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71.
Human ribosomal protein (RP) gene sequences with respect to intron/exon structures and corresponding cDNA or genomic data of fish species were obtained from the GenBank database. Based on conserved exon sequences, 128 primer pairs for 41 genes were designed for exon-primed intron-crossing (EPIC) polymerase chain reaction (PCR). In reference to the draft genome sequences of the Pacific bluefin tuna (Thunnus orientalis), 12 primer pairs expected to amplify introns of the bluefin tuna with lengths of 500–1000 bp were selected and applied to six distantly related fish species belonging to the Orders Clupeiformes, Tetraodontiformes, Pleuronectiformes, Perciformes, Scorpaeniformes, and Anguilliformes. PCR amplification was observed for at least four species in each primer pair, and all fragments were larger than those expected for intronless amplification. Single fragment amplification was observed for at least seven primer pairs per species. Fragment sizes of the bluefin tuna for nine primer pairs corresponded to those expected from the genomic data. Thus, our primer pairs are potentially applicable to a wide variety of fish species and serve as an initial step for isolating single-copy nuclear DNA sequences.  相似文献   
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73.
Ethylene biosynthesis in kiwifruit, Actinidia chinensis ‘Sanuki Gold’ was characterized using propylene, an ethylene analog, and 1-methylcyclopropene (1-MCP), an inhibitor of ethylene perception. In fruit harvested between a young stage (66 days after pollination) (DAP) and an early commercial harvesting stage (143 DAP), 2 days of exposure to propylene were sufficient to initiate ethylene biosynthesis while in fruit harvested at commercial harvesting stage (154 DAP), 4 days of propylene treatment were required. This observation suggests that response of ethylene biosynthesis to propylene treatment in kiwifruit declined with fruit maturity. Propylene treatment resulted in up-regulated expression of AC-ACO1, AC-ACO2, AC-SAM1 and AC-SAM2, prior to the induction of AC-ACS1 and ethylene production, confirming that AC-ACS1 is the rate limiting step in ethylene biosynthesis in kiwifruit. Treatment of fruit with more than 5 μL L?1 of 1-MCP after the induction of ethylene production subsequently suppressed ethylene production and expression of ethylene biosynthesis genes. Treatment of fruit with 1-MCP at harvest followed with propylene treatment delayed the induction of ethylene production and AC-ACS1 expression for 5 days. These observations suggest that in ripening kiwifruit, ethylene biosynthesis is regulated by positive feedback mechanism and that 1-MCP treatment at harvest effectively delays ethylene production by 5 days.  相似文献   
74.
The relative parasitic fitness of isolates of Pyricularia oryzae with different sensitivities to isoprothiolane (di-isopropyl 1,3-dithiolan-2-ylidenemalonate) and IBP (S-benzyl O, O-di-isopropyl phosphorothioate) was studied in the absence of fungicides. Four field isolates (S-1,S-2,MR-1 and MR-2) and two in-vitro mutants (Rvt-1 and Rvt-2) were used for disease epidemics. S-l and S-2 were wild types; MR-1 and MR-2 were sensitive to isoprothiolane and moderately resistant to IBP, and Rvt-1 and Rvt-2 were resistant to both fungicides. Twelve epidemics were made by inoculating rice seedlings with mixed conidial suspensions of two isolates or mutants. The value of relative parasitic fitness (W = 0-1.0) was calculated for each isolate and epidemic. S-l and S-2 were stronger (W = 1.0) than MR-2, Rvt-1 and Rvt-2; but weaker (W = 0.75, 0.73, respectively) than MR-1. MR-1 was strongest among all isolates and mutants used. MR-2 was slightly weaker (W = 0.9-1.0) than S-l and S-2, but stronger than Rvt-1 and Rvt-2. Rvt-1 and Rvt-2 had smaller values of W, ranging from 0.25-0.58, in the epidemics with each field isolate. These results suggest that the proportions of in-vitro mutants do not increase unless intensive selection pressure is given, and would be expected to decrease rapidly after the selection pressure is removed. Isolates moderately resistant to IBP, such as MR-1 and MR-2, however, had high values of W, suggesting that they would increase or would not decrease rapidly in the absence of selection pressure. These results may well explain why isolates highly resistant to isoprothiolane and IBP have seldom been found, and why a large number of isolates moderately resistant to IBP but sensitive to isoprothiolane have been observed in the field.  相似文献   
75.
