Molecular epidemiology of VapA-positive Rhodococcus equi in thoroughbred horses in Kagoshima,Japan

The prevalence of virulent R. equi having 15- to 17-kDa antigens (VapA) in fecal isolates from 13 thoroughbred foals and their dams on 5 farms in Kagoshima, Japan, and the plasmid profiles of VapA-positive isolates by restriction fragment digestion patterns were investigated to compare the genotypic variation among virulence plasmids of R. equi isolates from Japan. In total, 218 (24.6%) of 886 isolates from the feces of the 13 foals and 13 (12.5%) of 104 isolates from the feces of their dams demonstrated VapA-positive R. equi. Plasmid DNA preparations of 231 virulent isolates from foals and dams were analyzed by restriction enzyme digestion with endonucleases EcoRI, EcoT22I and HindIII and were divided into 3 types: 172 isolates contained a 90-kb type I plasmid, 57 contained a 90-kb type III plasmid and 2 contained a 90-kb type IV plasmid. This study demonstrates a geographic character in the distribution of virulence plasmids found in VapA-positive isolates from thoroughbred foals in Kagoshima.     

Possible roles and functions of LPL1 gene encoding lysophospholipase during early infection by Magnaporthe grisea

The rice blast fungus Magnaporthe grisea differentiates appressoria, which are required to attack its rice plant host. Clone A26, tentatively named LPL1, was previously found to be homologous to the known lysophospholipase genes from our subtractive cDNA library. The LPL1 protein had a consensus motif (GxSxG) and a catalytic triad (S, D, H) of esterases in the deduced amino acid sequence, and the protein expressed in Escherichia coli had lysophospholipase activity. To clarify the functions and possible roles of LPL1, the gene was disrupted by targeted gene replacement. The ΔLPL1 mutants formed fewer appressoria on the hydrophobic surface of GelBond film, and the appressoria had reduced turgor pressure and penetration into cells of the leaf sheath. The ΔLPL1 mutants and wild-type differentiated normal appressoria on other artificial substrata such as polycarbonate plate and on rice leaf sheath. Cytological analysis of the appressoria indicated that ΔLPL1 mutants had a delay in the disappearance of lipid droplets. These findings imply that LPL1, phospholipid metabolism, or both are involved in glycerol biosynthesis and accumulation to generate turgor pressure in the appressorium. LPL1 was, however, dispensable for full pathogenicity, suggesting that other complementary pathways or similar genes related to phospholipid metabolism probably function in M. grisea.     

Deleterious effects of fungi isolated from paddy soils on seminal root of rice

Soil samples were collected from rice paddies at 22 locations in northeastern Honshu, Japan. In 20 of the samples, seedling growth of rice was improved by soil pasteurization (aerated steaming at 60°C for 30 min), although no typical disease symptoms were observed in the seedlings grown in the untreated soil samples. In most locations, rice seedlings grew better in a potting medium containing root material from plants grown in the pasteurized portion of a soil sample than in a medium with root material from plants grown in the unpasteurized portion of the same sample. The results suggest that microorganisms that restrained rice seedling growth may be common in the soils of rice paddies. Approximately 800 isolates were obtained from seedling roots grown in unpasteurized soils and grouped by cultural and microscopic morphologies. The deleterious effects of 79 isolates from 21 major groups, most of which were soil-dwelling taxa, were examined after direct inoculation of the seminal roots. Isolates of Curvularia sp., Cirrenalia sp., Eppicoccum nigrum, Fusarium graminearum, F. oxysporum, Gliocladium virens, Humicola sp., Penicillium sp., Rhizoctonia oryzae-sativae, Sclerotium hydrophilum, Trichoderma aureoviride, and T. harzianum inhibited root growth, suggesting that deleterious root-infecting fungi were more common in paddy soil than previously thought. These fungi may be involved in the restraint of rice seedling growth.     

Effects of leptin on the release of luteinizing hormone, growth hormone and prolactin from cultured bovine anterior pituitary cells

The effects of leptin on the release of luteinizing hormone (LH), growth hormone (GH) and prolactin (PRL) were studied in cultured bovine anterior pituitary (AP) cells in vitro. The AP cells were obtained from fully‐fed Japanese Black steers and were incubated for 3 h with 10^?13 to 10^?7 mol/L of leptin after incubating in Dulbecco's modified Eagle's Medium for 3 days. Leptin significantly increased the concentration of LH in the culture medium by 45 and 44% at doses of 10^?8 and 10^?7 mol/L, respectively, compared with the controls (P < 0.05). Leptin significantly increased the concentration of GH in the culture medium by 14 and 12% at doses of 10^?8 and 10^?7 mol/L, respectively (P < 0.05). Leptin also significantly increased the concentration of PRL in the culture medium by 26% compared with the controls at a dose of 10^?7 mol/L (P < 0.05). These results show that leptin stimulates the release of LH, GH and PRL by acting directly on bovine AP cells from fully‐fed steers.     

