Production of mouse fibroblast growth factor 4 in E. coli and its application for isolation and maintenance of mouse trophoblast stem cells in vitro

Fibroblast growth factor 4 (FGF4) promotes isolation of trophoblast stem (TS) cells from mouse blastocysts and maintenance of TS cells in an undifferentiated state in vitro. To date, commercially available, bacterially expressed human FGF4 (RhFGF4) has been used generally for this purpose. In this study, HismFGF4, a 6x histidine-tagged mouse FGF4, was produced in E. coli and purified using heparin column chromatography. We demonstrated that HismFGF4 (25 ng/ml) more efficiently generates mouse TS cells from a single blastocyst than RhFGF4 (25 ng/ml) and that TS cells isolated and maintained with HismFGF4 retained their ability to differentiate into the trophoblast cell lineage in vitro. In addition, TS cells cultured with HismFGF4 (25 ng/ml) were maintained in an undifferentiated state better than with RhFGF4 (25 ng/ml). To the best of our knowledge, this is the first application of a mouse FGF4 derivative for isolation and maintenance of mouse TS cells.     

Effect of the expression level of an AGAMOUS-like gene on the petaloidy of stamens in the double-flowered lily, ‘Elodie’

Double-flowered lilies, in which stamens are converted to petaloid organs, are valuable for horticulturists. ‘Elodie’ is a double-flowered lily cultivar in which stamens are homeotically converted into petaloid organs in whorl 3. The ‘Elodie’ cultivar shows individual variation in stamen structure and it was therefore classified into the following three types according to the strength of petaloidy of the stamens: weak (type-I), intermediate (type-II), and strong (type-III) phenotypes. The AGAMOUS (AG) gene is a class C floral identity gene in Arabidopsis thaliana that is involved in the formation of stamens and carpels. An AG-like gene was isolated from ‘Elodie’ (LelAG1) and its expression was compared between flower types. The LelAG1 gene was expressed in whorls 3 and 4, but not in whorls 1 and 2 in all flower types. In type-I flowers, LelAG1 was expressed strongly in whorls 3 and 4, while its expression was significantly decreased in whorl 3 in type-III flowers. In type-II intermediate phenotype flowers, the expression level of LelAG1 in whorl 3 was reduced by 60%. These results suggest that the expression level of AG-like genes is correlated with the degree of petaloidy of the stamens.     

Flowering Date1, a major photoperiod sensitivity gene in adzuki bean,is a soybean floral repressor E1 ortholog

Adzuki bean is an important legume crop originating in temperate regions, with photoperiod in sensitivity being a key factor in its latitudinal adaptation. The Flowering Date1 (FD1) gene has a large effect on the photoperiodic response of flowering time, but the molecular basis for the effect of this locus is undetermined. The present study delimited the FD1 locus to a 17.1 kb sequence, containing a single gene, an E1 ortholog (VaE1). A comparison between Vigna angularis ‘Shumari’ (photoperiod insensitive) and ‘Acc2265’ (photoperiod sensitive) identified 29 insertions/deletions and 178 SNPs upstream of VaE1 in the FD1 locus. VaE1 expression in ‘Acc2265’ was greater under long-day than short-day conditions, whereas VaE1 expression in ‘Shumari’ was lower regardless of day length. These findings suggested that responsible gene of FD1 is a VaE1, which acts as a floral repressor by being upregulated in response to long-day conditions. The inability to upregulate VaE1 under long-day conditions was linked to its ability to flower under these conditions. These results provide greater understanding of the molecular control of a flowering date and clues enabling the breeding of adzuki bean at higher latitudes.     

Greenhouse gases emission from paddy soil during the fallow season with and without winter flooding in central Japan

Many papers on measurements of greenhouse gases (GHGs) emission in rice paddies during a rice cropping season have been published. During a non-cropping season between Nov. and Apr., we investigated direct and indirect GHGs emissions in rice paddies. The indirect GHGs emission was evaluated as the amount of dissolved gases leaching from the paddy fields. Water management practices for the experiment were (1) continuous flooding (CF) and (2) non-flooding (NF). Although the direct CO₂ emission in the CF treatment was remained nearly zero during the non-cropping period, direct CO₂ emission in the NF treatment was continuously observed throughout the non-cropping period. The concentration of dissolved N₂O in the NF treatment was below the detection limit of the instrument during the non-cropping period except immediately after the flooding and before the drainage. The concentration of dissolved N₂O kept approximately 2 µg L^?1 during the non-cropping period in the CF treatment. The direct CH₄ emission and dissolved CH₄ were not observed during the non-cropping period. Total gas emission in the NF treatment was 10 times as large as that in the CF treatment. Direct CO₂ emission accounted for more than 90 % of the total emission in both treatments.     

