RNA silencing as a tool to uncover gene function and engineer novel traits in soybean     

Megumi Kasai  Akira Kanazawa 《Breeding Science》2012,61(5):468-479

RNA silencing refers collectively to diverse RNA-mediated pathways of nucleotide-sequence-specific inhibition of gene expression. It has been used to analyze gene function and engineer novel traits in various organisms. Here, we review the application of RNA silencing in soybean. To produce soybean lines, in which a particular gene is stably silenced, researchers have frequently used a transgene that transcribes inverted repeats of a target gene segment. Suppression of gene expression in developing soybean embryos has been one of the main focuses of metabolic engineering using transgene-induced silencing. Plants that have enhanced resistance against diseases caused by viruses or cyst nematode have also been produced. Meanwhile, Agrobacterium rhizogenes-mediated transformation has been used to induce RNA silencing in roots, which enabled analysis of the roles of gene products in nodulation or disease resistance. RNA silencing has also been induced using viral vectors, which is particularly useful for gene function analysis. So far, three viral vectors for virus-induced gene silencing have been developed for soybean. One of the features of the soybean genome is the presence of a large number of duplicated genes. Potential use of RNA silencing technology in combination with forward genetic approaches for analyzing duplicated genes is discussed.  相似文献   

Characterization of Erysipelothrix rhusiopathiae strains isolated from recent swine erysipelas outbreaks in Japan     

To H  Sato H  Tazumi A  Tsutsumi N  Nagai S  Iwata A  Nagano T 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2012,74(7):949-953

The objective of the present study was to characterize Erysipelothrix sp. strains from recent erysipelas outbreaks in Japan. Eighty-three (100%) strains were identified as E. rhusiopathiae, based on serotyping and spaA PCR. Fifty (60.3%), 5 (6.0%), and 28 (33.7%) strains were isolated from animals with acute, subacute and chronic outbreaks, respectively, of which 79 (95.2%), 1 (1.2%), and 3 (3.6%) belonged to serotypes 1a, 2a, and untypeable, respectively. Fifteen strains (including 3, 2, and 10 from acute, subacute, and chronic cases, respectively) were sensitive to acriflavine, and showed high levels of virulence in mice; of which strains from acute cases, and from subacute and chronic cases killed 100%, and 80 to 100% mice, respectively at challenge doses of 10(2) CFU per mouse. Based on sequence analysis of a 432-bp hypervariable region in spaA gene, 83 strains could be divided into 3 groups: (i) group 1 (3 strains of serotype 1a) had Ala-195 and Ile-203; (ii) group 2 (76 strains of serotype 1a and 3 of untypeable) had Asp-195 and Met-203; and (iii) group 3 (one strain of serotype 2a) had Asn-195 and Ile-203. The results of the present study suggest that the serotype 1a strains belonging to the group 2 might be widespread in pig populations in Japan.  相似文献   

Comparative proteomic analysis of liver mitochondrial proteins derived from cloned adult pigs reconstructed with Meishan pig fibroblast cells and European pig enucleated oocytes     

Takeda K  Tasai M  Iwamoto M  Oe M  Chikuni K  Nakamura Y  Tagami T  Nirasawa K  Hanada H  Pinkert CA  Onishi A 《The Journal of reproduction and development》2012,58(2):248-253

Somatic cell nuclear transfer (SCNT) has been exploited in efforts to clone and propagate valuable animal lineages. However, in many instances, recipient oocytes are obtained from sources independent of donor cell populations. As such, influences of potential nuclear-cytoplasmic incompatibility, post SCNT, are largely unknown. In the present study, alterations in mitochondrial protein levels were investigated in adult SCNT pigs produced by microinjection of Meishan pig fetus fibroblast cells into enucleated matured oocytes (maternal Landrace genetic background). Mitochondrial fractions were prepared from liver samples by mechanical homogenization and differential centrifugation. Liver mitochondria were then subjected to two-dimensional difference gel electrophoresis (2-D DIGE). Protein expression changes were confirmed with a volume ratio greater than 2 fold (P<0.05). 2-D DIGE analysis further revealed differential expression of three proteins between the Meishan (n=3) and Landrace (n=3) breeds. Differential expression patterns of 16 proteins were detected in SCNT pig liver tissue (n=3) when compared with Meishan control samples. However, none of the 16 proteins correlated with the three differentially expressed Meishan and Landrace liver mitochondrial proteins. In summary, alteration of mitochondrial protein expression levels was observed in adult SCNT pigs that did not reflect the breed difference of the recipient oocytes. Comparative proteomic analysis represents an important tool for further studies on SCNT animals.  相似文献   