Characterization of tetraploid plants regenerated via protoplast culture of Iris fulva and their crossability with Japanese irises

Characteristics such as flower form, size and color of outer and inner perianths, anthocyanins in outer perianths, size, color and fertility of pollen and self-fertility of diploid and tetraploid lines regenerated via protoplast culture of Iris fulva were examined and compared with those of the diploid wild line. Among these characteristics, flower form, inner and outer perianth sizes of the tetraploid lines were noticeable, because these lines had upward flower forms and bigger flowers than diploid lines. Furthermore, reciprocal crosses between diploid or tetraploid lines of I. fulva and I. ensata and those of I. fulva and I. laevigata were performed. Three seedlings were obtained from the cross of tetraploid I. fulva × diploid I. laevigata through embryo rescue. One of them was identified as the interspecific hybrid between tetraploid I. fulva and I. laevigata by flow cytometoric (FCM), cytological and molecular (RAPD) analyses. This is the first report on production of hybrids from these lines. I. fulva has unique brown flowers, and this trait could be very useful for flower color breeding of I. laevigata which lacks this color. Therefore, the hybrid of I. fulva (4×) × I. laevigata may be the best available gene source for brown color breeding of this species.     

The effect of C-2-carboxyphenyl derivatives of certain heterocyclic compounds on root geotropism in cress seedlings

The effects on root geotropism of 2-carboxyphenyl derivatives of various five- and six-membered heterocyclic compounds have been assessed. In an assay involving cress seedlings, appropriately substituted derivatives of isoxazole, 1,3,4- and 1,2,4- oxadiazoles, 1,3,4-thiadiazole, 1,2,4-triazole, thiazoie, pyridine, pyrimidine and pyridazine were highly effective in eliminating the geotropic response.     

Isolation and characterization of a novel potyvirus tentatively named Ornithogalum virus 2

A novel potyvirus, tentatively named Ornithogalum virus 2 (OV-2) because only its nucleotide sequence of the coat protein gene has been revealed, was isolated for the first time from Ornithogalum thyrsoides. OV-2 had a flexuous particle (700–740 nm in length) and was sap and aphid transmissible. The virus had a narrow host range; of 36 test plants in 12 families, only O. thyrsoides and O. dubium were infected. Because the virus caused characteristic stripe mosaic on O. thyrsoides, we propose Ornithogalum stripe mosaic virus (OrSMV), instead of OV-2 for the proper name of the virus. The nucleotide sequence data reported is available in the DDBJ/EMBL/GenBank databases under accession number AB271783.     

Ozonation of a lignin-carbohydrate complex model compound of the benzyl ether type

Evidence for the presence of lignin-carbohydrate bonds of the benzylic ether type in wood and pulps may be obtained by use of ozonation treatment to selectively degrade the lignin. It was found that the benzyl ether bond in 3-(4-hydroxy-3-methoxyphenyl)-2-(2-methoxy-phenoxy)-3-(methyl--d-glucopyranosid-6-O-yl)-1-propanol was rather stable during ozonation in acetic acid-water-methanol 1631 at 0°C. The corresponding acid, 3,4-dihydroxy-2-(methyl--d-glucopyranosid-6-O-yl)-butanoic acid, was found to be the major reaction product. The use of ozonation followed by acid hydrolysis in connection with studies of lignincarbohydrate linkages is briefly discussed.Part of this paper was presented at the 10th ISWPC, Yokohama, 1999     

Changes in spatial patterns of sika deer distribution and herbivory of planted seedlings: a comparison before and after deer population control by culling

Sika deer (Cervus nippon) sometimes cause extensive damage to planted tree seedlings. To evaluate the effects of culling on the spatial distribution patterns of sika deer and browse damage to planted seedlings, we compared the data collected before and after experimental culling in a cool, temperate, mixed forest on Kyushu Island, Japan. Experimental culling, conducted in an area of 1 km², removed five, four and two sika deer in April, June and October 2011, respectively. During the year before culling, the spatial pattern of the number of sika deer caught on camera corresponded to the predicted sika deer density. Sika deer immediately browsed planted seedlings after the initial planting. The cumulative number of browsed seedlings increased over time, especially in winter. The spatial pattern of the cumulative number of sika deer caught on camera corresponded to that of browsed seedlings at the year’s end. During the year when culling was conducted, the number of sika deer caught on camera decreased around the center of the study site where the culling was conducted and the number of browsed seedlings decreased. During the year following culling, the cumulative number of browsed seedlings was very similar to that in the year before the culling, while the same low number of sika deer was caught on camera. These results indicate that the effects of deer culling resulted in decreased levels of sika deer appearance and browse damage for more than 1 year and for several months, respectively.