Growth, nitrogen fixation and mineral acquisition of Alnus sieboldiana after inoculation of Frankia together with Gigaspora margarita and Pseudomonas putida

The role of tetrapartite associations among Frankia, Gigaspora margarita (an arbuscular mycorrhizal fungus), Pseudomonas putida (rhizobacterium), and Alnus sieboldiana in growth, nitrogen fixation, and mineral acquisition of A. sieboldiana was investigated. Seedlings of A. sieboldiana were inoculated with Frankia isolated from root nodules of alder, followed by inoculation of G. margarita and P. putida, and were grown for 5 months in a greenhouse. The seedlings inoculated with Frankia and G. margarita together produced the highest biomass of shoots and root nodules. Nitrogen-fixation activity, measured by acetylene reduction assay, was observed when Frankia was inoculated. The activity, on a per-nodule gram basis, decreased after G. margarita inoculation, but on a per-plant basis there was no significant difference in the activity among inoculation treatments. The mineral content in the seedlings changed after inoculation with Frankia, but not after inoculation with P. putida and/or G. margarita. The results showed a synergistic interaction among Frankia, the mycorrhizal fungus, and the rhizobacterium on the growth of A. sieboldiana.     

Relationships of stem surface area to other stem dimensions for Japanese cedar (<Emphasis Type="Italic">Cryptomeria japonica</Emphasis> D. Don) and Japanese cypress (<Emphasis Type="Italic">Chamaecyparis obtusa</Emphasis> Endl.) trees

I investigated the relationships between stem surface area outside bark and other stem dimensions for Japanese cedar (Cryptomeria japonica D. Don) and Japanese cypress (Chamaecyparis obtusa Endl.) trees. The stem dimensions used here were the basal area and the product of diameter at breast height (dbh) and total tree height. The regression equation of the stem surface area s against basal area g was s = 184.216g for the cedar trees and s = 156.878g for the cypress trees. The slope of the equation was significantly different between the two species. For the same dbh, the cedar trees tended to have a higher total tree height than the cypress trees. The cedar trees also had a larger surface area of relative stem form than the cypress trees. These results indicated that the difference in the slope was produced by the differences in both the stem slenderness and tapering between the two species. On the other hand, the regression equation between the stem surface area and the product of dbh and total tree height dh was s = 1.937dh for the cedar trees and s = 1.921dh for the cypress trees, whereas no significant difference in the slope was found. The obtained slopes for the cedar and cypress trees seemed to be in accord with that for other coniferous species reported in earlier studies, suggesting that the variation in the slope among coniferous species would be small. The estimation from the basal area would provide a simpler means for estimating the stem surface area and would be useful in obtaining an approximation of the surface area. By contrast, the estimation from the product of dbh and total tree height would provide a more accurate and precise estimate as well as a wider applicable range, i.e., a parameter for physiological growth models. In conclusion, it could not be judged which regression equation examined in the present study was superior to the other, and thus it was important to select an appropriate equation depending both on the purpose and on the time and labor available.     

The nutrient translocation of seedlings with chemical analyses of the xylem saps ofCryptomeria japonica D. Don andPinus densiflora Sieb. et Zucc.

The chemical compositions of xylem saps ofCryptomeria japonica D. Don andPinus densiflora Sieb. et Zucc. were studied. One-year-old seedlings of these species were transplanted into hydroponics in the middle of June. Some seedlings were retained in the nursery. Two nitrogen sources, ammonium and nitrate nitrogen, were applied separately. The samplings of xylem saps were made at the end of July. Saps were collected from both roots and tops of the seedlings using a pressure chamber. InC. japonica, the fact that the translocation form of nitrogen from the roots to shoots mainly was citrulline was confirmed. The result ofP. densiflora in this study showed that glutamine was the predominant compound of nitrogen translocation. Total nitrogen ofP. densiflora had a large value in the series of ammonium. Glutamine increased in the ammonium series of both species. Nitrate nitrogen was found in whole samples ofC. japonica, and was not found inP. densiflora. As the result of cations in the saps, monovalent cations were rich inP. densiflora, and divalent cation concentrations were greater inC. japonica. The idea that the great oxygen requirement ofP. densiflora produces the negative charge gives an explanation of the great retention of divalent cations in the roots. The idea that the mineral distribution indicates root activity related to its negative charge production also is considered a possibility.     

