Cloning of a specific DNA region of white top pathogen of pea and its detection by polymerase chain reaction     

Ayumi?Suzuki  Yuichi?TakikawaEmail author 《Journal of General Plant Pathology》2004,70(3):174-180

White top strain (WT strain) of Pseudomonas syringae pv. pisi (Ppi) is a variant strain causing white top disease of peas. The WT strain is distinguishable from common Ppi strains only by symptom expression chlorosis and whitening of apical shoots. To develop a specific detection method for the WT strain, we cloned a specific DNA region of the WT strain using transposon tagging. Five mutants defective in white top symptom expression were obtained. A part of the Tn5-flanking region was cloned and labeled as a hybridization probe. One clone, pAY3, gave two signal bands, one of which was detected from the genomic DNA of all the WT and the common Ppi strains; another was specific to WT strains. A restriction map of pAY3 showed that it contains two BamHI fragments; one is 5.0kb in length involving a part of Tn5, and the other is 1.5kb, did not carry Tn5, and may have been accidentally ligated into pAY3. The 1.5-kb band was subcloned as pAY13 and was used as a probe. It hybridized specifically to WT strains. These results suggested that the WT strains have a specific DNA region and that part of the region was successfully cloned. Sequence analysis of pAY13 showed that it is similar to part of nonribosomal peptide synthetases (NRPSs) genes. The deduced amino acid sequence of pAY13 suggested the existence of eight conserved motifs of NRPSs. WT strain-specific PCR primers, PS1 and PS2, were designed from the DNA sequence. These primers gave a specific amplification product of 981bp from both the genomic DNA and a direct cell preparation of WT strains. No specific amplicon was produced from Ppi strains that caused only water-soaked lesions or from strains of other P. syringae pathovars. A specific amplicon was not produced from four strains of the pea pathogen: P. marginalis pv. marginalis, P. viridiflava, Erwinia carotovora ssp. carotovora, Xanthomonas campestris pv. pisi. Using the primers, WT strain was detected from water-soaked lesions and green and white tissues without water soaking.The sequence reported in this paper has been deposited in the DDBJ database under accession no. AB117755  相似文献   

Characteristics of a hepatozoonosis in lungs of Japanese black bears (Ursus thibetanus japonicus)     

Uni S  Matsubayashi M  Ikeda E  Suzuki Y 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2003,65(3):385-388

In a survey of pathologic agents in the wild animals of central Japan, we found a Hepatozoon sp. in the lungs of Japanese black bears in Fukui, Shiga, and Gifu Prefectures, Japan. Histopathologic examination of organs and tissues from the 18 bears inspected showed hepatozoonosis in all. Immature and mature meronts were found in all lobules and between alveoli, with a few found between pleura and in connective tissue. In the lungs, inflammatory cells were not found around meronts, merozoites, or tubercles made of macro-phages including zoites, but inflammatory cells were found around degenerating cells, zoites, and tubercles. A Hepatozoon sp. has not been reported as being detected in bears of any species before.  相似文献   

Age-related change and allometry of skull and canine of sea otters,Enhydra lutris     

Hattori K  Burdin AM  Suzuki M  Ohtaishi N 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2003,65(4):439-447

Skulls and canines of 460 sea otters from Lopatka Cape, Kamchatka, were examined to assess development patterns, individual variation and sexual differences. An allometric formula was applied to morphometrical data, and the relative growth of each character to total length of skull was analyzed. In both sexes, most morphometrical characters ceased growth at about 2 years of age. Canine root length increased rapidly during the first year of life, while crown length decreased due to remarkable wear. There was large individual variation in the feeding and breathing/sniffing apparatus, while there was little variation in braincase size. There were sexual differences in most characteristics, although males and females showed similar growth patterns. The coronoid process of the mandible showed positive allometry in both sexes, and we attributed this finding to feeding habits. The fact that only male mastoids showed positive allometry may be due to the need for male otters to maintain a passing territory.  相似文献   

Inhibin secretion in the golden hamster (Mesocricetus auratus) testis during active and inactive states of spermatogenesis induced by the restriction of photoperiod     

Kawazu S  Kishi H  Saita E  Jin W  Suzuki AK  Watanabe G  Taya K 《The Journal of reproduction and development》2003,49(1):87-97

