Xanthan gum and locust bean gum substrate improves bovine embryo development

One of the major difference between the in vivo and in vitro embryonic environments is the stiffness of the culture substrate. Xanthan gum (XG) and locust bean gum (LBG) are natural materials that are safe, inexpensive and easy to handle. In this study, we investigated the effects of using a polysaccharide culture substrate made from 1% XG and 1% LBG (XG-LBG gel) on bovine embryonic development. Oocytes collected from bovine ovaries were subjected to maturation, and fertilization to generate embryos at an early developmental stage (>4 cell stage). Cleaved embryos were further cultured in a well of 96-well cell culture plate coated with or without XG-LBG gel for 5 days. While the developmental rate up to the blastocyst stage did not differ between the two culture systems (control, 38.0 vs. gel, 38.6%), blastocysts developed on the XG-LBG gel produced significantly high cell numbers and ATP content. Embryos cultured on XG-LBG gels for 24 hr had high expression levels of F-actin and a highly even distribution of E-cadherin. In addition, embryos developed on XG-LBG gel demonstrated increased translocation of YAP to the nucleus and increased connective tissue growth factor (CTGF) protein levels (downstream of Hippo signalling). These findings suggest that soft culture substrates improve embryonic development by enhancing mechanotransduction, including YAP-CTGF signalling.     

The assessment of genetic diversity within and among the eight subpopulations of Japanese Black cattle using 52 microsatellite markers

Japanese Black cattle are at risk for genetic homogeneity due to intensive use of a few sires. Therefore, assessment of the actual genetic diversity of this breed is important for future breeding plans. In the present study, we investigated the genetic diversity within and among eight subpopulations of Japanese Black cattle using 52 microsatellite markers. The parameters for genetic diversity of Japanese Black cattle were comparable to those of other cattle breeds, suggesting that the relatively high genetic diversity of the breed. However, upon comparison among the eight subpopulations, the Hyogo subpopulation showed markedly low genetic diversity. The results of the pairwise F_ST values, phylogenetic network and structure analysis indicated that the Hyogo population has remarkably high level of genetic differentiation from other populations, while Yamagata, Niigata, Hiroshima and Kagawa populations have low levels of genetic differentiation. Furthermore, multidimensional scaling plots indicated that individuals in some subpopulations were separated from individuals in the other subpopulations. We conclude that while the overall genetic diversity of Japanese Black cattle is still maintained at a relatively high level, that of a particular subpopulation is significantly reduced, and therefore the effective population size of the breed needs to be controlled by correct mating strategies.     

Prediction of water resources as snow storage under climate change in the Tedori River basin of Japan

We investigate the effect of climate change on water resources as snow storage (WRSS) using a regional climate model with a 20-km mesh (MRI-RCM20), in the Tedori River basin of Japan. We make four main predictions. First, annual average temperature in Kanazawa is predicted to increase by 2.7 °C by 2081–2100, compared to the period 1981–2000. The smallest predicted temperature difference is 1.1 °C in August, and the largest is 3.4 °C in April and December. The annual average temperature is predicted to change from 14.5 to 17.2 °C. Second, annual average precipitation is predicted to increase by 3.8 %, and precipitation during the snowfall season (December–February) is predicted to decrease by 13.2 %, as a result of increasing temperatures. Third, average WRSS for the period 2001–2100 is predicted by degree-day and tank model analyses. Both the analyses show very similar results. The degree-day analysis shows 200 mm of average WRSS in 2100, and 477 mm in 2050. These values constitute 26 and 63 % of the WRSS value in 2000 (754 mm). The tank model analysis shows WRSS of 260 mm in 2100 and 523 mm in 2050, which are 33 and 67 % of the WRSS (785 mm) in 2000, respectively. Fourth, to examine the effect on irrigation water especially on the paddy paddling period, discharge is predicted and considered under the predicted average WRSS is 520 mm in 2050, 258 mm in 2100, and reaches a minimum of 41 mm in 2100.     

