Discrimination of three amphistome species by PCR-RFLP based on rDNA ITS2 markers

Three bovine amphistomes, Calicophoron calicophorum, Orthocoelium streptocoelium and Homalogaster paloniae, are common in Japan. This study was carried out to describe ITS2 sequences in the 3 species, and to evaluate a PCR-RFLP technique based on ITS2 sequence for the species identification in single eggs of the parasites. The ITS2 sequences of the three species contained 19 variable sites including one gap. The sequence difference between species was 4.2-5.3%. The three species of amphistomes were identified based on the difference in the restriction sites of Acc II on the ITS2 sequence.     

Survey on wild rodents for endoparasites in Iwate Prefecture, Japan

Wild rodents (58 Apodemus speciosus, 29 A. argenteus and 7 Microtus montebelli) were surveyed for endoparasites in Iwate Prefecture, Japan, from October to December 1995 and from April to October 1996. Two trematodes (Echinostoma macrorchis, Plagiorchis muris), 4 or more cestodes (Hymenolepis diminuta, Raillietina coreensis, Cladothyridium spp., Cysticercus fasciolaris), 12 nematodes (Carolinensis minutus, Eucoleus sp., Heligmosomoides kurilensis, H. protobullosus, H. speciosus, Heterakis spumosa, Rhabditis (Pelodera) orbitalis, Rictularia cristata, Syphacia emileromani, S. frederici, S. montana, Trichuris sp.) and 3 protozoans (Giardia sp., Trichomonas sp., Trypanosoma sp.) were identified. The two species of Apodemus were similar to each other, but they were extremely different from M. montebelli in parasite fauna.     

剪股颖属植物遗传分化及系统关系的分子标记研究

剪股颖属植物形态变异大、倍性复杂、种间易于杂交,导致异名多、分类混乱。本研究采用4种分子标记技术,对包括匍茎剪股颖、巨序剪股颖及红顶草在内的7种剪股颖,共58个个体的遗传分化和系统关系进行研究。根据谱带清晰程度及多态性,筛选出9个RAPD引物、2个SSR引物、5个ISSR引物及1个SCAR标记,并建立了最适的PCR反应条件。采用最大简约法和距离法对7种剪股颖植物进行系统树分析,并用靴带检验法计算内部分支的支持率。启发式搜索得到的简约树和由UPGMA方法得到的表征树近乎相同,其中,匍茎剪股颖和红顶草构成单系类群并得到100%的置信支持。红顶草显示了匍茎剪股颖特异性特征谱带,但同时具有不同于匍茎剪股颖的遗传分化,支持将红顶草视为匍茎剪股颖一个变种的观点。对匍茎剪股颖与巨序剪股颖间的遗传分化进行了探讨。     

Estimation of the carcass composition from a cross-section of the rib-loin from crossbred Japanese Black × Limousin F2 cattle by computer image analysis

The carcass composition of crossbred Japanese Black × Limousin F2 cattle was examined in order to find an accurate carcass composition equation. The test animals included 17 steers and 17 heifers. The 28 image measurements from the area encircling the vertical line to the thoracic vertebra and the line from the thoracic vertebra between the sixth and seventh rib‐bones were measured by computer image analysis. The relationships between the 29 parameters that added the carcass left side weight of the animal and the carcass composition were suggested. The carcass composition included muscle weight, muscle ratio, fat weight and fat ratio. The carcass composition from steers was estimated by an equation composed of these three or four parameters (R² = 90.80%, 79.30%, 90.75% and 73.70%, respectively). The selected parameters were measured without cutting the thoracic vertebra. The carcass composition from heifers was estimated by an equation composed of two to four parameters (R² = 96.15%, 90.98%, 93.60% and 88.22%, respectively). The parameters for the estimation of the muscle and fat weight, and muscle and fat ratio are very similar. Furthermore, the equations using the parameters could estimate the carcass composition from the Japanese Black × Limousin cattle resource population.     

