Development of a rapid method for identifying asparagus super-males using, N-(4-chloro-2-trifluoromethylphenyl)-N'- propoxyacetamidine to induce flowering

In order to shorten the term for the identification of asparagus super-maleplants, a method usingN-(4-chloro-2-trifluoromethylphenyl)-N'-pro-poxyacetamidine (AM12) toinduce flowering was developed. This method is conducted as follows: seedsfrom andromonoecious flowers are treated with AM12 to induce flowersto form on the seedlings, the new male flowers are crossed with normalfemale flowers, and the progeny seeds are again treated with AM12 toidentify super-male plants from the sex ratio of the seedlings.Asparagus officinalis cv. `UC157' seeds were treated with AM12.The seedlings were induced to flower at a high frequency in 25 days. Thesex ratio of the plants was 1:1 and the male flowers had the pollengermination ability. One male flower induced by AM12 was crossed withfemale plant. This cross produced many progeny seeds. The seeds weretreated again with AM12, and induced to flower. Two super-male plantswere found among the progeny of andromonoecious flowers of the all-malecultivar `Gijnlim' within six months by this method. This method thusshortens the time for identifying super-males. Since the female flowers werefertile as well, AM12 treatment would also be effective for cross breeding.     

Species-specific PCRs differentiate <Emphasis Type="Italic">Rosellinia necatrix</Emphasis> from <Emphasis Type="Italic">R. compacta</Emphasis> as the prevalent cause of white root rot in Japan

Rosellinia compacta, described recently, resembles R. necatrix and also causes white root rot. Here a species-specific PCR was developed for R. compacta, and the two R. necatrix-specific primer sets already available were validated in terms of species specificity. PCRs using the primer sets for R. necatrix amplified specific products exclusively from R. necatrix isolates. The R. compacta-specific primer set exclusively detected R. compacta, which appears to be a rare but widely distributed species. We conclude that R. necatrix is the major cause of the disease in Japan but that the involvement of R. compacta should be studied further.     

Inhibition of chitin biosynthesis by buprofezin analogs in relation to their activity controlling Nilaparvata lugens Stål

Buprofezin (Applaud, 2-tert-butylimino-3-isopropyl-5-phenyl-3,4,5,6-tetrahydro-2H-1,3,5-thiadiazin-4-one) strongly inhibited the [³H]chitin synthesis from N-acetyl-d-[1-³H]glucosamine in the brown rice planthopper, Nilaparvata lugens Stål. No inhibition was observed for [³H]-labeled protein biosynthesis from [5-³H]glucose or l-[3,5-³H]tyrosine but [³H]-labeled nucleic acid synthesis from [5-³H]glucose was weakly reduced by buprofezin. The lethal activity of buprofezin analogs related well to their inhibitory potency against chitin biosynthesis in N. lugens nymphs.     

Economic importance of theileriosis in Japan

Tropical Animal Health and Production -     

Application of renal microangiography to normal and diseased kidneys of cattle and mice

Use of microangiography is now essential for the study of microcirculation in various organs. Renal microangiographic studies have been reported in rats, rabbits, dogs, human beings, and mice. However, we could not find any report on use of the technique in cattle, despite high incidence of renal disease in that species. The perfusion technique used in mice was improved over that of our previous report, and was applied to normal and diseased bovine kidneys. For the microangiographic technique, composition of the contrast medium, pressure of the injection, duration of perfusion, and washing of kidneys with heparinized saline solution before perfusion are important. In cattle, 1- to 2-mm-thick sections of the kidneys were generally necessary to observe renal vasculature: arcuate and interlobular arteries, afferent arterioles, and glomerular capillaries. In normal bovine kidneys, the angiographic and microangiographic findings were easily recognized as normal, compared with those of normal mice. In affected bovine kidneys, which histologically represented glomerulonephritis and pyelonephritis, angiography and microangiography revealed corresponding findings.     

Vertical transmission of bovine leukemia virus and bovine immunodeficiency virus in dairy cattle herds.

Vertical transmission of bovine leukemia virus (BLV) and bovine immunodeficiency virus (BIV) was investigated in five dairy cattle herds in Hokkaido, where 36.1 and 17.0% of cattle were BLV and BIV seropositive, respectively, and 9.9% of dams were co-infected with both BIV and BLV. Twenty six cases of offspring born from dams infected with only BLV (17 cases) or with both BIV and BLV (9 cases) were examined for the presence of BLV and BIV before and after colostrum feeding by polymerase chain reaction (PCR) and syncytium assay. After birth, all calves were separated immediately from their dams. The offspring born from BLV-positive dams were BLV-negative before colostrum feeding, suggesting that no transplacental transmission had occurred. Thereafter, these offspring were fed colostrum or milk from their dams, but still remained BLV-negative. The other offspring born from BLV-positive dams were fed with BLV-negative colostrum, or with pasteurized BLV-positive colostrum. All these calves remained negative for BLV infection, suggesting that in utero transmission of BLV is negligible. In the case of offspring born from dams co-infected with BLV and BIV, calves were BIV-positive before colostrum feeding at 1 day after the birth, indicating in utero transmission of BIV. After colostrum feeding from their dams, newborn calves became BLV-positive. In addition, one calf was BLV-positive even before colostrum feeding. These results suggest that BIV can be transmitted to offspring in utero, and that BLV can be transmitted through colostrum or milk if dams are infected with both BIV and BLV.     