Gonadal function in the male golden hamster (Mesocricetus auratus) was investigated during exposure to a short photoperiod condition. Within 3 weeks of exposure to the short photoperiod condition, FSH and testosterone in the plasma significantly decreased, and subsequently immunoreactive (ir)-inhibin significantly decreased. Testicular contents of ir-inhibin and testosterone, and pituitary contents of LH and FSH also significantly decreased by 3 weeks with regression of weight of testes, epididymis and seminal vesicles and sperm head count. Circulating LH varied but not significantly. Thereafter, all reproductive parameters and secretion of LH, FSH, ir-inhibin and testosterone gradually recovered after 17 weeks of exposure even though animals continued to be subjected to the short photoperiod condition. Plasma concentrations of inhibin B and inhibin pro-alphaC were detectable and were significantly decreased after 15 weeks of exposure to the short photoperiod, but their levels were still detectable. Immunopositive reaction of inhibin alpha and betaB subunits was found in Sertoli cells and Leydig cells in the regressed testes of animals subjected to short photoperiod as was also seen in animals before exposure to the short photoperiod. Although the spermatogenic cycle was suppressed like prepubertal animals, the present study showed that the testicular recovery, so-called refractoriness, is functionally different from the developing stage of immature animals, especially with regard to inhibin secretion. The present results showed that changes in FSH preceded changes in inhibin during the regression and recovery phases, indicating that FSH is a major regulatory factor of inhibin secretion in male golden hamsters. The present study also demonstrated that regressed testes still secrete a small amount of bioactive inhibin during exposure to a short-photoperiod condition.  相似文献   

Expression analyses of sex steroid-regulated genes in neonatal rat hypothalamus     

Yonehara K  Suzuki M  Yamanouchi K  Nishihara M 《The Journal of reproduction and development》2003,49(6):547-552

Estrogen plays an important role in sexual differentiation of the brain in rats during the perinatal period. To elucidate molecular mechanisms underlying sexual differentiation of the brain, in this study we investigated genes differentially expressed between sexes or induced to express by estrogen in neonatal rat hypothalamus using DNA microarray analysis in combination with real-time RT-PCR. It was found that the levels of expression of the genes encoding glutamic acid decarboxylase 65 and coronin 1b were higher in male than female hypothalamus on postnatal day (PN) 5 and those of collagen type 3 alpha1 and thioredoxin reductase 2 genes in female hypothalamus on PN5 were decreased and increased, respectively, by treatment with estradiol on PN2. Then the developmental changes in the expression of these 4 genes were examined from 1 day before the parturition to PN9, and they all showed sexual dimorphic patterns. In addition, dependence of the expression of these genes on either estradiol, testosterone or dihydrotestosterone during the neonatal period was confirmed. These results suggest that these four genes are involved in sexual differentiation of the rat brain, and that androgen per se as well as estrogen may take part in the processes.  相似文献   

Generation of transgenic rats expressing enhanced green fluorescent protein in gonadotropin-releasing hormone neurons   总被引：3，自引：0，他引：3  

Fujioka H  Suzuki M  Yamanouchi K  Ohta A  Nagashima H  Kato M  Nishihara M 《The Journal of reproduction and development》2003,49(6):523-529

Hypothalamic gonadotropin-releasing hormone (GnRH) neurons govern reproductive function by controlling the release of gonadotropins from the pituitary. To facilitate identification of living GnRH neurons, here we attempted to generate transgenic rats that express enhanced green fluorescent protein (EGFP) in GnRH neurons. About 3 kb of rat GnRH promoter region was inserted into the EGFP reporter cassette, and the expression of EGFP fluorescence was confirmed in several cell lines following transient transfection. Then we successfully generated a transgenic rat by injecting linearized GnRH-EGFP transgene into the pronuclei of fertilized oocytes. The GnRH-EGFP transgenic rats expressed EGFP in the brain, but not in the ovary, testis or thymus. Immunohistochemical examination revealed that detectable EGFP fluorescence was confined to the cell body of GnRH-immunoreactive neurons in the septum and preoptic area, while no EGFP signal was discernible in the median eminence where abundant GnRH-immunoreactive fibers were observed. The mean percentage of EGFP-positive cells in the GnRH-positive cells was 76.3%. The GnRH-EGFP transgenic rats generated in the present study will enable characterization of properties of individual GnRH neurons.  相似文献   

Renal lesions in spontaneous insulin-dependent diabetes mellitus in the nonobese diabetic mouse: acute phase of diabetes     

Maeda M  Yabuki A  Suzuki S  Matsumoto M  Taniguchi K  Nishinakagawa H 《Veterinary pathology》2003,40(2):187-195