Suppressive effect of liquid potassium silicate on powdery mildew of strawberry in soil

A soil drench of soluble silicon (liquid potassium silicate) was tested for the control of powdery mildew of strawberry in soil for 2 years in a vinyl house under growers' conditions on two cultivars: very susceptible Toyonoka and slightly susceptible Sachinoka. Soluble silicon suppressed the disease more effectively as a preventive control than as a control to reduce initial incidence. The suppressive effect against this disease was more effective for Toyonoka than for Sachinoka. The control value of soluble potassium silicate was 85.6% for Toyonoka in the first year and 60.2% in the second year, while for Sachinoka it was 58% in the first year and 40.6% in the second year. Strawberry leaf hardness was measured for the control and silicate-treated leaves. Although silicate-treated leaves seemed to be harder than control leaves, leaf hardness did not vary significantly between controls and silicate-treated leaves in either of the 2 years, and the mode of preventive action of liquid potassium silicate remains unknown. A factor other than a physical action may be involved.     

Pea extracellular Cu/Zn-superoxide dismutase responsive to signal molecules from a fungal pathogen

We previously reported that the release of O₂ ⁻ from isolated pea cell walls was enhanced by a 70-kDa glycoprotein elicitor but was suppressed by mucin-type glycopeptide suppressors (supprescins A and B) prepared from pycnospore germination fluid of Mycosphaerella pinodes, causal agent of Mycosphaerella blight of pea. Here, we show that superoxide dismutase (SOD) in the apoplast fluid/cell wall of pea seedlings responds to the fungal elicitor and suppressor molecules. In a pharmacological study and with internal amino acid sequencing, the apoplastic SOD in a pea cultivar Midoriusui was found to be a Cu/Zn type SOD. We cloned a full-length cDNA of the Cu/Zn-SOD and designated it as PsCu/Zn-SOD1. An increase in PsCu/Zn-SOD1 mRNA and the PsCu/Zn-SOD1 protein was induced by treatment with the elicitor more intensively than by wounding. Such induction by the elicitor or wounding, however, was inhibited by the concomitant presence of supprescins. The SOD activity of recombinant PsCu/Zn-SOD1 was regulated directly by these signal molecules in a manner similar to their effect on the SOD activity in the apoplastic fluid and in the cell wall-bound proteins. Based on these findings, we discuss a role for PsCu/Zn-SOD1 in the pea defense response. The nucleotide sequence data of PsCu/Zn-SOD1 reported are available in the DDBJ/EMBL/GenBank databases under accession number AB189165.     

Caprine somatic cell nuclear transfer using in vivo matured oocytes collected by laparoscopic follicular aspiration

In vivo matured oocytes collected by laparoscopic follicular aspiration (LFA) from hormone treated female goats were used as recipient ooplasts for somatic cell nuclear transfer (SCNT). Japanese native (Shiba) goats were used as donor females and some donor females were used repeatedly (two or three times) at intervals of a few months. To induce synchronization of estrus, a sponge containing 0.5 g of progesterone was inserted into the vagina of each goat for 14 days. These animals were also treated with follicle stimulating hormone (FSH) in a series of 8 injections over 4 days. The first FSH injection was administered on the morning of day 9 of sponge insertion. On the morning of day 13, 50 µg of gonadotropin‐releasing hormone (GnRH) was injected into each animal. Twenty‐nine hours after GnRH injection, LFA was performed. After removal of cumulus cells, collected oocytes with the first polar body were selected and enucleated for nuclear transfer. Anterior pituitary cells isolated from an adult male Shiba goat were transfected with a DNA fragment containing the enhanced green flourescent protein gene and the puromycin resistance gene. A single donor cell was inserted into the perivitelline space of each enucleated oocyte and fusion was induced with one electric pulse of 20 V for 10 µs. The SCNT goat eggs were cultured in chemically defined medium at 38.5°C in 5% CO₂, 5% O₂, 90% N₂ for 9 days. By LFA, 396 oocytes were collected from a total of 30 females. After removal of cumulus cells, 64% of them extruded the first polar body. The percentage of SCNT goat eggs produced using in vivo matured oocytes which developed to the blastocyst stage (20–21%) was significantly higher (P < 0.05) than that produced with in vitro matured oocytes (3–8%).