Cloning of the pathogenicity-related gene <Emphasis Type="Italic">FPD1</Emphasis> in <Emphasis Type="Italic">Fusarium oxysporum</Emphasis> f. sp. <Emphasis Type="Italic">lycopersici</Emphasis>

We selected a reduced-pathogenicity mutant of Fusarium oxysporum f. sp. lycopersici, a tomato wilt pathogen, from the transformants generated by restriction enzyme-mediated integration (REMI) transformation. The gene tagged with the plasmid in the mutant was predicted to encode a protein of 321 amino acids and was designated FPD1. Homology search showed its partial similarity to a chloride conductance regulatory protein of Xenopus, suggesting that FPD1 is a transmembrane protein. Although the function of FPD1 has not been identified, it does participate in the pathogenicity of F. oxysporum f. sp. lycopersici because FPD1-deficient mutants reproduced the reduced pathogenicity on tomato.The nucleotide sequence data reported are available in the DDBJ/EMBL/GenBank databases under the accession number AB110097     

Therapeutic and prophylactic efficacy of milbemycin oxime (Interceptor) against Thelazia callipaeda in naturally exposed dogs

Two trials were conducted to evaluate the therapeutic and prophylactic activity of milbemycin oxime (Interceptor, Novartis Animal Health) against the eye-worm Thelazia callipaeda (Spirurida, Thelaziidae) infection. In Trial 1, the therapeutic efficacy of milbemycin oxime was evaluated in 55 naturally infected dogs treated with min. 0.5mg/kg milbemycin oxime. The dogs were clinically examined for the presence of eyeworms before and again 7 days after treatment. Dogs still positive were given a second treatment and re-checked again a week later. Forty-eight of the 55 dogs tested negative 1 week after treatment (87.3% reduction of infection rate). Following the second treatment 6 of these 7 dogs tested negative 1 week later resulting, after two treatments, in a reduction of infection rate of 98.2%. In Trial 2, the prophylactic efficacy of milbemycin oxime was evaluated in 60 uninfected dogs. Thirty dogs were treated with milbemycin oxime monthly from June to November with the recommended dose rate for the prevention of heartworm disease (> or =0.5mg/kg), 30 dogs served as untreated controls. At the end of the trail 1 dog in the treated group and 10 dogs in the control group became infected during the trial. The incidence of infection differed significantly between treated and control dogs (p=0.0056). The efficacy of the prophylactic use of a monthly treatment with milbemycin oxime showed 90% efficacy in reducing T. callipaeda infection rate.     

Effects of the lateral growth rate on wood quality parameters of Eucalyptus grandis from different latitudes in Brazil and Argentina

Climate change resulting from increased atmospheric carbon dioxide (CO₂) and shortages of fossil fuels such as petroleum are major problems worldwide. Under these conditions, demand for woody biomass resources is increasing. We investigated the feasibility of using fast-growing Eucalyptus grandis for material production. Samples of E. grandis were collected from four plantations in different latitude divisions, including tropical and subtropical Brazil and subtropical Argentina. Various xylem qualities were measured and related to the lateral growth rate. Lateral growth rate did not significantly affect the longitudinal released strain of the surface growth stresses or the xylem density at any of the sampling sites. Higher lateral growth rate, higher values of xylem density, and lower absolute values of the released strain were observed in plantations closer to the equator. Higher growth rates in tropical climate promote longer fiber length. In subtropical plantations, smaller diameter trees will produce tension wood with smaller microfibril angles. Planting E. grandis closer to the equator thus produces higher quality wood than in plantations at lower latitudes.     

Generation process of growth stresses in cell walls: Relation between longitudinal released strain and chemical composition

Summary To advance the discussion on the evolution mechanism of tree growth stresses, the relation between released strain and the chemical components was investigated experimentally. The expansive released strain in the longitudinal direction assumed large values as the lignin content increased in the compression wood, but there was no relation between released strain and lignin content in the normal wood region. The contractive released strain assumed large values as the cellulose content and its crystallinity increased. Their correlation was very high and clear. From these facts, it is considered that the lignin deposition plays an important role in the generation of the growth stresses in compression wood but is not important in normal or tension wood regions. Cellulose microfibrils contract along their longitudinal axis during cell maturation, and the stress included by the contraction creates a longitudinal growth stress in normal and tension woods.The authors thank Prof. R. R. Archer, University of Massachusetts, for his valuable advice. A part of this research was supported by a grant under the Monbusho International Scientific Research Program     

Direct detection of equine herpesvirus type 1 DNA in nasal swabs by loop-mediated isothermal amplification (LAMP)

We evaluated loop-mediated isothermal amplification (LAMP) as a means of detecting equine herpesvirus type 1 (EHV-1) DNA directly from nasal swabs. To increase the sensitivity, we added a step in which the samples were heat-treated to the original LAMP procedure. The detection limit of the LAMP assay with heat treatment was 10 times more sensitive than the original LAMP assay even when the DNA extraction step was omitted. In addition, the LAMP assay with heat treatment was more sensitive than the original LAMP assay and the polymerase chain reaction using clinical samples. The LAMP assay with heat treatment is easy to perform and so should be applicable to the diagnosis of EHV-1 infections in clinical laboratories.     

