blueberry bushes, the concentrations of radiocesium in branches that were directly affected by radioactive fallout were the highest among the samples investigated, and the concentrations in fruits were the lowest. The concentration of radiocesium decreased exponentially in the fruits and leaves over 3 years. The radiocesium concentrations in soils were higher in the surface layer. The amount of radiocesium in organic matter on the soil surface under the canopy greatly decreased from 2011 to 2012. The transfer factor of the radiocesium from soil to blueberry fruits decreased annually. These results suggest that radiocesium adhered directly to the aboveground organs of blueberry bushes in March 2011, and it is possible that the accumulation of radiocesium in fruits is mainly due to radiocesium transfer from the branches and trunk to fruits for several years after the nuclear power plant accident.