An intracytoplasmic injection of deionized bovine serum albumin immediately after somatic cell nuclear transfer enhances full-term development of cloned mouse embryos

In mouse somatic cell nuclear transfer (SCNT), polyvinylpyrrolidone (PVP) is typically included in the nuclear donor injection medium. However, the cytotoxicity of PVP, which is injected into the cytoplasm of oocytes, has recently become a cause of concern. In the present study, we determined whether bovine serum albumin deionized with an ion-exchange resin treatment (d-BSA) was applicable to the nuclear donor injection medium in SCNT as an alternative to PVP. The results obtained showed that d-BSA introduced into the cytoplasm of an enucleated oocyte together with a donor nucleus significantly enhanced the rate of in vitro development of cloned embryos to the blastocyst stage compared with that of a conventional nuclear injection with PVP in SCNT. We also defined the enhancing effects of d-BSA on the blastocyst formation rate when d-BSA was injected into the cytoplasm of oocytes reconstructed using the fusion method with a hemagglutinating virus of Japan envelope before oocyte activation. Furthermore, immunofluorescence experiments revealed that the injected d-BSA increased the acetylation levels of histone H3 lysine 9 and histone H4 lysine 12 in cloned pronuclear (PN) and 2-cell embryos. The injection of d-BSA before oocyte activation also increased the production of cloned mouse offspring. These results suggested that intracytoplasmic injection of d-BSA into SCNT oocytes before oocyte activation was beneficial for enhancing the in vitro and in vivo development of mouse cloned embryos through epigenetic modifications to nuclear reprogramming.     

Iron-deficiency response and expression of genes related to iron homeostasis in poplars

Iron (Fe) deficiency is a serious agricultural problem, especially in calcareous soils, which are distributed worldwide. Poplar trees are an important biomass plant, and overcoming Fe deficiency in poplars will increase biomass productivity worldwide. The poplar Fe-deficiency response and the genes involved in poplar Fe homeostasis remain largely unknown. To identify these genes and processes, we cultivated poplar plants under Fe-deficient conditions, both in calcareous soil and hydroponically, and analyzed their growth rates, leaf Soil and Plant Analyzer Development (SPAD) values, and metal concentrations. The data clearly showed that poplars have notable growth defects in both calcareous soil and a Fe-deficient hydroponic culture. They exhibited serious chlorosis of young leaves after 3 weeks of Fe-deficient hydroponic culture. The Fe concentrations in old leaves with high SPAD values were markedly lower in Fe-deficient poplars, suggesting that poplars may have good translocation capability from old to new leaves. The Zn concentration in new leaves increased in Fe-deficient poplars. The pH of the hydroponic solution decreased in the Fe-deficient culture compared to the Fe-sufficient culture. This finding shows that poplars may be able to adjust the pH of a culture solution to better take up Fe. We also analyzed the expression of Fe homeostasis-related genes in the roots and leaves of Fe-sufficient and Fe-deficient poplars. Our results demonstrate that PtIRT1, PtNAS2, PtFRO2, PtFRO5, and PtFIT were induced in Fe-deficient roots. PtYSL2 and PtNAS4 were induced in Fe-deficient leaves. PtYSL3 was induced in both Fe-deficient leaves and roots. These genes may be involved in the Fe uptake and/or translocation mechanisms in poplars under Fe-deficient conditions. Our results will increase a better understanding of the Fe-deficiency response of poplars and hence improve the breeding of Fe-deficiency-tolerant poplars for improved biomass production, the greening of high pH soils, and combatting global warming.     

Expression analysis of defense-related genes in wild kiwifruit (Actinidia rufa) tolerant to bacterial canker

Journal of General Plant Pathology - Kiwifruit bacterial canker, which is caused by Pseudomonas syringae pv. actinidiae biovar 3 (Psa3), is found throughout kiwifruit-growing countries. Here, we...     

Ingestion of medium chain fatty acids by lactating dairy cows increases concentrations of plasma ghrelin

The purpose of this study was to elucidate the effects of medium-chain fatty acids (MCFAs) on plasma ghrelin concentration in lactating dairy cows. Five early-lactating Holstein cows were randomly assigned to 2 dietary treatments in a crossover design with 2-wk periods. Treatments consisted of diets supplemented or not (control) with calcium salts of MCFAs (MCFA-Ca; 1.5% dry matter). Plasma hormone and metabolite concentrations in blood samples taken from the jugular vein were measured on the morning of feeding on day 14 of each period. Dry matter intake, milk protein, and lactose content of cows fed the MCFA-Ca diet were decreased compared with controls, but with no change in milk yield. Plasma ghrelin concentrations were higher in cows fed the MCFA-Ca diet; however, no significant effect was found on glucagon-like peptide-1 concentrations in plasma. Plasma insulin concentrations decreased, but plasma glucagon concentrations remained unchanged in cows fed the MCFA-Ca diet. The concentrations of nonesterified FAs, total cholesterol, and β-hydroxybutyrate in plasma increased in these cows. In conclusion, dietary MCFAs increase the plasma ghrelin concentrations in lactating dairy cows.     