The nonobese diabetic mouse is a model of spontaneous insulin-dependent diabetes mellitus. The present study made longitudinal observations of renal lesions in the acute-progressive phase of diabetic mice 0, 10, 20, 30, and 40 days after onset of diabetes without insulin therapy. Plasma creatinine and blood urea nitrogen concentrations gradually increased after onset of diabetes. Kidney weight increased and plateaued at day 20. Under electron microscopy the glomeruli demonstrated only mild changes on day 40. In the proximal tubules proliferating cell nuclear antigen-positive nuclei and nuclear divisions were increased on days 10 and 20. On day 40 of diabetes, increased periodic acid-Schiff-positive granules, confirmed as lysosomal dense bodies, increased neuronal nitric oxide synthase (nNOS) positive reaction, and decreased periodic acid-Schiff staining in the brush border were observed in the proximal straight tubules. In the juxtaglomerular apparatus stratified macula densa were decreased with time in diabetes compared with the findings on day 0, and this macula densa positively reacted with nNOS. No changes in renin levels were observed. In addition, apoptotic cells were not detected. In conclusion, this research represents the first thorough characterization of acute changes in nonobese diabetic mouse kidneys. The results demonstrated renal hypertrophy and slight glomerular injury in early stages and structural alteration of the proximal straight tubules at later stages during the acute phase of diabetes. Furthermore, increased nNOS may represent one of the pathogenic factors of diabetic nephropathy.  相似文献   

EXTRA-ABDOMINAL CHONDROLIPOMA IN A DOG     

Shigeyuki Tanabe  Kazutaka Yamada  Yoshiyasu Kobayashi  Koudai Suzuki  Miho Yamaguchi  Yuji Uzuka  Takao Sarashina  Michiyo Inoue 《Veterinary radiology & ultrasound》2005,46(4):306-308

A 10-year-old male maltise dog was presented for evaluation of a gradual abdominal enlargement, which had developed over 2 years. (On the radiograph of the abdomen, a significant distention of the abdomen with a large accumulation of fat could be seen.) The mass effect was noticeable because of the rightward displacement of the viscera. There was multifocal amorphous mineralization, mostly in the right abdomen. Computed tomography (CT) identified a 13 x 13 x 10 cm mass, most likely an intra-abdominal lipoma, by which the digestive tract had been displaced in the abdomen. It was only during surgery, however, that the mass was found to be located between the peritoneum and transversus abdominis muscles and the rectus abdominus muscle. It was completely resected. Histopathological examination revealed that the mass was chondrolipoma. The amorphous radiopaque lesion on the images corresponded to cartilaginous tissues with calcification. There has been no recurrence for 10 months. This is the first report of an extra-abdominal chondrolipoma in dogs. Cartilaginous metaplasia of lipoma, located in the abdominal area, should be included in the differential diagnosis for multifocal amorphous mineralization.  相似文献   

Antibabesial activity of protoberberine alkaloids and 20-hydroxyecdysone from Arcangelisia flava against Babesia gibsoni in culture     

Subeki  Matsuura H  Takahashi K  Yamasaki M  Yamato O  Maede Y  Katakura K  Suzuki M  Trimurningsih  Chairul  Yoshihara T 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2005,67(2):223-227

Bioassay-guided fractionation of the boiled extract from the stems of Arcangelisia flava led to the isolation of palmatine (1), berberine (2), jatrorrhizine (3), dihydroberberine (4) and 20-hydroxyecdysone (5). The chemical structures of these compounds were elucidated on the basis of their chemical and spectral evidence. The isolated compounds were evaluated for their growth inhibiting effects on Babesia gibsoni in culture for a week. Compounds (1-4) showed significant inhibitions at concentrations from 100 to 1.0 microg/ml, while compound 5 at a concentration of 100 microg/ml, only.  相似文献   

A comparative histological study on the distribution of striated and smooth muscles and glands in the esophagus of wild birds and mammals     

Shiina T  Shimizu Y  Izumi N  Suzuki Y  Asano M  Atoji Y  Nikami H  Takewaki T 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2005,67(1):115-117

Musculature and glands of the esophagus in various wild birds and mammals were examined histologically. Cervical and thoracic esophagi of all birds used (mallard, spot-billed duck, Ural owl and Hodgson's hawk-eagle) were comprised of smooth muscle fibers only. In contrast, esophagi of the nutria, Japanese raccoon dog, common raccoon and Japanese marten consisted largely of striated muscle fibers. In the masked palm civet, Japanese macaque and bottlenose dolphin, esophageal muscle layers consisted of both striated and smooth muscle fibers. Esophageal glands were observed except for the nutria and masked palm civet. These results show a wide variety of the structural composition in the esophagus of wild animals, particularly mammals, examined in this study.  相似文献