Feeding Aspergillus awamori reduces skeletal muscle protein breakdown and stimulates growth in broilers     

Ahmed A. SALEH  Yahya Z. EID  Tarek A. EBEID  Akira OHTSUKA  Masahiro YAMAMOTO  Kunioki HAYASHI 《Animal Science Journal》2012,83(8):594-598

This study was conducted to show that dietary supplementation of a fungus, Aspergillus awamori called Koji in Japan, reduces skeletal muscle protein breakdown and stimulates growth in broiler chickens. A total of 30 chicks at 15 days of age was divided into control and two treatment groups (10 birds per treatment). Control group was fed basal diet and treatment groups were fed the basal diets supplemented with A. awamori at levels of 0.05% and 0.2%. The birds were raised for 12 days from 15 to 27 days of age and then the effect on growth, organ weights and plasma 3‐methylhistidine concentration and digestibilities of protein and energy was evaluated. The messenger RNAs (mRNAs) of atrogin‐1, ubiquitin, proteasome, m‐calpain, µ‐calpain, β‐actin, myosin and pax‐7 in the breast muscle were also measured. Body weight gain and breast muscle weight were increased, although feed intake was decreased by the fungus and thus feed efficiency was increased. Protein and energy digestibilities were increased. Furthermore, plasma 3‐methylhistidine concentration was decreased by the fungus. The mRNAs of atrogin‐1, ubiquitin, proteasome, m‐calpain and µ‐calpain were all decreased. The mRNA of β‐actin but not myosin and pax‐7 was slightly increased by the fungus. In conclusion, feeding A. awamori improves growth performance because skeletal muscle proteolytic activity is reduced and digestibilities of energy and protein are increased.  相似文献   

Exterminating Sunamphitoe namhaensis using carbon dioxide in seawater for culturing brown algae Sargassum horneri seedlings     

Seta  Tomofumi  Kurashima  Akira 《Fisheries Science》2021,87(3):365-370

The amphipod Sunamphitoe namhaensis grazes on seedlings of Sargassum horneri in culture. To exterminate S. namhaensis, we immersed them in seawater with dissolved carbon dioxide—a treatment used previously to remove copepods in cultures of abalone and sea cucumber. Experiments were conducted under different CO₂ (aq) concentrations and for different lengths of time. Amphipod mortality was 100% following immersion in CO₂ seawater with a CO₂ (aq) concentration of 26,262 µmol/kg (pH 5.0) for 60 min. When algal seedlings were immersed in CO₂ seawater under these same conditions, their survival rate and growth were not affected.

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Expression of genes related to corticotropin production and glucocorticoid feedback in corticotroph adenomas of dogs with Cushing's disease     

Teshima T  Hara Y  Takekoshi S  Teramoto A  Osamura RY  Tagawa M 《Domestic animal endocrinology》2009,36(1):3-12

Cushing's disease caused by pituitary corticotroph adenoma is a common endocrine disease in dogs. A characteristic biochemical feature of corticotroph adenomas is their relative resistance to negative feedback by glucocorticoids. In this study, we examined gene expression related to adrenocorticotropic hormone (ACTH) production and secretion, and the negative feedback by glucocorticoids in canine corticotroph adenoma. We used resected corticotroph adenomas from 10 dogs with Cushing's disease. In order to investigate the alteration of gene expression between corticotroph adenoma and normal corticotrophic cells, ACTH-positive cells in the anterior lobe were microdissected using a laser-capture microdissection system, and mRNA levels of proopiomelanocortin (POMC), corticotropin releasing hormone receptor 1 (CRHR1), glucocorticoid receptor (GR), mineralocorticoid receptor (MR), and 11 beta hydroxysteroid dehydrogenase (11HSD) type 1 and type 2 were determined using real-time RT-PCR. POMC, CRHR1, and 11HSD2 mRNA levels in corticotroph adenoma were greater than those in normal corticotrophic cells (POMC, 5.5-fold; CRHR1, 4.9-fold; 11HSD2, 4.2-fold, P<0.01, respectively). MR and 11HSD1 mRNA levels in corticotroph adenoma were lower than those in normal corticotrophic cells (MR, 2.2-fold; 11HSD1, 2.9-fold, P<0.01, respectively). GR mRNA levels did not differ between corticotroph adenoma and normal corticotrophic cells. Our results may help to understand the increased ACTH production and the resistance to negative feedback suppression by glucocorticoids in canine corticotroph adenomas. These changes in gene expression may have a role in the growth of canine corticotroph adenoma, and help elucidate the pathophysiology of dogs with Cushing's disease.  相似文献   

