Development of avian embryo manipulation techniques and their application to germ cell manipulation

The development of chicken embryo culture techniques, from single‐cell stage to hatching, makes it possible to manipulate developing embryos at any developmental stage. Production of germline chimeric chickens by the transfer of stage X blastodermal cells or primordial germ cells enables the manipulation of germline cells in vitro. Production of transgenic chickens has been attempted by the retroviral vector method, microinjection of DNA into a fertilized ovum at the single‐cell stage, use of chimeric intermediates produced by the transfer of stage X blastodermal cells or primordial germ cells, manipulation of spermatozoa, and in vivo manipulation of gonads. So far, the only non‐viral method that has successfully produced transgenic chickens is microinjection of DNA into a fertilized ovum. Manipulation of primordial germ cells could become an efficient system for producing transgenic chickens by combining it with the highly efficient transfection method or the in vitro culture system for primordial germ cells. Preservation of avian genetic resources has now become possible by cryopreservation of stage X blastodermal cells or primordial germ cells as well as spermatozoa. The development of nuclear transfer techniques for avian species is necessary.     

Allele‐specific polymerase chain reaction typing and sequencing of mitochondrial D‐loop region in broiler chickens in Japan

This study aimed to comprehend a feature of single nucleotide polymorphism (SNP) in mitochondrial DNA (mtDNA) mainly of general broiler chickens in Japan. We typed two SNP sites (199C/T and 792A/G) of the D‐loop region in mtDNA by allele‐specific PCR (AS‐PCR) in 359 broiler (182 chunky and 177 cobb) and 506 layer (233 White Leghorn, 140 Barred Plymouth Rock and 133 Rhode Island Red) chickens. The SNP of 199C or 792A by AS‐PCR was observed in the chunky and cobb chickens, and not in the layers. The haplotype 199T/792G was observed in a part of cobb and all layers. By the result of AS‐PCR haplotyping and the broiler brands, the D‐loop region was sequenced in 44 broiler chickens (20 chunky and 24 cobb) and compared with the layers’ sequence data. Among the broiler and layer chickens, 21 SNP sites (including one insertion) and 11 sequence haplotypes were observed. Haplotype variation or correspondence was observed in and between the broiler brands. This study provides important information to establish a chicken meat traceability system by SNP haplotyping of mtDNA in Japan.     

Irradiation history of Itokawa regolith material deduced from noble gases in the Hayabusa samples

Noble gas isotopes were measured in three rocky grains from asteroid Itokawa to elucidate a history of irradiation from cosmic rays and solar wind on its surface. Large amounts of solar helium (He), neon (Ne), and argon (Ar) trapped in various depths in the grains were observed, which can be explained by multiple implantations of solar wind particles into the grains, combined with preferential He loss caused by frictional wear of space-weathered rims on the grains. Short residence time of less than 8 million years was implied for the grains by an estimate on cosmic-ray-produced (21)Ne. Our results suggest that Itokawa is continuously losing its surface materials into space at a rate of tens of centimeters per million years. The lifetime of Itokawa should be much shorter than the age of our solar system.     

Isolation and characterization of a cDNA from Catharanthus roseus which is highly homologous with phosphate transporter

The PIT1 gene which is highly homologous with phosphate transporter was isolated from Catharanthus roseus and analyzed. The cBNA PIT1 contained an open reading frame of 542 amino acids and its sequence showed a 31, 30, and 34% identity with the phosphate transporter of Saccharomyces cerevisiae (PBO84), Neurospora crassa (PHO-5), and Glomus versiforme (GvPT), respectively. Furthermore, the cDNA PIT1 encoded a highly hydrophobic protein with 12 putative membrane-spanning regions and contained a conserved amino acid sequence reported in the human glucose transporter super-family* S. cerevisiae strain DpU (pho84 knockout strain) was unable to grow on low phosphate (55 μM) medium (LP medium). Expression of the PIT1 cDNA enabled DpU to grow on LP medium. Northern hybridization analysis revealed that the PIT1 gene was expressed in roots, stems, and young whole plant of C. roseus, but not in leaves.     

