Abstracts of Nipon Dojo-Hiryogaku Zasshi

We investigated changes in radiocesium concentrations in a Japanese chestnut (Castanea crenata Sieold &; Zucc.) orchard in Ibaraki prefecture for 3 years after the Tokyo Electric Power Company’s Fukushima Daiichi nuclear power plant accident in March 2011. The radiocesium concentrations in the aboveground organs of Japanese chestnut trees were almost the same, while the concentration in the roots was the lowest among all the organs investigated. The concentration of radiocesium decreased exponentially for 3 years in nuts, leaves and current shoots. The radiocesium concentrations in soils were higher in the surface layer, and the trend of an annual decrease in radiocesium in the soils was similar to that of the natural decay of radiocesium. The transfer factor of radiocesium from soils to nuts of Japanese chestnut decreased annually. These results suggest that radiocesium adhered directly to the aboveground organs of Japanese chestnut trees in March 2011, and that the accumulation of radiocesium in nuts is mainly due to radiocesium transfer from the branches and trunk to nuts several years after the nuclear power plant accident.     

Protozoan predation of Escherichia coli in hydroponic media of leafy vegetables

Multiple outbreaks of food poisoning associated with fresh vegetable consumptions have occurred in many countries. Numerous reports have described human pathogenic bacteria, such as Escherichia coli O157:H7 and Salmonella spp., that can internalize into fresh vegetables via root or leaf surfaces. While attempting to obtain the threshold concentration of internalization of E. coli inoculated into hydroponic medium during vegetable cultivation, we observed a rapid decrease in E. coli numbers. In the present study, we determined that the rapid decline in E. coli was not due to a physiological change into a viable but non-culturable (VNC) state. The population crash was instead caused by true bacterial death, as the rapid descent was also confirmed by micro-colony fluorescence in situ hybridization, a culture-independent method that can detect VNC cells. We next monitored the number of E. coli inoculated into intact or filter-sterilized hydroponic medium after cultivation of various types of plants. We found that the number of E. coli in intact hydroponic medium decreased markedly, whereas the level in filter-sterilized hydroponic medium was completely unchanged. This result suggests that biotic factors were present that could be eliminated by filtering. Robust predation of E. coli by protozoa (ciliates and flagellates) was observed using fluorescently labeled bacteria incorporated into the hydroponic medium. Finally, morphological identification of flagellates by scanning electron microscopy revealed the presence of a species of Stramenopiles. These findings suggest the importance of protozoa as bacterial feeders in hydroponic systems and hence the use of these organisms as potential control agents of human pathogenic bacteria.     

Nitrogen metabolism of paddy rice at heading

The ears of paddy rice at boot stage, that is, just before heading, are being imbeded in the boot leaves' sheaths and are known to produce much guttation li:juid from their spikelets.     

Changes in fatty acid composition with age and environment in different types of peat profiles in Japan

Fatty acid composition of the crude lipid fraction of peat was investigated using several typical peat profiles in Japan. Fatty acid composition varied with the peat layers accumulated in a peat profile since 32,000 years BP. Deposition of long-range transported volcanic ash tephras also affected the composition remarkably due to the acceleration of decomposition. Fatty acid composition differed among high moor, transitional moor, and low moor peat profiles sampled in several locations in Hokkaido, Japan. The difference in the plants involved in the formation of peat was considered to be a very important factor determining the fatty acid composition. For example, arachidic acid was considered to be a good indicator for the presence of reed, while the percentage of stearic acid tended to be high in the high moor peat profiles dominated by sphagnum. As the fatty acids with a longer chain length were more stable than those with a shorter chain length, the percentage of longer fatty acids (C24 and C26) tended to increase and that of shorter fatty acids (CI4 and CI6) to decrease in the lower peat profiles. The drying of peatland also exerted a similar effect on the fatty acid composition.     

Incorporation of 14C into methionine as an intermediate for phytosiderophore biosynthesis in barley

In order to verify the precursory role of methionine (Met) in the biosynthesis of the mugineic acid family of phytosiderophores (MAs), feeding experiments of ¹⁴C-Iabeled compounds to barley (Hordeum vulgare L. cv. Minorimugi) roots grown hydroponically were conducted. When both ^l4C-Glucose (Glc) and unlabeled Met were fed to segmented roots, ¹⁴C was incorporated into Met and MAs in the roots. Molar-radioactivity of Met was higher than that of the amino-butanoic-acid unit in MAs in the roots. When ^l4C-Glc and unlabeled homoserine (Hse) were fed to decapitated roots, l4C was incorporated into Met but not into Hse. Therefore, it was considered that Hse might not be a major precursor of MAs. In addition, ¹⁴C was incorporated into Met and MAs in the roots when both ¹⁴C-glycerol (Gol) and unlabeled Met were fed to segmented roots. It is suggested that MAs may be synthesized from Glc via Met, bypassing Hse, and that the MAs biosynthesis may involve an unknown pathway associated with Gol and leading to Met.     

