Virological studies of paramyxovirus type 1 infection of pigeons

Over a period of three months in 1985, paramyxovirus type 1 infection was demonstrated for the first time in Canada, in six flocks of pigeons in Ontario, Alberta, and British Columbia. The paramyxovirus type 1 isolates did not cause clinical disease when serially passaged four times in four-to six-week-old chickens, and isolates were classified as lentogenic before and after such serial passage. Further cases of paramyxovirus type 1 clinical disease have not been reported since the last of these six outbreaks in August 1985.     

Transfer of the Rsv3 locus from ‘Harosoy’ for resistance to soybean mosaic virus strains C and D in Japan

Resistance to soybean mosaic virus (SMV) is imperative for soybean (Glycine max (L.) Merr.) production in the Tohoku region. Molecular markers for SMV resistance were previously reported for U.S. SMV strains, but they cannot be applied because of the differences in strain classification between Japan and the U.S. A U.S. variety ‘Harosoy’ has been used mainly as a donor of resistance to SMV strains C and D in a Japanese breeding program, resulting in resistant varieties such as ‘Fukuibuki.’ Because ‘Harosoy’ harbors the Rsv3 gene conferring resistance to the virulent SMV strain groups, G5 through G7, it appears that the Rsv3 gene confers resistance to strains C and D. In this study, we introduced resistance to the two strains from ‘Fukuibuki’ into a leading variety ‘Ohsuzu’ by recurrent backcrossing with marker-assisted selection. All lines selected with markers near Rsv3 showed resistance to the strains, suggesting that the Rsv3 locus is responsible for the resistance. Three years of trials showed that one of the breeding lines, ‘Tohoku 169,’ was equivalent to ‘Ohsuzu’ with respect to agricultural characteristics such as seed size, maturity date, and seed yield, except for the SMV resistance.     

Immunohistochemical localization of enzymes related to lignin biosynthesis in the primary xylem of hybrid aspen

We have investigated the spatial regulation of the accumulation of enzymes involved in the biosynthesis of shikimate and lignin during differentiation of primary xylem from the apical meristem via procambium in hybrid aspen (Populus sieboldii x Populus grandidentata). Immuohistochemical staining revealed that, in the top part of shoots, lignification began in a single or just a few adjacent vessel elements and subsequently spread to neighboring cells. The spatial localization of 3-deoxy-D-arabino-heptulosonate 7-phosphate synthase (DAHPS), which is one of the key enzymes in the shikimate pathway, was tightly correlated with the cell-specific deposition of lignin in the primary xylem. We also found that the spatial localization of enzymes in the general phenylpropanoid pathway and in the lignin-specific pathway was closely associated with the cell-specific deposition of lignin and the accumulation of DAHPS. Our data suggest that enzymes that act in the shikimate, general phenylpropanoid, and lignin-specific pathways are initially produced and function coordinately in a single or a few adjacent elements at the start of primary xylem development.     

Growth response to the stress of low osmotic potential,salinity and high pH in cultured shoot of Chinese poplars

In vitro cultured shoots of three Chinese poplar species (Populus tomentosa, P. alba cv. Pyramidalis andP. euphratica) and an improved poplar (FS-51), as a control, were exposed to the stress of low osmotic potential, salinity and high pH, and the shoot growth was measured.P. tomentosa andP. euphratica could grow even under the high osmotic stress (0.5m mannitol; −1.7 MPa).P. euphratica shoots which were previously cultured on the medium containing 0.7m mannitol (−2.4 MPa) recovered its growth after being transplanted to the medium without mannitol. The shoot growth ofP. euphratica was not so reduced even on the medium containing 100 mM NaCl. However, growth of the other poplars markedly reduced with 10 mM NaCl. Medium pH (pH 5.8–10) also did not affect the shoot growth ofP. euphratica. These poplars have different tolerance to the stresses in cultured shootin vitro. P. euphratica was extremely tolerant to the stresses which are possible to damage plants in dry land. A part of this study was presented at the 104th Annual Meeting of the Japanese Forestry Society. (1993).     

Freezing behaviors in leaf buds of cold-hardy conifers visualized by NMR microscopy

1H-Nuclear magnetic resonance (NMR) microscopy was used to study freezing behavior in wintering leaf buds of Momi fir (Abies firma Sieb. et Zucc.) and Japanese red pine (Pinus densiflora Sieb. et Zucc.). The images acquired predominantly reflected the density of mobile (i.e., non-ice) protons mainly from unfrozen water. By comparing images taken at various subfreezing temperatures, we determined which tissues produced the high and low temperature exotherms detected by differential thermal analyses. Typical extra-organ freezing was successfully imaged in leaf buds of A. firma. The bud scales readily froze at -7 degrees C, but shoot primordia remained supercooled to -14 degrees C in December buds and to -21 degrees C in March buds. The size of supercooled shoot primordia was reduced with decreasing temperature, indicating a gradual decrease in water content of the shoot primordia. In contrast, the signal from shoot primordia of P. densiflora disappeared between -7 and -14 degrees C, corresponding to the high temperature exotherm at -8 degrees C, indicating extracellular freezing of the shoot primordia. The xylem and bark tissues readily froze at -7 degrees C in A. firma and between -7 and -14 degrees C in P. densiflora. We conclude that NMR microscopy can noninvasively provide more spatially specific information about freezing behavior in leaf buds than traditional methods such as differential thermal analysis. In particular, it allows the organized and harmonized freezing behaviors in complex organs to be visualized directly thereby revealing the diversity of mechanisms involved in freezing behaviors.     