Comparison of estrogen responsive genes in the mouse uterus, vagina and mammary gland

Female reproductive organs are mainly regulated by estrogen and progesterone. Specifically, the uterus, vagina and mammary gland show organ-specific mitosis and morphological changes during proliferative events, such as estrous cycle, gestation and lactation. The mechanism underlying these organ-specific estrogen-dependent events is still unknown. We examined, therefore, global gene expression in the mature uterus, vagina and mammary gland of ovariectomized adult mice 6 hr after an injection of 5 microg/kg 17beta-estradiol (E2) using a microarray method in order to identify primary E2-responsive genes. Half of the E2 up-regulated genes in the uterus were similar to those in the vagina. E2 up-regulated the expression of Insulin-like growth factor 1 (Igf-1) genes in the uterus and vagina. In the vagina, E2 up-regulated the expression of IGF binding proteins (Igfbp2 and Igfbp5). In the mammary gland, unlike the uterus and vagina, no gene showed altered expression 6 hr after the E2 exposure. These results suggest that expression of Igf-1 and morphogenesis genes is regulated by E2 in an organ-specific manner, and it is supported by the results of BrdU labeling showing E2-induced mitosis in the uterus and vagina except the mammary gland. The differences in organ specificity in response to E2 may be attributed by differences in gene expression regulated by E2 in female reproductive organs. The candidate estrogen-responsive genes in the uterus and vagina identified by profiling provide an important foundation understanding functional mechanisms of estrogen regulating morphogenesis and maintenance of each reproductive organ.     

Molecular characterization of a myosin alkali light chain-like protein, a "concealed" antigen from the hard tick Haemaphysalis qinghaiensis

There should be some differences between antibodies generated by feeding ticks on animals and those derived by immunizing animals with tick extracts. Here, we found serum collected from sheep immunized with Haemaphysalis qinghaiensis salivary gland extracts could detect two more protein bands with molecular weights of 22 and 37 kDa (P22 and P37) on Western blots of extracts of tick salivary glands than serum from tick infected animals. Rabbit anti-H. qinghaiensis differential protein immune serum was then generated from P22 and P37 and was used to immunoscreen a cDNA library constructed from salivary glands, Malpighian tubules and ovaries of partially engorged H. qinghaiensis. A cDNA contains an open reading frame of 483 bp that codes for 160 amino acid residues with a coding capacity of 18 kDa was cloned and designated Hq02. Expression analysis by RT-PCR showed that this gene is expressed in salivary glands, midguts, other organs and different developmental stages of H. qinghaiensis. The predicted amino acid sequence of the Hq02 gene had high homology to some known myosin alkali light chain (MLC) proteins. A fusion protein consisting of 130 amino acids of Hq02 protein and 335 amino acids of T7 gene 10 protein was expressed in Escherichia coli and used to immunize sheep. Western blot showed that only rabbit anti-H. qinghaiensis differential protein immune serum could recognize the expressed Hq02 protein, while rabbit anti-H. qinghaiensis saliva immune could not. This proved Hq02 protein was a "concealed" antigen. Immunization with the recombinant Hq02 conferred a 21.8% reduction of engorgement weight for adult female ticks that fed on the immunized sheep. This is the first report of tick myosin alkali light chain and the function of this protein is discussed.     

Changes in the aroma compounds of sake during aging

Changes in the aroma of sake during aging were investigated by aroma extract dilution analysis (AEDA) and quantitative analysis using the stir bar sorptive extraction method. In AEDA, more odor zones were detected in aged sake than in fresh sake. The dilution factors of aldehydes, polysulfides, and some esters were greater in the aged sake, and their increase during aging was confirmed through a quantitative analysis of sake stored for 0-35 years. Among these compounds, 3-methylbutanal, methional, and dimethyltrisulfide (DMTS) were present in aged sake at concentrations exceeding their odor thresholds, and the highest odor active value was observed for DMTS. Sensory tests showed that supplementation with DMTS contributed to both the total odor intensity and the sulfury odor of aged sake aroma.