Nitrogen-fixing microoganisms in paddy soils XVI

1. It was proved that under aerobic shaking conditions of the system of R. capsulatus mixed with Az. agilis or R. capsulatus mixed with B. megaterium, the bacterial growth of both and the nitrogen fixation were promoted markedly, comp:ired with the non-shaking culture. However, in the system of R. capsulatus mixed with B. subtilis under aerobic shaking conditions. such remarkable stimulative effects were not recognized altough some slow effects were recognized in the nonshaking culture.

2. It was also proved that under aerobic shaking conditions of the system of R. capsulatus mixed with Az. agilis or R. capsulatus mixed with B. megaterium the assimilative activity of carbon dioxide increased over 10 times more than in the separate pure culture.

From these results, it is conceivable that a very good environment was created for the internal metabolism of R. capsulatus in such mixing systems (R-Az and R-Bm).     

Nucleic acids in the culture fluids of Rhodopseudomonas capsulatus and other heterotrophic bacteria

In previous papers (1,2), it was reported that in cultures of R. capsulatus mixed with other heterotrophic bacteria nitrogen fixation is accelerated and there is some production of slime substances (3), exchange of pyruvic acid (4–6), amino acids, carbohydrates (7) and others in these mixed systems. There is also symbiotic association between R. capsulatus and other heterotrophic bacteria (Azotobacter) in nature (8).     

Prediction of grain protein content in winter wheat through leaf color measurements using a chlorophyll meter

A field trial was conducted over a 3-year period at the Hokkaido Kitami Agricultural Experiment Station to examine whether the grain protein content (GPC) of a winter wheat cultivar (Triticum aestivum L. cv. Chihokukomugi) suitable for Japanese noodle-making could be predicted before harvest. The prediction of the GPC was accurate based on the color of the second leaf (just below the flag leaf) at the end of the emergence of the inflorescence, when nitrogen application was graded. In order to evaluate the reliability of this test, a survey of 95 wheat fields in the eastern part of Hokkaido was also carried out during a 3-year period. The prediction of the GPC for this cultivar based on the color of the second leaf was less accurate across many sites. The results of this survey, however, suggested that the leaf color could be used as an index for ranking the GPC as low or high in relation to processing requirements. When the leaf color value of the second leaf measured with a chlorophyll meter at the end of the emergence of the inflorescence was less than 40, it was predicted that the GPC would be lower than the processing requirement. This index could be applied to the cultivars grown in the eastern part of Hokkaido, except for those grown on peat soils.     

Arsenic accumulation and speciation in Japanese paddy rice cultivars

We examined arsenic (As) accumulation and speciation in the major cultivars currently grown in Japan, because differences in grain As levels among cultivars may influence dietary As exposure in Japanese people. Ten major cultivars (Oryza sativa L.) were grown under flooded conditions in a paddy field with a background level of As (low-As soil) or in pots filled with soil containing a high level of As (high-As soil). In the low-As soil, the total grain As ranged from 0.11 to 0.17?mg?kg^?1, with a mean concentration of 0.14?mg?kg^?1, and inorganic As was the major species in all cultivars. There were few genotypic differences in the levels of either total As or inorganic As in the grain. In the high-As soil, total grain As increased to a mean level of 2.4?mg?kg^?1 in the 10 cultivars, with markedly increased levels of dimethylarsinic acid. The genotypic variations among cultivars in the levels of both total As and dimethylarsinic acid were statistically significant. However, the genotypic variability of inorganic As levels was quite small, and these levels remained low (at about 0.2?mg?kg^?1) even when total As levels increased markedly. These results suggest that differences in grain As levels among Japanese cultivars may not influence dietary As exposure, because there is little genotypic difference in the accumulation of inorganic As, which is considered more toxic than organic As to humans. We discuss the possible mechanism of As accumulation in Japanese paddy rice, in the context of the accumulation of As species in the developing grain and in other plant tissues.     

Evaluation of fresh pasta-making properties of extra-strong common wheat (Triticum aestivum L.)

The relationship between characterictics of flour of common wheat varieties and fresh pasta-making qualitites was examined, and the fresh pasta-making properties of extra-strong varieties that have extra-strong dough were evaluated. There was a positive correlation between mixing time (PT) and hardness of boiled pasta, indicating that the hardness of boiled pasta was affected by dough properties. Boiled pasta made from extra-strong varieties, Yumechikara, Hokkai 262 and Hokkai 259, was harder than that from other varieties and commercial flour. There was a negative correlation between flour protein content and brightness of boiled pasta. The colors of boiled pasta made from Yumechikara and Hokkai 262 grown under the condition of standard manuring culture were superior to those of boiled pasta made from other varieties. Discoloration of boiled pasta made from Yumechikara grown under the condition of heavy manuring culture was caused by increase of flour protein content. On the other hand, discoloration of boiled pasta made from Hokkai 262 grown under the condition of heavy manuring culture was less than that of boiled pasta made from Yumechikara. These results indicate that pasta made from extra-strong wheat varieties has good hardness and that Hokkai 262 has extraordinary fresh pasta-making properties.     