In vivo anti-influenza virus activity of an immunomodulatory acidic polysaccharide isolated from Cordyceps militaris grown on germinated soybeans

An acidic polysaccharide (APS) was isolated from the extract of Cordyceps militaris grown on germinated soybeans. Analyses of sugar composition indicated that APS consisted of d-galactose, L-arabinose, D-xylose, L-rhamnose, and D-galacturonic acid. On the basis of the result of methylation analysis, APS was considered to be mainly composed of Araf-(1-->, -->5)-Araf-(1-->, -->4)-Galp-(1--> and -->4)-GalAp-(1--> residues. When the polysaccharide was intranasally administered, it decreased virus titers in the bronchoalveolar lavage fluid and the lung of mice infected with influenza A virus and increased survival rate. Furthermore, APS increased TNF-alpha and IFN-gamma levels in mice when compared with those of untreated mice. APS enhanced nitric oxide (NO) production and induced iNOS mRNA and protein expressions in RAW 264.7 murine macrophage cells. The induction of mRNA expression of cytokines including IL-1beta, IL-6, IL-10, and TNF-alpha was also observed. These results demonstrated that APS might have beneficial therapeutic effects on influenza A virus infection at least in part by modulation of the immune function of macrophages.     

The histopathogenesis of paralytic rabies in six-week-old C57BL/6J mice following inoculation of the CVS-11 strain into the right triceps surae muscle

A fatal encephalomyelitis was developed after intracerebral and hind limb inoculation of in 6-week-old C57BL/6J mice by the inoculation of fixed rabies virus (CVS-11 strain), intracerebrally and into hind. After the intracerebral inoculation, virus antigens were detected in the cerebral cortex and hippocampus at 2 days postinoculation (PI), and later spread centrifugally to thalamus, brain stem, cerebellum, spinal cord and spinal ganglia. At 4 days PI, severe apoptosis and DNA fragmentation were observed in the hippocampus and cerebral cortex. All mice infected intracerebrally were dead without limb paralysis at from 10 to 11 days PI. In contrast, mice infected with virus intramuscularly were persistently observed virus antigens in the myocytes at the site of inoculation from 2 days PI. At 4 days PI, the antigens were demonstrated in the spinal dorsal root ganglia, spinal cord and muscle spindles without their detection in the cerebrum and hippocampus. There were no apoptosis in the spinal cord and dorsal root ganglia, however hind limb paralysis was found in all infected mice. Hind limb paralysis was progressed to quadriparalysis, and mice were dead from 11 to 13 days PI. From 4 days PI, necrosis of neuron was observed in the the spinal and dorsal ganglia with infiltration of lymphocyte. This study suggested that the necrosis of spinal neurons was more important to cause the paralysis of hind limb rather than the severe cerebral infection and apoptosis in C57BL/6J mice infected with CVS-11 strain. The virus primarily replicated in the muscles was ascended the spinal cord via afferent fibers and retrogradely invaded the cerebrum, and with subsequent spread to muscle spindles.     

Effect of the dominant follicle aspiration before or after luteinizing hormone surge on the corpus luteum formation in the cow

Luteinizing hormone (LH) surge and follicle rupture act as trigger to start corpus luteum (CL) formation. Thus, we aimed to investigate whether a dominant follicle that has not been exposed to an LH surge can become a functional CL. For this purpose, follicular fluid from the dominant follicles (DF) of cows was aspirated before or after a GnRH-induced LH surge, and subsequent CL formation was observed. Holstein cows were divided into four groups as follows: Luteal phase, a DF was aspirated 7 days after GnRH injection; Pre-LH surge, a DF was aspirated 42 h after PGF(2alpha) injection during the mid luteal phase; Post-LH surge, a DF was aspirated 24 h after GnRH injection following PGF(2alpha); and Intact follicle, ovulation was induced by GnRH injection after PGF(2alpha). Observation of morphological changes in the aspirated follicle using color Doppler ultrasonography and blood sampling was performed on Days 0, 3, 6, and 9 (Day 0 = follicle aspiration). CL formation following DF aspiration was observed only in the Post-LH surge group. In both the Luteal phase and Pre-LH surge groups, however, none of the cows showed local blood flow at the aspirated site or CL formation. Luteal blood flow area, CL volume, and plasma progesterone concentration in the Post-LH surge group were no different from those in the Intact follicle group. The present results clearly demonstrate that rather than follicle rupture, it is the LH surge that is essential for CL formation in cows.     

