Identification of genes expressed during spore germination of Mycosphaerella pinodes

Mycosphaerella blight, caused by Mycosphaerella pinodes, is one of the major diseases of cultivated pea (Pisum sativum L.). To isolate the genes that are up- and down-regulated during spore germination, suppression subtraction hybridization (SSH) was performed between ungerminated and germinated spores. The 232 and 128 clones from forward and reverse libraries, respectively, were collected, sequenced, and analyzed with a BLASTX homology search. About 95% of the 32 selected clones were expressed during spore germination on a paper sheet and during infection of pea leaves. We discuss the applicability of the SSH libraries for analyzing M. pinodes genes involved in the early stage of infection.     

Molecular Characterization of <Emphasis Type="Italic">Bean yellow mosaic virus </Emphasis>from Vegetatively Propagated Dwarf <Emphasis Type="Italic">Gentiana </Emphasis>Plants

The causative virus (isolate No. 4) of gentian (Gentiana spp.) mosaic, which had been identified previously as Clover yellow vein virus (C1YVV) on the basis of host range and serological reactions, was re-identified as Bean yellow mosaic virus (BYMV) on the basis of the nucleotide sequences of the gene for the coat protein (CP) and the 3′-noncoding region, as well as the predicted amino acid sequence of CP. Received 16 April 2002/ Accepted in revised form 19 June 2002     

Establishment of a canine lens epithelial cell line

Some immortalized lens epithelial cell lines have been established and are useful for molecular analysis. The establishment of additional cell lines must, however, enable a variety of in-vitro examinations. The objective of this study was to establish a new canine lens epithelial cell line by isolating CLC-1 cells from the lens tissue of a dog with cataracts. In CLC-1 cells, transforming growth factor beta (TGF-β) treatment significantly decreased gene expression of an epithelial marker and elevated that of mesenchymal markers; these characteristics are similar to those of a human lens epithelial cell line. Interestingly, CLC-1 cells exhibited lower expression of an epithelial marker and higher expression of mesenchymal markers than an anterior lens capsule. These results suggest that CLC-1 cells were derived from a cell population that was committed to epithelial-mesenchymal transition in cataract lens tissue. In conclusion, CLC-1 cells could be useful for analyzing molecular pathogenesis in canine cataracts.     

Pedigree analysis of genetic subdivision in a population of Japanese Black cattle

Twenty‐five subpopulations (i.e. populations of prefectures) of more than 2000 Japanese Black cows younger than or equal to 10 years of age were analyzed to evaluate the genetic relationships in the current population. The total number of cows analyzed was 392 346 and their pedigrees were traced back to 1944 or before. Using the pedigree records, the genetic relationships among the subpopulations were estimated by the two different measurements: (i) the average additive relationship coefficients and (ii) Nei's standard genetic distances. Principal component analysis (PCA) was performed to the matrix of the average additive relationship coefficients, and the factor loadings of subpopulations were plotted on the plane to visualize the genetic configuration of subpopulations. To understand the grouping process of the subpopulations, cluster analysis was applied to the matrix of the Nei's genetic distances, and a dendrogram was constructed. There was a high consistency between the results from PCA and cluster analysis. Eight subpopulations with relatively low migration rates showed their unique genetic compositions, and the other 17 subpopulations with high migration rates formed a single cluster. The major cause of the genetic similarity among the 17 subpopulations was inferred to be the strong genetic influence from one subpopulation (Hyogo prefecture) with prominent characteristics for meat quality.     