Restriction enzyme-mediated integration (REMI) mutagenesis was used to isolate mutants of Fusarium oxysporum f. sp. melonis impaired in pathogenicity. The race 2 strain Mel02010 was transformed with linearized pSH75, conferring resistance to hygromycin B, with or without the enzyme used to linearize the plasmid. Addition of restriction enzymes did not affect the transformation frequency. A total of 2929 REMI transformants were tested for pathogenicity to three melon cultivars, Amus, Ogon 9 and Ohi. The race 2 strains are pathogenic to Amus and Ogon 9, but not to Ohi. Of 43 transformants with reduced pathogenicity on susceptible melon cultivars, 12 mutants were examined in detail for pathogenicity, vegetative growth and integrative mode of pSH75. The levels of pathogenicity varied among these mutants. Two mutants (B48 and B137) almost completely lost pathogenicity to both susceptible cultivars, and the others had reduced pathogenicity. Mutants B48, B241, B886 and X36 were also impaired in vegetative growth. Mutant B809 was a biotin auxotroph. By DNA gel blot analysis, nine mutants were found to contain a single copy of the transformation vector. These mutants may thus be useful in isolating genes involved in pathogenicity. Received 22 December 2000/ Accepted in revised form 16 April 2001  相似文献   
76.
The effects of a novel pyrimidine herbicide, NS-245852 [2-chloro-6-fluorophenyl-4-(trifluoromethyl)thieno[2,3-d]pyrimidine-2-yl-ketone], on mitosis in oat ( Avena sativa L. cv. Zenshin) root tips were investigated by using light and immunofluorescence microscopy. The root growth was strongly inhibited at 10−7 mol L−1 of NS-245852, and swollen root tips were induced at 5 × 10−8 mol L−1. As observed by the use of light microscopy, the herbicide produced disrupted mitosis and large polynucleate cells in the meristematic root tissue. These symptoms were similar to those of mitotic disrupter herbicides. The immunofluorescence microscopy studies of the root tip cells treated for 30 min revealed that spindle fibers and the preprophase band were reduced, although kinetochore fibers and the phragmoplast were not affected. Kinetochore fibers remained as small fluorescence spots, and the phragmoplast disappeared after a 3 h treatment. No microtubule arrays were observed by a longer treatment (longer than 3 h). Among the microtubule arrays, spindle fibers and the preprophase band were found to be the most sensitive to the herbicide, whereas kinetochore fibers were the most resistant. The phragmoplast was intermediate. Thus, the primary action of NS-245852 is the inhibition of polymerization of tubulin into microtubules.  相似文献   
77.
The alpha antigen, which is an immunodominant antigen, is a 30 kDa protein secreted by mycobacterial species. The C-terminal regions of alpha antigens are quite divergent. We investigated the question of whether the C-terminal regions of Mycobacterium avium alpha antigen (A-alpha), M. intracellulare alpha antigen (I-alpha) and M. bovis BCG alpha antigen (B-alpha) contained species-specific B-cell epitopes. We investigated the reactions of these peptides with anti-A-alpha, anti-I-alpha and anti-B-alpha sera prepared from BALB/c in a Western blot assay and ELISA. The C-terminal regions of I-alpha reacted exclusively with anti-I-alpha serum. The results of the inhibition assay of antibodies binding to I-alpha by peptides of C-A-alpha, C-I-alpha, and C-B-alpha are that only C-I-alpha inhibited the binding of antibodies to C-I-alpha. We found that the C-terminal region was B-cell epitope-specific to I-alpha in BALB/c mice.  相似文献   
78.
A pinealoma (benign) was found in a 61-week-old male Crj:CD (SD) IGS rat. The neoplasm was located between the cerebral hemispheres and the cerebellum. Histologically, the tumor cells consisted of two cell types: large, pale-staining cells and small dark-staining cells. A fibrovascular stroma divided the tumor cells into incomplete lobules or nest structures. Relatively numerous mitoses were noted in the tumor cells. Ultrastructurally, the tumor cells contained dense-cored vesicles, approximately 120 nm in diameter.  相似文献   
79.
A 5-month-old, male, Shih Tzu dog manifesting polyuria and polydipsia since 2-month-old was presented to our hospital with additional clinical complaints of vomiting and depression during recent a few days. Despite the symptomatic therapy for chronic renal failure, he died on the day after medication. Macroscopically, both kidneys were small in size with rough surface. Microscopical examination revealed bilateral renal fibrosis with dysplastic changes consisting of immature glomeruli and tubules, and foci of adenomatoid proliferation of tubular epithelium. In addition, incomplete lobulation of medulla with pelvic structures was also noticed in the right kidney. From these findings, the present case was diagnosed as renal dysplasia in Shih Tzu dog which was documented in the literatures.  相似文献   
80.
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