Isolation and Structure Elucidation of New Cytotoxic Macrolides Halosmysins B and C from the Fungus Halosphaeriaceae sp. Associated with a Marine Alga

Two new cytotoxic metabolites, halosmysins B and C, have been isolated from the fungus Halosphaeriaceae sp. OUPS-135D-4 separated from the marine alga Sargassum thunbergii. These chemical structures have been elucidated by 1D and 2D NMR, and HRFABMS spectral analyses. The new compounds had the same 14-membered macrodiolide skeleton as halosmysin A, which was isolated from this fungal strain previously. As the unique structural feature, a diketopiperazine derivative and a sugar are conjugated to the 14-membered ring of halosmysins B and C, respectively. The absolute stereostructures of them were elucidated by the chemical derivatization such as a hydrolysis, the comparison with the known compounds (6R,11R,12R,14R)-colletodiol and halosmysin A, and a HPLC analysis of sugar. In addition, their cytotoxicities were assessed using murine P388 leukemia, human HL-60 leukemia, and murine L1210 leukemia cell lines. Halosmysin B was shown to be potent against all of them, with IC₅₀ values ranging from 8.2 ± 1.8 to 20.5 ± 3.6 μM, though these values were slightly higher than those of halosmysin A.     

Comparison of Iron Availability in Leaves of Barley and Rice

Iron (Fe) is an essential trace element in all eukaryotes. In higher plants, Fe deficiency causes interveinal chlorosis in young leaves. However, in barley and rice, both of which are "Strategy II" plants, the degree and the pattern of Fe-deficiency symptoms differ. In the present study, barley and rice plants were grown in the same container, i.e., by "coculturing," to compensate for the amount of mugineic acids in rice in the nutrient solution. We examined the differential availability of Fe for distribution and retranslocation in shoots between barley and rice without considering the difference in the iron acquisition ability, which is affected by the differential mugineic acid secretion between barley and rice. Although the Fe concentration of young barley leaves had decreased under the coculture conditions, the SPAD value was similar to that in monocultured barley. In contrast, although there was an increase in the Fe concentration of the young leaves of cocultured rice, the SPAD value decreased, as in the case of monocultured rice. Rice accumulated Fe in old leaves, whereas in barley Fe was efficiently distributed to young leaves. Therefore, the SPAD value of the second leaf in rice remained constantly high. The Fe concentration of the second leaf in barley decreased under Fe-deficient coculture conditions, the SPAD value decreased and the senescence of the second leaf become accelerated. ⁵⁹Fe pulse-labeling experiments suggested that in barley Fe was more efficiently retranslocated from old leaves to young leaves than that in rice. As a result, the level of Fe present in the fraction with a molecular weight lower than the 10,000/water-soluble Fe ratio was higher in the old leaves of barley than in the old leaves of rice under Fe-deficient conditions. Based on the results obtained, we suggest that the distribution and retranslocation characteristics of internal Fe in barley may be well adapted to Fe deficiency.     

The Kidneys of Infant Mice are not Sensitive to the Food Mycotoxin Contaminant Nivalenol

Nivalenol (NIV) is a trichothecene mycotoxin produced by Fusarium fungi that frequently contaminates agricultural commodities. Dietary administration of NIV to adult mice affects the renal glomeruli, but data about NIV toxicity in human infants are limited. To evaluate the effects of NIV on infant kidneys, 3-week-old male ICR-derived glomerulonephritis (ICGN) and ICR mice were administered 0, 4, 8 or 16 ppm NIV in diet for 4 weeks, and their renal status was compared with age-matched or adult ICR mice. In ICGN mice, the number of glomeruli showing mesangial expansion and α-smooth muscle actin (SMA)-positive mesangial cells was higher with 16 ppm NIV compared with controls. No other significant differences were observed in ICGN mice. In infant ICR mice, the IgA serum concentrations were significantly elevated without glomerular morphological changes in the 16 ppm NIV group. There was no difference in NIV sensitivity in the kidneys of infant ICGN and ICR mice. These data suggest that the kidneys in infant mice are not sensitive to nivalenol under the present conditions.     

Studies on inter-relationship of various ions in absorption by tobacco plants IV

The foliar symptoms of magnesium deficiency occur not only by the decrease in the contents of exchangeable magnesium in soil, but the antagonistic interaction among the nutrient elements in absorption. The relationship between phosphorus and magnesium in absorption was reported previously (1). In this paper influence of potassium and calcium on the magnesium absorption of tobacco plant was investigated.     

Beta 3-adrenergic agonist up-regulates uncoupling proteins 2 and 3 in skeletal muscle of the mouse

Chronic stimulation of the beta3-adrenergic receptor (AR) in obese animals resulted in a reduced adiposity associated with an increased expression of thermogenic uncoupling protein (UCP)1 in adipose tissues. In this study, the mRNA expression of newly cloned UCP isoforms (UCP2 and UCP3) were examined in obese yellow KK and C57BL control mice. UCP2 mRNA was found in all tissues examined, with higher levels in adipose tissues and skeletal muscle of the obese mice. UCP3 mRNA was expressed in skeletal muscle, heart and brown adipose tissue similarly in the two mouse strains. Daily injection of a selective beta3-adrenergic agonist, CL316,243 (0.1 mg/kg), for 10 days resulted in a marked reduction of white fat pad weight and 1.8-4.8-fold increase in the mRNA levels of UCP2 and UCP3 in skeletal muscle of obese mice. No noticeable change in the UCP2 and 3 mRNA levels was found in brown and white adipose tissues. It was also found that CL316,243 injection produced a marked and sustained elevation of the plasma free fatty acid level. These results, together with our previous findings of the fatty acid-induced UCP expression in a myocyte cell line in vitro, suggest that the beta3-AR agonist-induced UCP expression in skeletal muscle may be mediated through the elevated plasma free fatty acids. It was also suggested that anti-obesity effect of beta3-AR agonists is attributable to increased thermogenesis not only by UCP1 but also by UCP2 and UCP3.