Allelic frequencies and association with carcass traits of six genes in local subpopulations of Japanese Black cattle

Marker‐assisted selection (MAS) is expected to accelerate the genetic improvement of Japanese Black cattle. However, verification of the effects of the genes for MAS in different subpopulations is required prior to the application of MAS. In this study, we investigated the allelic frequencies and genotypic effects for carcass traits of six genes, which can be used in MAS, in eight local subpopulations. These genes are SCD, FASN and SREBP1, which are associated with the fatty acid composition of meat, and NCAPG, MC1R and F11, which are associated with carcass weight, coat color and blood coagulation abnormality, respectively. The frequencies of desirable alleles of SCD and FASN were relatively high and that of NCAPG was relatively low, and NCAPG was significantly associated with several carcass traits, including carcass weight. The proportions of genotypic variance explained by NCAPG to phenotypic variance were 4.83 for carcass weight. We thus confirmed that NCAPG is a useful marker for selection of carcass traits in these subpopulations. In addition, we found that the desirable alleles of six genes showed no negative effects on carcass traits. Therefore, selection using these genes to improve target traits should not have negative impacts on carcass traits.     

Metabolism of glucose,fructose, and sucrose in tobacco leaves during maturity

Phosphorus sorption and desorption processes in selected soils were investigated to evaluate the usefulness of phosphorus sorption isotherms in the estimation of the phosphorus supplying capacity of soils. There was a distinct hysteresis in the phosphorus sorption and desorption isotherms, resulting in an overestimation of the replenishing ability of soils to supply phosphorus to the soil solution, when phosphorus sorption isotherms were used for the estimations.

To attain a value of 0.2 ppm P in the soil solution, 115 µmol P/g soil are required in the Kuromatsunai Ando soil (Ochric Andosols) based on the estimation by the P desorption isotherms; this value is 29% higher than that indicated by the P sorption isotherm. Also 42 µmol P/g soil are required in the Mikatagahara Yellow soil (Helvic Acrisols)—which corresponds to a value 23% higher than that indicated by the P sorption isotherm.

Two types of hysteresis subloops were observed: A reversible type and an irreversible type. The former was found typically in the highly weathered Mikatahagara Yellow soil while the latter was observed in the Kuromatsunai Ando soil.     

Cloning of the feline GADD45 cDNA and analysis of its mutation in feline lymphoma cell lines

Growth arrest and DNA damage-inducible protein 45 (GADD45) plays an important role in suppressing multistep carcinogenesis. In this report, we describe the isolation of the complete wild-type feline GADD45 cDNA from feline tissues. Expression of feline GADD45 mRNA was detected in the liver, spleen, kidney, lung, and testis. The predicted amino acid sequences encoded by the full-length feline GADD45 cDNA display sequence homology with those from other vertebrates, and as in the case of human GADD45, cell growth suppression was observed by ectopic expression of feline GADD45. However, no mutations were detected by sequence analysis of feline GADD45 in several feline lymphoma cell lines, indicating that the GADD45 mutation might be uncommon in feline oncogenesis.     

有机水稻杂草控制技术的研究

有效控制水田杂草的发生是有机水稻栽培关键技术。本田杂草控制技术的研究采用深水管理、施用米糠以及田面覆盖稻草、稻壳等有机物的方法进行水田杂草控制试验。结果表明,深水灌溉能够有效控制水田稗草的发生;施用米糠杂草控制效果可达到药剂控制的85%左右;田面覆盖稻草、稻壳等有机物对水田杂草均有不同程度的控制效果,可降低施用米糠成本。     

Abnormal morphology in megalopae of the swimming crab,Portunus trituberculatus during seed production: causes and prevention