Increase in Iron and Zinc Concentrations in Rice Grains Via the Introduction of Barley Genes Involved in Phytosiderophore Synthesis

Increasing the iron (Fe) and zinc (Zn) concentrations of staple foods, such as rice, could solve Fe and Zn deficiencies, which are two of the most serious nutritional problems affecting humans. Mugineic acid family phytosiderophores (MAs) play a very important role in the uptake of Fe from the soil and Fe transport within the plant in graminaceous plants. To explore the possibility of MAs increasing the Fe concentration in grains, we cultivated three transgenic rice lines possessing barley genome fragments containing genes for MAs synthesis (i.e., HvNAS1, HvNAS1, and HvNAAT-A and HvNAAT-B or IDS3) in a paddy field with Andosol soils. Polished rice seeds with IDS3 inserts had up to 1.40 and 1.35 times higher Fe and Zn concentrations, respectively, compared to non-transgenic rice seeds. Enhanced MAs production due to the introduced barley genes is suggested to be effective for increasing Fe and Zn concentrations in rice grains.     

Karyotype analysis of interspecific fusants of basidiomycetes by pulsed-field gel electrophoresis

The purpose of this research was to analyze the karyotype of the interspecific fusants of twoPleurotus species. Auxotrophic mutants derived from the cultivated strain ofP. ostreatus andP. cornucopiae were used. Protoplasts were fused electrically, and the fusants were selected under auxotrophic complementation. Esterase isozyme analysis showed that several fusants had isozyme bands originating from both parental strains, and others had unilateral isozyme bands. The fusant that had expressed isozyme bands of both parental strains showed chromosomal DNA bands of both of the parental strains in pulsedfield gel electrophoresis analysis. Despite the above results, the chromosomal composition of the fusants obtained by the pulsed-field gel electrophoresis did not exhibit all of the bands of both fusion parents.     

Analysis of the electrophoretic karyotype ofFlammulina velutipes

The karyotype ofFlammulina velutipes (Curt.: Fr.) Sing. was analyzed electrophoretically using contourclamped homogeneous electric field gel electrophoresis and hybridization with DNA probes. The chromosomal DNA from the monokaryon (Fv-4K) and the dikaryon (Fv-4) were resolved into six and eight bands, respectively. The sizes of the chromosomes ranged from 1.9 to 6.0 megabase (Mb) pairs. Each of the separated bands of chromosomal DNA was identified by use of five cloned probes. The number of these chromosomes was estimated to be 6 and 12, respectively; and the size of the entire genome was estimated to be about 20.1 and 38.6Mb, respectively. From a comparison of the hybridization patterns, the existence of allelic chromosomes of different sizes was deduced in the Fv-4 strain.     

Effects of Aeration, Alum Treatment, Liming, and Organic Matter Application on Phosphorus Exchange between Soil and Water in Aquaculture Ponds at Auburn, Alabama

Application of readily-oxidizable organic substrate to laboratory soil-water systems and fish ponds caused anaerobic conditions in bottom soil and water, and concentrations of soluble reactive phosphorus (SRP) increased. Aeration of ponds increased total phosphorus (TP) concentrations by suspending soil particles in the water, but SRP concentrations declined because of increased oxy- genation of bottom water and soil, Alum [Al₂(SO₄)₃·14H₂O] treatment of ponds reduced SRP and TP concentrations in ponds, but the low concentration of alum used, 20 mg/L, had little residual effect on phosphorus concentration. Application of agricultural limestone at 0.2 kg/m² to ponds with soil pH of 5.5 and Ca²⁺ concentration of 5 mg/L did not affect SRP and TP concentration. Unless pond soils were anaerobic at their surfaces, a condition not acceptable in thermally-unstratified fish ponds, soils released little phosphorus to the water. Strong adsorption of phosphorus by soils in intensive ponds with feeding is beneficial, because removal of phosphorus by aerobic soils is a control on excessive phytoplankton growth. In fertilized ponds, phosphorus must be applied at frequent intervals to replace phosphorus removed from the water by soils.     

Purification and characterization of two anionic trypsins from the hepatopancreas of carp

SUMMARY: Two trypsins, designated as trypsin A and trypsin B, have been purified from the hepatopancreas of carp. The purification procedures consisted of ammonium sulfate fractionation, and chromatographies on DEAE-Sephacel, Ultrogel AcA54 and Q-Sepharose. Trypsin A was purified to homogeneity with the molecular mass of approximately 28 kDa, while trypsin B gave two close bands of 28.5 kDa and 28 kDa on sodium dodecylsulfate polyacrylamide gel electrophoresis both under reducing and non-reducing conditions. On native-PAGE, both trypsin A and trypsin B showed a single band. Trypsin A and trypsin B revealed optimum temperature of 40°C and 45°C, respectively, and shared the same optimum pH 9.0 using Boc-Phe-Ser-Arg-MCA as substrate. Both enzymes were effectively inhibited by trypsin inhibitors and their susceptibilities were similar. The NH₂-terminal amino acid sequences of trypsin A and trypsin B were determined to 37th and 40th amino acid residue, respectively. Their sequences were very homologous, but not identical to that of a trypsin-type serine proteinase from carp muscle and these of other trypsins. Immunoblotting test using the antibody raised against trypsin A cross-reacted with trypsin B positively.