Satellite tracking of young Steller sea lion off the coast of northern Hokkaido     

Norihisa Baba  Hiroshi Nitto and  Akira Nitta 《Fisheries Science》2000,66(1):180-181

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Surgical management of combined hydrocephalus, syringohydromyelia, and ventricular cyst in a dog     

Hasegawa T  Taura Y  Kido H  Shibazaki A  Katamoto H 《Journal of the American Animal Hospital Association》2005,41(4):267-272

Combined hydrocephalus, syringohydromyelia, and a ventricular cyst were found by magnetic resonance imaging in a 7-month-old, male miniature dachshund with gait abnormalities and altered mentation. Clinical signs did not improve with prednisone therapy. Ventriculoperitoneal shunting improved the clinical signs and anatomical abnormalities. Repeated operations were needed to replace the ventricular drainage tube at 3 and 31 months after the first surgery. The animal died suddenly with severe tonic-clonic, generalized seizures 3 weeks after the third operation. Ventriculoperitoneal shunting may be a viable treatment for syringohydromyelia associated with hydrocephalus.  相似文献   

Prevalence and characterization of Staphylococcus aureus and enterotoxigenic Staphylococcus aureus in retail raw chicken meat throughout Japan     

Kitai S  Shimizu A  Kawano J  Sato E  Nakano C  Kitagawa H  Fujio K  Matsumura K  Yasuda R  Inamoto T 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2005,67(3):269-274

A total of 444 samples of raw chicken meat (thighs, breasts, wings, livers, gizzards, hearts and ovaries) that retailed at 145 different supermarkets in 47 prefectures in Japan were examined for contamination with Staphylococcus aureus in association with its enterotoxigenicity. S. aureus was isolated from 292 (65.8%) of the samples, and from 131 of the 145 supermarkets. There was no significant difference in the detection rate of S. aureus according to the type of meat examined. About 80% of 714 isolates belonged to the poultry (57.1%) and human biotypes (22.1%). Seventy-eight (21.7%) of 360 isolates were enterotoxigenic and isolated from 78 samples in 53 supermarkets in 31 prefectures. Staphylococcal enterotoxins (SEs) produced were SEB (50 isolates), SEA (14), SEC (8), SED (2), SEA+SEB (2), and SEA+SEC (2). Most of the enterotoxigenic isolates belonged to the human and poultry biotypes, coagulase type VII, VIII or IV, and were lysed by phages of group III. Identical SE types, biotypes, coagulase types and pulsed-field gel electrophoresis (PFGE) patterns were shown in isolates from different types of meat at the same supermarket and from samples taken from different supermarkets in the same prefectures or in isolates from samples obtained from several different prefectures. Among the 50 SEB-producing isolates, 27 yielded three similar PFGE patterns that differed by only a few fragments, suggesting that they were closely related genetically. The three patterns were found in isolates of samples that retailed at 17 supermarkets in 11 prefectures, indicating that they may be disseminated among raw chicken meat in Japan.  相似文献   

Biological activity of recombinant bovine interferon tau using an Autographa californica nuclear polyhedrosis virus expression system     

Takahashi H  Inumaru S  Takahashi M  Watanabe S  Iga K  Yokomizo Y  Geshi M  Okano A  Okuda K 《The Journal of reproduction and development》2003,49(6):433-440

Bovine interferon (bIFN) tau, which plays a key role in maternal-fetal recognition of pregnancy, was expressed by an Autographa californica nuclear polyhedrosis virus expression system. cDNA coding bIFNtau was derived from cultured trophoblast cells. The recombinant (r) bIFNtau had high antiviral activity (1 x 10 (8) IU/mg) and the molecular weight of rbIFNtau was estimated to be 23 kDa by Western blotting analysis. We investigated the biological effect of rbIFNtau on prostaglandin (PG) F(2alpha) synthesis in cultured bovine endometrial epithelial cells in the presence or absence of oxytocin (OT, 100 nM). rbIFNtau suppressed basal and OT-induced PGF(2alpha) production in a dose-dependent manner (1-1,000 ng/ml). These results showed that biologically active rbIFNtau was produced in the baculovirus expression system, and that rbIFNtau had the ability to suppress the synthesis of PGF(2alpha) from bovine endometrial epithelial cells.  相似文献