The occurrence of morphologically immature megalopae, which retain zoeal features such as dorsal spines and furcae of telson, is closely correlated with larval mass mortality during seed production of the swimming crab Portunus trituberculatus in Japanese hatcheries. To determine the cause of immature megalopal morphology, zoeae were reared with various supplementary schedules and density of diets (rotifer, Artemia and phytoplanktons including Chlorella vulgaris and Nannochloropsis oculata). In addition, to assess the relationship between immature morphology and endocrine control, the effect of causative dietary factor was compared with the effect of eyestalk ablation at various timing during zoeal development. Megalopal immature morphology was more distinct in Chlorella‐supplemented groups than in Nannochloropsis‐supplemented groups. High density Chlorella supplementation was associated with the highest incidence of immaturity and resulted in larval mass mortality. The premoult of the third zoeal stage was identified as a critical period at which Chlorella supplementation led to the highest incidence of immaturity. This critical period coincided with the critical period at which larval metamorphosis was regulated by the eyestalk neurosecretory system. Our results suggested that the occurrence of immature megalopal morphology under culture conditions is most likely caused by phytoplankton (especially, Chlorella) supplementation, which disrupts the endocrine regulation. On the basis of our results, we successfully prevented the occurrence of immature megalopal morphology in 500 L tanks by excluding the influence of phytoplankton before the critical period (i.e. discontinuing phytoplankton supplementation and supplying rotifer cultured with non‐phytoplankton materials).     

Basic studies on hybrid wheat breeding using the 1BL-1RS translocation chromosome / Aegilops kotschyi cytoplasm system 1. Development of male sterile and maintainer lines with discovery of a new fertility-restorer

The Aegilops kotschyi cytoplasm and a 1BL-1RS translocation chromosome that consists of the long arm of wheat chromosome 1B and the short arm of rye chromosome 1R were transferred to six spring common wheat cultivars by repeated backcrossing. Resistance to leaf rust race 21B conditioned by the Lr26 gene and a secalin subunit encoded by the Sec-1 gene, both on the 1RS arm, were used as the selection markers of the translocation chromosome. Five of the six cultivars used were converted to complete male steriles, whereas the remaining one, cv. Kitamiharu 48, retained normal fertility, after transfer of both the 1BL-1RS chromosome and Ae. Kotschyi cytoplasm. Conventional gene analysis suggested that Kitamiharu 48 carries an incompletely dominant fertility-restoring gene. The F₁ hybrids between the male steriles and ordinary common wheat cultivars recovered fertility only at a low level, indicating that a single dose of the Rfv1 gene on the 1BS arm of wheat is insufficient for full fertility restoration under spring-sowing condition. Our results are in clear contrast to complete fertility restoration under fall-sowing condition reported by Nonaka et al. (1993). Combination of the 1BL-1RS chromosome / Ae. Kotschyi cytoplasm system with a new fertility-restoring gene discovered in Kitamiharu 48 may provide a breakthrough for spring-type hybrid wheat. This revised version was published online in August 2006 with corrections to the Cover Date.     

Cellular kinetics of villous epithelial cells and m cells in rabbit small intestine

The cellular kinetics of villous columnar epithelial cells and M cells in the rabbit small intestine were determined by the use of 5-bromo-2'-deoxyuridine (BrdU) as a tracer. To identify M cells, vimentin antibody was used. The BrdU-labeled nuclei of columnar epithelial cells reached the base of intestinal villi in all portions at 1 day after BrdU administration. Thereafter, BrdU-labeled cells migrated toward the villous tip, but they did not move at a uniform speed. The epithelial cells which existed in intestinal villi on circular folds moved faster than those on mucosa other than circular folds. At 7 days after BrdU administration, the leading edge of BrdU-labeled epithelial cells already disappeared from the villous tip in all portions of the small intestine. In the ileal Peyer's patch, the BrdU-labeled nuclei of microvillous epithelial cells and vimentin-positive M cells appeared near the intestinal crypt orifice at 1 day after BrdU administration, and then migrated toward the luminal surface of the follicle-associated epithelium (FAE). As they moved toward the upper portion of FAE, the number of BrdU-labeled M cells on the side of the dome decreased simultaneously. The leading edge of BrdU-labeled epithelial cells disappeared from the top of the FAE within 7 days. These results suggest that M cells may differentiate from the undifferentiated cells in intestinal crypts within 1 day and disappear from the top of the FAE after the change of their form from M cells into microvillous epithelial cells.