Motility and fertility of boar semen after liquid preservation at 5°C for more than 2 weeks

This study investigated the effects of skim milk on the quality and fertility of boar spermatozoa under long‐term chilled preservation. Semen samples were stored in Modena solution supplemented with 0 (control) to 50 mg/mL skim milk at 5°C for 4 weeks; spermatozoa stored with 7.5 and 15 mg/mL of skim milk (7.5‐SM and 15‐SM groups, respectively) exhibited significantly higher motility indices than those of the control group up to 3 weeks (P < 0.05), and the 7.5‐SM group showed improved motility indices even after 4 weeks (P < 0.05). In vitro fertilization using spermatozoa in the 7.5‐SM and 15‐SM groups stored at 5°C for 2 weeks showed significantly higher fertilization rates of spermatozoa and the development rates to blastocyst than the control group (P < 0.05), and the 7.5‐SM group showed similar rates of fertilization and blastocyst formation in the fresh non‐stored spermatozoa group. After artificial insemination using spermatozoa stored for 2 weeks in the 7.5‐SM group, healthy piglets were obtained. Boar spermatozoa can be stored at 5°C in a Modena solution containing skim milk. Supplementation of 7.5 mg/mL skim milk improves boar spermatozoa motility and fertility even after liquid preservation at 5°C for 2 weeks.     

Investigation of Soft Wheat Flour Quality Factors Associated with Sponge Cake Sensory Tenderness

Relationships among soft wheat quality parameters relating to sponge cake volume and sensory tenderness were investigated. Sixteen soft wheats from the 2008–2009 crop and 11 from the 2009–2010 crop, including Japanese soft wheat cultivars, advanced breeders' lines, and western white wheat imported from the United States, were milled and evaluated for protein content, sucrose solvent retention capacity value, specific surface area, flour pasting properties, batter pasting viscosity, sodium dodecyl sulfate sedimentation (SDSS) volume, farinograph properties, specific cake volume, and sensory tenderness score to investigate their relationships. Batter pasting viscosity was measured with a Rapid Visco Analyzer (RVA) at 2 min after reaching 90°C in heating a mixture with equal weights of flour, sucrose, and water. RVA minimum viscosity of flour suspension in water was the most influencing factor and positively correlated to specific cake volume, and RVA batter pasting viscosity and SDSS volume were negatively correlated. Meanwhile, protein content and SDSS volume were strongly negatively correlated with sensory tenderness score. Stepwise multiple regression analysis selected protein content and specific cake volume as independent variables to predict sensory tenderness score; however, SDSS volume and farinograph properties relating to protein strength were not selected. Protein content affected sponge cake tenderness independently of specific cake volume, which was related to differences in cake density.     

Characterization of a novel potyvirus tentatively named <Emphasis Type="Italic">Ornithogalum</Emphasis> virus 3, successfully isolated from <Emphasis Type="Italic">O. thyrsoides</Emphasis> co-infected with two other potyviruses by single-aphid inoculation

A potyvirus tentatively named Ornithogalum virus 3 (OV-3) was successfully isolated by single-aphid transmissions from O. thyrsoides mix-infected with OV-3, Ornithogalum mosaic virus (OrMV) and Ornithogalum stripe mosaic virus (OrSMV). OV-3, a flexuous, rod-shaped particle of ca. 690 nm, was sap and aphid transmissible. The virus had a narrow host range and caused necrotic mosaic on O. thyrsoides under cold conditions. We therefore propose the name Ornithogalum necrotic mosaic virus (OrNMV) for OV-3. A synergistic increase in symptom severity was apparent on O. thyrsoides mix-infected with OrSMV/OrNMV, but not with either OrMV/OrNMV or OrMV/OrSMV. The nucleotide sequence data reported is available in the DDBJ/EMBL/GenBank databases under accession number AB282754.     

Molecular species of collagen in muscular and vertebral parts of white sturgeon Acipenser transmontanus

The molecular species of collagen in the muscular and vertebral parts of white sturgeon Acipenser transmontanus were examined by biochemical techniques. Pepsin-solubilized collagens were prepared from these parts, and fractionated into major and minor collagen fractions by differential salt precipitation at acidic or neutral pH. Collagens contained in these fractions showed several features characteristic of fish type I and V collagens, respectively, in SDS-PAGE patterns and the amino acid compositions of both parts. These results suggest that at least two molecular species, corresponding to type I and V collagens, are distributed in the muscular and vertebral parts of white sturgeon.