QTLs in barley controlling seedling elongation of deep-sown seeds

In drought areas, in which topsoil moisture is low, barley seeds are generally sown in the subsoil (deep-seeding). In order for the shoots of germinating seeds to emerge from the deep soil cover, the coleoptile and the first internode of the seedlings must elongate as an adaptive response to deep-seeding. Here, we have mapped quantitative trait loci (QTLs) for these adaptive characteristics. Elongation of the coleoptile and first internode was investigated using seeds sown under two soil cover conditions: at a depth of 9 cm beneath a soil mixture; and, at a depth of 12 cm beneath vermiculite. We identified multiple alleles for increased coleoptile and first internode elongation using a doubled haploid population of 150 lines generated from a cross between the barley cultivars Harrington and TR306. Composite interval mapping analyses of the data revealed two moderate and eleven small effect QTLs, with at least one QTL on each chromosome. The QTLs on chromosomes 5H and 7H had moderate effects on coleoptile elongation (18.5–27.6% of PVE: phenotypic variance explained; 2.6–3.2 mm of Add: additive effect) and first internode elongation (PVE: 16.6–19.6%; Add: 3.1–3.2 mm). The small effect QTLs showed PVEs of less than 15% and an Add range of 1.2–3.2 mm for both characters. A marker assisted selection approach, using markers linked to the QTLs for seedling elongation at deep-seeding, may eventually enable development of drought tolerant barley hybrids.     

Effects on flowering and seed yield of dominant alleles at maturity loci E2 and E3 in a Japanese cultivar,Enrei

‘Enrei’ is the second leading variety of soybean (Glycine max (L.) Merr.) in Japan. Its cultivation area is mainly restricted to the Hokuriku region. In order to expand the adaptability of ‘Enrei’, we developed two near-isogenic lines (NILs) of ‘Enrei’ for the dominant alleles controlling late flowering at the maturity loci, E2 and E3, by backcrossing with marker-assisted selection. The resultant NILs and the original variety were evaluated for flowering, maturity, seed productivity and other agronomic traits in five different locations. Expectedly, NILs with E2 or E3 alleles flowered later than the original variety in most locations. These NILs produced comparatively larger plants in all locations. Seed yields were improved by E2 and E3 in the southern location or in late-sowing conditions, whereas the NIL for E2 exhibited almost the same or lower productivity in the northern locations due to higher degrees of lodging. Seed quality-related traits, such as 100-seed weight and protein content, were not significantly different between the original variety and its NILs. These results suggest that the modification of genotypes at maturity loci provides new varieties that are adaptive to environments of different latitudes while retaining almost the same seed quality as that of the original.     

Morphology and size of embryos recovered from superovulated cows.

A total of 691 normal embryos were recovered from 183 superovulated donor cows on the 5th and 6th days after the first insemination, and were examined for their morphology and size in relation to their developmental stage. There was no significant difference in the thickness of the zona pellucida, the diameter of the cell mass, and the overall diameter of the embryos among zygotes, 2-, 4-, 8- and 16-cell embryos, and morulae. In the blastocyst stage, however, the diameter of the cell mass and the overall embryo diameter were significantly greater and the zona pellucida was significantly thinner than in the earlier-stage embryos. The volume of the blastomere significantly decreased from zygote to morula in proportion to the increase in the number of blastomeres. The volume of the cell mass of 2-cell embryos was decreased by about 30% compared with that of zygotes and no increase in the volume of the cell mass was observed during the progression from 2-cell stage to morula. The diameter of the cell mass and the overall diameter of morulae recovered on the 6th day after the first insemination were significantly greater than those of morulae recovered on the 5th day.     

Physicochemical properties of akabane virus: A member of the simbu arbovirus group of the family bunyaviridae

The multiplication of Akabane virus was not inhibited in the presence of 5-iodo-2′-deoxyuridine, indicating the presence of RNA. The virus was considered to have an envelope, as it was sensitive to ether and chloroform. It was readily inactivated by deoxycholate and trypsin, but was not precipitated by protamine sulphate. The virus was very labile at pH 3 and also rather heat-labile. Akabane virus was readily filtered through membrane filters of 200 or 100-nm pore size, but not through 50-nm filters. Equilibrium centrifugation in a CsCl density gradient gave a peak of infectivity and hemagglutinin at a density of 1.22 g/ml. The peak fractions thus obtained contained numerous virus particles, roughly spherical, variable in size, 70 to 130 nm in diameter, and mostly having a ragged, closely adherent envelope with projections, when examined, following phosphotungstic acid negative staining, in an electron microscope.     