Effects of supplemental ascorbic acid on performance, organ weight and plasma cholesterol concentration in broilers treated with propylthiouracil

1. Four experiments were conducted to determine if dietary ascorbic acid (AA) affects body weight gain, food intake, organ weights, plasma cholesterol concentration, and ascorbic acid concentration in the plasma and liver of growing male broilers treated with an antithyroidal agent, propylthiouracil (PTU). 2. In the first experiment, 15 mg AA was administered daily into the crop of chicks fed on a diet supplemented with or without PTU (500 mg/kg). Administration of AA reduced plasma cholesterol concentrations in the PTU-treated chicks. 3. In the other three experiments, chicks were given the basal diet or an AA-containing (3 g/kg) diet supplemented with or without PTU (250 mg or 500 mg/kg). Feeding AA partly prevented the decreases in body weight gain, gain:food ratio and weights of the bursa of Fabricius and thymus in chicks fed on the 250 mg/kg PTU diet, and also prevented the increase in plasma cholesterol concentrations in chicks fed on the PTU diet. 4. These results suggest that AA improves the performance of chicks with experimentally induced hypothyroidism.     

Complete cDNA sequence and mRNA expression of dog preproendothelin-3

The full-length cDNA of dog preproendothelin-3 (PPET3) was cloned from lung tissue using RT-PCR and rapid amplification of cDNA ends. Aside from the poly (A) tail, the full-length cDNA was 1976 bp. A polyadenylation signal sequence and one copy of a consensus sequence, ATTTA, which is related to mRNA turnover, was found in the 3' noncoding region. The cDNA had a 594-bp open reading frame encoding a 198-amino acid polypeptide. Regions corresponding to a bioactive mature ET3 peptide, an intermediate form known as big-ET3, and an ET3-like peptide were observed in dog PPET3. Expression of PPET3 mRNA was detected throughout the organs examined, which included heart, lung, liver, kidney, spleen, stomach, pancreas, duodenum, colon, uterus, ovary and testis.     

Susceptibility to N-methyl-N-nitrosourea-induced retinal degeneration in different rat strains

To evaluate the potential role of genetic background in the susceptibility to retinal degeneration induced by N-methyl-N-nitrosourea (MNU), female rats of the Sprague-Dawley (SD), Long-Evans (LE) and Copenhagen (CH) strains were administered 50 mg/kg MNU or saline at 7 weeks of age. Retina morphology and morphometric analysis of all rats was performed 7 days after MNU administration. Atrophy of both the peripheral and central outer retina occurred in all rat strains exposed to MNU. Decreased photoreceptor cell ratio and increased retinal damage ratio were observed. The severities of the retinal atrophy were similar among all three rat strains. In conclusion, MNU-induced photoreceptor degeneration developed consistently in all three strains regardless of the absence (SD rats) or presence (LE and CH rats) of melanin in the retina, suggesting that genetic and melanin factors did not affect photoreceptor cell death after MNU.     

Possible altitude and temperature limits on pine wilt disease: the reproduction of vector sawyer beetles (Monochamus alternatus), survival of causal nematode (Bursaphelenchus xylophilus), and occurrence of damage caused by the disease

Pine wilt disease is caused by the pine wood nematode [Bursaphelenchus xylophilus (Steiner et Buhrer) Nickle]. In East Asia, an important vector of the nematode is Monochamus alternatus Hope. We determined the tolerance and reproductive ability of sawyer beetles and the nematode to altitude and temperature at elevations between 850 and 1,450 m on Mt. Fuji in Japan. The number of emergent adults decreased markedly along the altitudinal gradient, but the beetle could still reproduce at 1,050 m (8.2 °C annual mean temperature). Beetles with a 2-year life cycle increased rapidly in number with increasing altitude. The pine wood nematode survived through winter at all altitudes tested (850–1,450 m). The beetle population decreased between 950 (9.1 °C) and 1,150 m (8.3 °C). Therefore, the beetle population seems to be stable at 850 m (10.2 °C) and lower altitudes (higher temperatures) but cannot be maintained from 950 (9.1 °C) to 1,150 m (8.3 °C) without constant immigration of beetles from lower altitudes. The beetles could not reproduce at altitudes above 1,150 m (lower than 8.2 °C). From the mean and effective cumulative temperatures, we concluded that the beetle (and its population) can endure temperatures lower than those previously reported. Pine wilt disease also occurred at lower temperatures and higher altitudes than expected. We have summarized the principal strategies for controlling the disease at high altitudes based on these results.