Moisture distribution of soaked rice grains observed by magnetic resonance imaging and physicochemical properties of cooked rice grains

Moisture distribution in individual polished rice grains was observed during soaking by magnetic resonance (MR) imaging, and a nuclear magnetic resonance (NMR) signal intensity profile (SI-profile) was generated from the MR image. The water penetration pattern during soaking roughly showed dissimilar trends between different varieties of japonica and japonica-indica hybrid rice. NMR signal intensity at the completion of water absorption varied within each grain; high at the periphery and the central region and low in the area between the periphery and the central region. High moisture content within the central region is due to loosely packed starch granules. The SI-profile was congruent for grains of the same variety harvested in different regions and years and characterized a grain moisture distribution for each variety. Moisture distribution was compared using SI-profiles for varieties with different amylose contents and new varieties bred for specific end-uses in Japan. The NMR signal intensity, which is related to the moisture content, at the surface of soaked grain was negatively correlated to the grain amylose content. The NMR signal intensities at the surface of soaked grain negatively correlated with the overall hardness of the cooked rice grain as measured by the single-grain low-high compression test.     

Overexpression of the LASAP2 gene for secretory acid phosphatase in white lupin improves the phosphorus uptake and growth of tobacco plants

Secretion of acid phosphatase (APase) from the roots to take up phosphorus (P) is a well-known strategy of plants under P-deficient conditions. White lupin, which shows vigorous growth in low-P soils, is noted for its ability to secrete APase under P-deficient conditions. The APase secreted by white lupin roots is stable in soil solution and shows low substrate specificity, suggesting that genetic modification of plants using the APase gene LASAP2 might improve their ability to use organic P. The objective of the present study was to evaluate the potential of LASAP2 transgenic plants to increase organic P utilization. Dry matter production and P accumulation were higher in LASAP2 transgenic tobacco plants grown in gel media containing soluble phytate as the sole P source than in wild-type tobacco plants. Phosphorus uptake by the transgenic plants also increased in soil culture conditions. LASAP2 was apparently more effective in the liberation of organic P, including phytate, in the soil than the native tobacco APase. Thus, the enzymatic stability of LASAP2 in the soil appears to be an important factor for P acquisition.     

Metabolite profiling of spinach (Spinacia oleracea L.) leaves by altering the ratio of in the culture solution

To observe the effect of different forms of N, we have applied metabolic profiling using gas chromatography mass spectrometry to evaluate the metabolite composition of spinach. The aim of the present study was to find an appropriate indicator of overall metabolic response to N source. The effect of the     ratio on spinach tissue was investigated, comparing two cultivars that differed in their ability to use N. There was wide variation in     absorption without any distinct growth inhibition in either cultivar. Statistical analysis revealed that the metabolites could be broadly divided into two types, correlating either positively or negatively with     uptake in the culture solution. Principal component analysis (PCA) was an effective tool that extracted the indexes of the effect of the ratio of N forms as PCA scores. Close relationships between metabolite profiles and     uptake were observed, indicating that metabolic profiling is able to distinguish the invisible metabolic change in mature leaves of spinach and may provide a new tool for the investigation of quality features or rhizosphere nutritional condition.     

Efficiency of soil and water conservation practices in different agro-ecological environments in the Upper Blue Nile Basin of Ethiopia

In developing countries such as Ethiopia, research to develop and promote soil and water conservation practices rarely addressed regional diversity. Using a water-balance approach in this study, we used runoff plots from three sites, each representing a different agro-ecological environment, e.g., high, mid and low in both elevation and rainfall, in the Upper Blue Nile Basin of Ethiopia to examine the runoff response and runoff conservation efficiency of a range of different soil and water conservation measures and their impacts on soil moisture. The plots at each site represented common land use types(cultivated vs. non-agricultural land use types) and slopes(gentle and steep). Seasonal runoff from control plots in the highlands ranged 214–560 versus 253–475 mm at midlands and 119–200 mm at lowlands. The three soil and water conservation techniques applied in cultivated land increased runoff conservation efficiency by 32% to 51%, depending on the site. At the moist subtropical site in a highland region, soil and water conservation increased soil moisture enough to potentially cause waterlogging, which was absent at the lowrainfall sites. Soil bunds combined with Vetiveria zizanioides grass in cultivated land and short trenches in grassland conserved the most runoff(51% and 55%, respectively). Runoff responses showed high spatial variation within and between land use types, causing high variation in soil and water conservation efficiency. Our results highlight the need to understand the role of the agro-ecological environment in the success of soil and water conservation measures to control runoff and hydrological dynamics. This understanding will support policy development to promote the adoption of suitable techniques that can be tested at other locations with similar soil, climatic, and topographic conditions.