Teratoma of the umbilical cord in a giraffe (Giraffa camelopardalis reticulata)

A 12-year-old pregnant female giraffe (Giraffa camelopardalis reticulata) died approximately 2 months prior to her anticipated parturition date. At necropsy, a mass measuring approximately 20 x 36 x 20 cm was observed, attached to the umbilical cord, the latter being otherwise normal in appearance. Histologically, the mass contained 3 germinal tissue components with areas of squamous epithelium, respiratory epithelium, primitive neural tissues, glial tissue, peripheral nerve, adipose tissue, cartilage, and smooth muscle. Based on these findings, the tumor was diagnosed as a teratoma originating from the umbilical cord. This is possibly the second reported case of umbilical cord teratoma in animals.     

Computed tomographic images of discospondylitis in a calf

A 2-month-old male Japanese Black calf was presented with a 30-day history of progressive ataxia. Antemortem examination using computed tomography (CT) revealed narrowing of the disc spaces due to destruction of intervertebral structures between the first and second thoracic vertebrae and between the second and third thoracic vertebrae. Osteolysis was evident as irregular hypoattenuating lesions within the opposing end plates of the first, second and third thoracic vertebrae. Pseudomonas aeruginosa was detected as the causative bacteria, and discospondylitis was diagnosed. To the best of our knowledge, this is the first bovine case report describing the application of CT for the diagnosis of discospondylitis.     

Use of soil nonexchangeable potassium by paddy rice with clay structural changes under long-term fertilizer management

To prove the hypothesis that paddy rice utilizes soil nonexchangeable potassium (neK) and causes associated structural changes in clay minerals, K status and clay mineralogy of 22 surface soils from three paddy fields under long-term fertilizer management for 51–93 years were investigated. Soil neK content was determined as the difference between 1 mol L⁻¹ hot HNO₃ extractable K and 1 mol L⁻¹ ammonium acetate exchangeable K. Clay mineralogy was identified by X-ray diffraction (XRD). The radiocesium interception potential (RIP), an index of frayed edge sites in the interlayer sites of 2:1 type clay minerals, was also determined. The neK contents under the -K and NPK treatments were considerably lower than those under the unfertilized treatment in all the fields, indicating the exploitation of soil neK by rice. XRD analysis of the clay samples revealed 7% shift from the 1.0 peak to 1.4 nm one under the -K treatment compared with the unfertilized one, and the amounts of neK were negatively correlated with those of RIP (p < .01), suggesting the expansion of interlayer spaces of the 2:1 type phyllosilicates such as mica due to the release of neK. In addition, the neK content positively correlated with K balance of the long-term experiments (p < .05). The differences of neK between unfertilized K and -K treatments corresponded to 22–157 kg K ha⁻¹, or 0.42–1.68 kg K ha⁻¹ year⁻¹. In conclusion, utilization of considerable amount of soil neK under K depleted conditions should be considered to establish sustainable K management for paddy rice.     

Impact of farmland reclamation on soil distribution in Japan: the case of Andosols in Nagano Prefecture

Journal of Soils and Sediments - In modern times, farmlands in Japan have been reconstructed at a large-scale through the cutting and filling of soils. In principle, such reconstruction should be...     

Glutathione levels in phloem sap of rice plants under sulfur deficient conditions

Glutathione which is an abundant reduced sulfur compound in plants is considered to play important roles in the transmission of the sulfur nutrient status between organs within the plant body and in the long-distance transport of reduced sulfur. We determined the concentrations of glutathione, γ-glutamylcysteine (γ-EC), sulfate in the rice phloem sap collected by the insect laser technique. Phloem sap was collected from the plants cultured in sulfur-deficient and control solutions. The concentration of glutathione in the rice phloem sap was higher than that of sulfate in both control and sulfur-deficient plants. Under sulfur-deficient condition, the concentration of glutathione in the phloem sap did not decrease, whereas the sulfate concentration decreased significantly. The pattern of changes in γ-EC concentration was similar to that of glutathione. These data indicate the presence of mechanisms for the maintenance of a constant glutathione concentration in the phloem sap in rice plants under sulfur deficiency, whereas the sulfate concentration was found to be relatively unstable.