Acetaldehyde emission from wood induced by the addition of ethanol

A mechanism of acetaldehyde emission from wood induced by the addition of ethanol was proposed. It is known that acetaldehyde generation is due to the oxidation of ethanol via a metabolic process involving alcohol dehydrogenase (ADH) in living bodies. However, it remains unclear whether the enzymatic alcohol oxidation is applicable to wood. We investigated possible factors of wood parts, conditioning, storage sites, and heating and sterilization treatments on acetaldehyde emission using the syringe method and HPLC analysis. We reconfirmed that acetaldehyde emission was observed only when ethanol was added to wood. Greater acetaldehyde emissions were obtained in heartwood compared to sapwood in both Japanese cedar (Cryptomeria japonica D. Don) and Japanese cypress (Chamaecyparis obtusa Endl.) specimens. In addition, an acetaldehyde conversion rate of 1–2 mol% was determined in green cedar heartwood samples, while, conversely, air-dried cedar heartwood samples showed 4–5 mol%. Ethylene oxide gas sterilization had the effect of decreasing acetaldehyde emission on air-dried wood, but not on green wood. Autoclave sterilization could completely prevent acetaldehyde emissions from both green and air-dried wood. These results suggested that an original ADH in wood and an attached ADH from the outside via microorganisms onto wood were assumed to be the primary causes of acetaldehyde emissions from wood induced by the addition of ethanol.     

Screening and genetic analysis of resistance to peanut stunt virus in soybean: identification of the putative Rpsv1 resistance gene

The peanut stunt virus (PSV) causes yield losses in soybean and reduced seed quality due to seed mottling. The objectives of this study were to determine the phenotypic reactions of soybean germplasms to inoculation with two PSV isolates (PSV-K, PSV-T), the inheritance of PSV resistance in soybean cultivars, and the locus of the PSV resistance gene. We investigated the PSV resistance of 132 soybean cultivars to both PSV isolates; of these, 73 cultivars exhibited resistance to both PSV isolates. Three resistant cultivars (Harosoy, Tsurunotamago 1 and Hyuga) were crossed with the susceptible cultivar Enrei. The crosses were evaluated in the F₁, F₂ and F_2:3 generations for their reactions to inoculation with the two PSV isolates. In an allelism test, we crossed Harosoy and Tsurunotamago 1 with the resistant cultivar Hyuga. The results revealed that PSV resistance in these cultivars is controlled by a single dominant gene at the same locus. We have proposed Rpsv1, as the name of the resistance gene in Hyuga. We also constructed a linkage map using recombinant inbred lines between Hyuga × Enrei using 176 SSR markers. We mapped Rpsv1 near the Satt435 locus on soybean chromosome 7.     

Upregulation of interferon-stimulated genes and interleukin-10 in peripheral blood immune cells during early pregnancy in dairy cows

In cows, interferon-tau (IFNT) regulates maternal recognition around days 15-19 after artificial insemination (AI). The present study hypothesized that if key target genes of IFNT are clearly upregulated in earlier stages of pregnancy, these genes could be use as indices of future pregnancy in cows. Therefore, we determined the expression of these genes in peripheral blood mononuclear leukocytes (PBMCs) and polymorphonuclear granulocytes (PMNs) during the maternal recognition period (MRP). Twenty multiparous Holstein cows were subjected to AI on day 0 and categorized into the following groups: pregnancy (Preg, n = 9), embryonic death (ED, n = 5) and non-pregnancy (NP, n = 6). Progesterone levels in the Preg group were higher than those in the NP group on days 12-21. ISG15 and OAS-1 (IFN-stimulated genes: ISGs) mRNA in PBMCs on day 8 was higher in the Preg group than in the NP group, and these mRNAs in PMNs was higher in the Preg group on day 5 than in the NP and ED groups. Interleukin-10 (IL-10, Th2 cytokine) mRNA expression increased on day 8 in the PBMCs of pregnant cows. Tumor necrosis factor α (TNFα, Th1 cytokine) mRNA expression was stable in all groups. In an in vitro cell culture experiment, IFNT stimulated mRNA expression of ISGs in both PBMCs and PMNs. IFNT stimulated IL-10 mRNA expression in PBMCs, whereas IFNT increased TNFα mRNA levels in PBMCs in vitro. The results suggest that ISGs and IL-10 could be responsive to IFNT before the MRP in peripheral blood immune cells and may be useful target genes for reliable indices of pregnancy before the MRP.