Isolation and structural characterization of rhamnogalacturonan II-borate complex fromPinus densiflora

A boron-containing pectic polysaccharide was isolated from Driselase digests of akamatsu (Pinus densiflora) cell wall. The boron-containing polysaccharide was purified by successive column chromatography of Cosmosil C₁₈ OPN, DEAE Sepharose FF, and Superdex 75. The complex had 0.12% (w/w) boron (B) and¹¹B-NMR spectroscopy showed the complex to be the tetrahedral diester form of boron. The sugar composition and glycosyl linkage analyses showed that the sugar portion of the complex was rhamnogalacturonan-II (RG-11). The boron-attached glycosyl residue was identified to be 2-O-Me-xylose-containing apiosyl side chain in RG-II. These results established that the location of borate in the cell wall is conserved among dicots, gramineous monocots, and coniferous gymnosperm.Part of this study was presented at the 8th Cell Wall Symposium, Norwich, September 3, 1998     

Effect of presowing seed treatments on germination and early seedling growth of wheat varieties under saline conditions

The effect of presowing wheat seed treatments was investigated for the salt-sensitive variety Blue Silver and relatively salt-tolerant variety PARI-73 at the germination and early seedling stages. Seeds were treated with distilled water (DW) or 10/50 mm KCl, KNO₃ , CaCl₂, Ca(NO₃)₂ and then germinated in DW or 200 mm NaCl. Treatment with calcium (Ca) or potassium (K) did not lead to a significantly higher rate for final germination than the DW treatment. Ca salt treatments significantly improved shoot growth during the early seedling establishment stage in both varieties, especially in the salt-sensitive variety Blue Silver. There were significant differences in the Ca content of seeds after various presowing treatments. There were also significant differences between both varieties in the ion contents after seed treatment. However, these differences appeared to be related to the improvement of shoot growth during the early seedling establishment stage and not to the effectiveness of pres owing seed treatments in increasing germination.     

Precipitation chemistry in Japan 1989–1993

Precipitation chemistry in Japan was discussed on a wet-only sample database obtained in a nationwide survey from April 1989 to March 1993. Wet-only samples were collected at 29 stations over Japan on a biweekly basis. Commonly determined chemical parameters were measured in laboratories. The volume-weighted annual mean pH at each site ranged from 4.50 to 5.83 with a mean of 4.76. Concentration ranges and means (parenthesized) on an equivalent basis for major ions were as follows: nss-SO₄ ^2–; 5.2–58.9 (38.6), NO₃ ^–; 1.8–25.0 (14.1), NH₄ ⁺; 0.55–29.8 (18.3), nss-Ca²⁺; 2.0–34.5(14.2), Na⁺; 6.4–275.3 (49.1), Cl^–; 13.7–322.4 (63.5) eq L^–1. Acid-base relationships for Phase-II records were quantitatively discussed in terms of three measures: pH, fractional acidity, and our proposed pA_i.     

Antioxidant constituents in distillation residue of Awamori spirits

Constituents in a distillation residue of Awamori (millet spirits) and their antioxidant activity are investigated in this study. The supernatant of the distillation residue obtained by centrifugation was partitioned with n-hexane, chloroform, ethyl acetate, and n-butanol against water to afford the corresponding solubles. Among them, n-hexane and chloroform solubles showed higher antioxidant potency than l-ascorbic acid by the bleomycin-Fe method. In chloroform solubles, seven cyclic dipeptides were identified along with ethyl 2-pyrrolidione-5-carboxylate, tyrosol, and ethyl p-hydoroxyphenyllactate. Antioxidant activity of ethyl p-hydoroxyphenyllactate was 4.2 times that of l-ascorbic acid, whereas cyclic dipeptides showed activity 0.89-1.29 times as strong as that of l-ascorbic acid. On the other hand, scavenging effect of cyclic dipeptides against O(2)(-.) and OH(.) by using electron spin resonance was also investigated. In the results, cyclo(l-Ile-l-Pro) showed significantly strong inhibitory effect against OH(.) (95.4% at 2.5 x 10-3 M) and cyclo(l-Phe-l-Pro), cyclo(l-Pro-l-Val), and cyclo(l-Leu-l-Pro) inhibited OH(.) 64.9, 54.1, and 51.0%, respectively, whereas alpha-tocopherol showed 37.7% inhibition, though only a few cyclic dipeptides weakly inhibited O(2)(-.).     

Isolation and characterization of a novel immunomodulatory alpha-glucan-protein complex from the mycelium of Tricholoma matsutake in basidiomycetes

Tricholoma matsutake, a high-class edible mushroom in Japan, has been reported to have excellent biological activities, but difficulty in cultivating the fruit bodies and limited bulk availability have restricted detailed studies. We have developed a method of culturing in tanks, enabling the bulk supply of the mycelia. The preparation (CM6271) exerts modulative effects on the immune competence of mice and rats. In this study, a sodium hydroxide extract of CM6271 was defatted followed by fractionation with a combination of ion exchange chromatography and gel filtration in order to identify the components involved in the expression of the activity, and a single peak fraction (MPG-1) was obtained with reversed phase chromatography. MPG-1 was a glycoprotein (sugar:protein ratio, 94.3:5.7) with a relative molecular mass of 360 kDa, and the sugar moiety contained about 90% glucose. NMR spectra and methylation analysis revealed that the alpha-1,4-linkage was the predominant glucan linkage with alpha-1,6- and alpha-1,2-linkages in the minority. The amino acid composition in the protein moiety was rich in glutamine, alanine, asparagine, leucine, glycine, valine, serine, threonine, isoleucine, and proline. MPG-1 was resistant to degradation with amylase or protease. The oral administration of MPG-1 promoted, in a dose-dependent manner, the recovery of the mouse natural killer cell activity and serum IL-12 level that had been reduced by the loading of restraint stress. The dose of MPG-1 (25 mg/kg) required for the expression of the effect decreases to 1/12 of that of CM6271 (300 mg/kg). Furthermore, MPG-1 formed a complex with TGF-beta1 in vitro, modulating the biological activity of TGF-beta1 by binding to its active form. These results indicate that the mycelium of T. matsutake contains a novel alpha-glucan-protein complex with immunomodulatory activities.