Bacterial black spot caused by <Emphasis Type="Italic">Burkholderia andropogonis</Emphasis> on <Emphasis Type="Italic">Odontoglossum</Emphasis> and intergeneric hybrid orchids

A new bacterial black spot disease was observed on Odontoglossum, Odontioda, Odontocidium, and Vuylstekeara orchids in Japan. Typical symptoms on the leaves were dark or black spots (or both) with a yellow halo. The causal agent was identified as Burkholderia andropogonis (Smith 1911) Gillis, Van Van, Bardin, Goor, Hebbar, Willems, Segers, Kersters, Heulin and Fernandez 1995. The isolates were pathogenic on four original host orchids, Phalaenopsis orchid, and tulip; they were not pathogenic on white clover or corn after needle stab inoculation. An antibiotic bactericide (oxytetracycline/streptomycin mixture WP) was most effective for controlling the disease.     

Isolation and monolayer culture of bovine oviduct epithelial cells

Oviduct epithelia obtained from 32 cows were cultured. The oviducts were classified into follicular and luteal phases and divided into ampulla and isthmus regions. The epithelial cells were dissociated by enzyme digestion and cultured in plastic dishes with Dulbecco's modified Eagle's medium/Ham's F12 (1:1) containing 10% calf serum. After enzyme treatment, the epithelial suspension showed free ciliated and non-ciliated cells, and cell mass. The non-ciliated cells contained secretory granules in the cytoplasm. The cell mass was composed of ciliated and secretory cells. The cell mass adhered to the dish within 12-24 hr, while the free ciliated cells attached on Day 2 of the culture. The cells grew into confluent monolayers on Day 4. The cell monolayers contained ciliated and non-ciliated cells. The monolayered non-ciliated cells showed a few secretory granules. When the cells were further cultured without subculturing, ciliary activity diminished on Day 5 and was rarely detected on Day 9. When the cells were subcultured on Day 3, ciliary movement was detected on the monolayers for only 2 days. Cell mass that did not adhere to the dish and remained floating in the medium formed ball-like structures on Day 2. Active ciliary beating was observed on the cells that were cultured in the medium supplemented with 10(-5) and 10(-9) M estradiol-17 beta, however, the ciliary activity diminished on Day 5. No difference in the cell growth was observed between the follicular and luteal phases or between the ampulla and isthmus regions.     

Hypereosinophilic syndrome in two cats

Two cats showing chronic vomiting, diarrhea and weight loss were found to have leukocytosis with marked eosinophilia. Both cats were diagnosed with hypereosinophilic syndrome by the findings of increased eosinophils and their precursors in the bone marrow, eosinophilic infiltration into multiple organs, and exclusion of other causes for eosinophilia. Although cytoreductive chemotherapy with hydroxycarbamide and prednisolone was performed, these two cats died 48 days and 91 days after the initial presentation.     

Injection and sampling methods for drug residue study in calf muscle

Eight calves, weighing 50-150 kg, were given intramuscularly 5 ml of ampicillin (ABPC) aqueous suspensions (200 mg potency/ml) in their right and left gluteal and femoral regions. All calves were sacrificed one hour later to confirm the location of injected drug. The drug was found in a muscle layer when injected with a needle 15 mm long to the following positions, 1. the midpoint between the central position of the gluteal region (CG) and the tuber coxae (M-CTc), 2. the midpoint between CG and the tuber ossis ischii (M-CTo), 3. the central position of M. semimembranaceus in the femoral region (CF). Seven calves, weighing 130-150 kg, were given intramuscularly 5 ml of ABPC suspensions at M-CTo and CF and sacrificed one hour (4 calves) and 3 days (3 calves) later. ABPC diffused along the long axis of the muscle fibers but not to the radial direction. ABPC was detected only in the injected muscle layer even after 3 days indicating that the drug did not diffuse to the neighboring muscles. In the injected muscle layer, concentration of ABPC was remarkably different from part to part. From these results, sampling of the injected muscle for the drug residue study was proposed as follows: 1. isolate about 100 g of muscle just under the stick point marked on the skin considering the direction of drug diffusion, and 2. isolate separately about 200 g of the surrounding muscle to confirm if the sampling is appropriate.