Technique for screening immune-enhancing polysaccharides in food using 1,25-dihydroxyvitamin D3-differentiated HL60 cells

A technique for screening immune-enhancing polysaccharides in food using the phagocytotic activity of 1,25-dihydroxyvitamin D3 (VD3)-differentiated HL60 cells is presented. HL60 cells, a human acute promyelocytic cell line, can differentiate along the monocytic lineage following exposure to VD3 or phorbol-12-myristate-13-acetate (PMA). For differentiated cells along the monocytic pathway, HL60 cells were maintained in RPMI 1640 medium supplemented with 10% FBS and 120 nM VD3 for more than 1 week. VD3-differentiated HL60 cells were seeded into 48-well plates, YG-labeled microspheres and polysaccharides were added and mixed using a plate shaker at 1100 rpm for 30 s, and then the mixture was incubated overnight at 37 degrees C in 5% CO2. The cells were fixed with 2% formaldehyde and resuspended in phosphate-buffered saline. The rate of phagocytosis was measured with a flow cytometer. VD3-differentiated cells but not non- and PMA-differentiated cells resulted in an elevation of phagocytic activity by various immune-enhancing polysaccharides in foods.     

2'-epi-orobanchol and solanacol, two unique strigolactones, germination stimulants for root parasitic weeds, produced by tobacco

Germination stimulants for root holoparasitic weeds broomrapes ( Orobanche and Phelipanche spp.) produced by tobacco ( Nicotiana tabacum L.) were purified and characterized. The root exudates of tobacco contained at least five different stimulants, and LC-MS/MS analyses revealed that four of them were strigolactones; a tetradehydrostrigol isomer, a didehydrostrigol isomer, and two strigol isomers. The two isomers of strigol were identified as (+)-orobanchol and its 2'-epimer by comparison of NMR and GC- and LC-MS data with those of synthetic standards. The structure of the tetradehydrostrigol isomer, the major stimulant of the bright yellow tobacco cultivars, was determined as 4-alpha-hydroxy-5,8-dimethyl-GR24 [( E)-4-alpha-hydroxy-5,8-dimethyl-3-(4-methyl-5-oxo-2,5-dihydrofuran-2-yloxy)methylene)-3a,4-dihydro-3 H-indeno[1,2- b]furan-2(8b H)-one] and named solanacol. 2'-Epi-orobanchol and solanacol are the first natural strigolactones having a 2'-epi stereochemistry and a benzene ring, respectively.     

Systemic candidiasis in a dog, developing spondylitis

A 4-year-old male Shiba dog initially presented with pain of an undetermined origin and hypersensitivity to touch. Seven days later, the dog developed ataxia, hind-leg weakness and knuckling. The dog died on 20 days after presentation. Postmortem examination revealed a mass in the body of thoracic vertebra. Histopathologically, the mass consisted of granulomatous inflammation, including fungal organisms that were immunohistochemically positive for Candida albicans. Similar granulomatous lesions were observed in the systemic lymph nodes, kidneys, pancreas, spleen, prostate gland, thyroid glands and heart. This case was diagnosed as systemic candidiasis with spondylitis.     

Establishment and characterization of a cell line, MCO-Y4, derived from canine mammary gland osteosarcoma

A cell line, MCO-Y4, was established from a mammary gland osteosarcoma of a 16-year-old female mongrel dog. Histopathologically the tumor was composed of osteoblastic cells with an osteoid meshwork and chondroid matrix. The mean doubling time of the cells at the 93rd passage was 32.39+/-4.66 hr. Immunohistochemically, the osteoblastic and chondroblastic cells were positive for bone morphogenetic protein (BMP)-2/4 and BMP receptor (BMPR) II. The cultured cells were spindle in shape during the growth and the confluent phases. No tumor matrix was detected in the culture dish by alcian blue staining or von-Kossa silver impregnation. MCO-Y4 cells on the chamber slides showed intense immunoreactivity for BMP-2/4 and BMPR II. Noggin, an antagonist for BMP-2/4, showed the growth inhibition on MCO-Y4 cells. In addition, fibronectin might be potential for stimulating growth of MCO-Y4 cells. When transplanted into severe combined immunodeficiency mice, the cells formed tumors consisting of solid proliferation of osteoblastic and fibroblastic cells with woven-bone trabeculae. These tumor cells were intensely positive for BMP-2/4 and BMPR II. Our results suggested that the cell line might be useful for studying the role of BMPs in canine osteosarcoma and the mechanism of ossification.