ABSTRACTS of the Articles Printed in Journal of the Science of Soil and Manure,Japan

In the preceding paper (1), it was reported that even under aerobic shaking conditions, if R. capsulatus were mixed with Bacillus megaterium, the bacterial growth of both and nitrogen fixation were accelerated remarkably.     

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).     

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.     

Effect of increased feeding of dietary α‐linolenic acid by grazing on formation of the cis9,trans11–18:2 isoform of conjugated linoleic acid in bovine milk

Feeding systems such as grazing affect the fatty acid profile of bovine milk fat. In addition, milk fat is formed as the product of fatty acid metabolism in cow bodies before being secreted into milk. However, how grazing influences milk fatty acid profile through the metabolism has not been completely characterized. When fatty acid concentrations in Holstein milk were compared between grazing and non‐grazing periods, α‐linolenic acid was significantly higher in the grazing period than in the non‐grazing period. This could be explained with an increase in α‐linolenic acid feeding with grazing. α‐linolenic acid had a linear positive correlation with conjugated linoleic acid (9c,11t‐18:2) (CLA) and vaccenic acid (VA) during the grazing period, whereas CLA had higher correlation with linoleic acid rather than with α‐linolenic acid during the non‐grazing period. These data indicate that the high content of dietary α‐linolenic acid affects CLA and VA formation in milk of grazing periods via α‐linolenic acid metabolism into VA.     

Feeding Solanum glaucophyllum to preparturient multiparous cows prevents postparturient hypocalcemia

Solanum glaucophyllum (SG) contains 1,25‐dihydroxyvitamin D₃ (1,25‐(OH)₂D₃) glycosides. We investigated the effect of SG on hypocalcemia in cows. Serum levels of 1,25‐(OH)₂D₃, total calcium and phosphorus dose‐relatedly increased after feeding with SG, while serum magnesium and chloride levels fell (P < 0.05). We also performed an ethylenediaminetetraacetic acid (EDTA) infusion to induce artificial hypocalcemia. Cows that had been fed 4.0 mg/kg body weight of SG daily for 2 weeks had a higher serum concentration of total calcium at the end of EDTA infusion than those not fed SG (P < 0.05). In a field trial, multiparous cows were assigned to one of four groups: (1) no SG, (2) 1.3 g or (3) 2.6 g of SG daily from 14 days before the estimated calving day until 3 days after calving, or (4) a single feed of 35.75 g SG at 3 days before the estimated calving day. The concentrations of serum total calcium after the calving in each treatment group were (1) 7.4, (2) 7.9, (3) 8.0 and (4) 8.9 mg/dL and higher for (4) than for (1) (P < 0.05). The data suggests that feeding a high dose of SG before the calving may maintain higher concentrations of serum calcium after the calving.     

Establishment of ES cells from inbred strain mice by dual inhibition (2i)

A number of mouse ES cells from inbred strains have been established to date, but efficiency varies across the different strains. The 129 strain mouse is efficient to establish, whereas C57BL/6 and BALB/c strains are not. It is possible that their genetic backgrounds account for the difference in their ability to establish ES cell lines. In this study, we attempted to establish C57BL/6J and BALB/c Cr ES cells by dual inhibition (2i) using two inhibitors (PD0325901 and CHIR99021) of extracellular signal regulated-kinase (ERK) and glycogen synthase kinase-3 (GSK-3), which promote ES cell differentiation. The results revealed that the establishment efficiencies of C57BL/6J and BALB/c Cr ES cells were remarkably increased by 2i. These ES cells stably expressed pluripotent markers and generated high-contribution chimeras with germline transmission. Furthermore, we generated germline chimeras from C57BL/6J ES cells through the method of gene modification. These findings indicate that 2i is a powerful tool for establishing C57BL/6J and BALB/c Cr ES cells with the ability to generate germline chimeras.     

Changes in collagen fiber content and hepatic stellate cell distribution in the liver of chick embryos and growing chickens

The content of collagen and the distribution of hepatic stellate cells (HSCs) were studied to elucidate the occurrence of sex‐dependent variations in the liver of developing embryos and growing chickens. Chick embryos from embryonic days (e) 12 to e20 and chicks at 1, 4 and 8 weeks were analyzed. Liver tissue was processed using NaOH maceration and freeze‐dried to obtain the collagen fiber specimens. HSCs were identified by double fluorescent immunohistochemistry for desmin and vimentin. There were no sex‐dependent variations in the percentage of collagen fiber per liver weight and HSC area during embryonic stages. However, the content of collagen fiber increased during embryonic development in both sexes. On the other hand, the area of HSCs significantly increased in growing males but did not show any change in females. Importantly, sex differences were observed in both collagen fiber content and HSC area in the liver at 8 weeks. These results indicate that the occurrence of collagen content variations takes place at 8 weeks in chicken liver, suggesting that a sex‐dependent hormone may play an important role on the collagen production of HSCs in the growing chicken liver.     

Invasion and colonization of mature apple fruit by Erwinia amylovora tagged with bioluminescence genes

Invasion and colonization of mature apple fruit by a transformant of Erwinia amylovora tagged with bioluminescence genes from Vibrio fischeri was examined. The transformant was deposited on cut surfaces of fruit stems, wounds on the shoulders and calyces, injured fruit-bearing twigs of harvested apple fruit, and cut fruit flesh. After incubation in closed stainless steel or plastic boxes at 25°C, fruit were periodically observed with a two-dimensional luminometer. The presence of the transformant in luminous areas was confirmed by isolating it on selective media. E. amylovora, when deposited in fruit stems: (1) can invade mature as well as immature apple fruit; (2) vertically and horizontally spreads and colonizes along vascular bundles, increasing its population; (3) reaches the calyx end and the flesh just under the exocarp within 3–4 days after inoculation; (4) when deposited on cut fruit flesh, irrespective of its maturity, can easily increase its population and survive 2–4 weeks or more at 25°C; and (5) even at the time of fruit maturation, can migrate within twigs rapidly and reaches the abscission layers between fruit-bearing twigs and fruit stems.