Reliable enzyme-linked immunosorbent assay for the determination of soybean proteins in processed foods

Among allergenic foods, soybean is known as a food causing adverse reactions in allergenic patients. To clarify the validity of labeling, the specific and sensitive detection method for the analysis of the soybean protein would be necessary. The p34 protein, originally characterized to be p34 as an oil-body associated protein in soybean, has been identified as one of the major allergenic proteins and named Gly m Bd 30K. A novel sandwich enzyme-linked immunosorbent assay (ELISA) for the detection and quantification of the soybean protein in processed foods was developed using polyclonal antibodies raised against p34 as a soybean marker protein and the specific extraction buffer for extract. The developed sandwich ELISA method was highly specific for the soybean protein. The limit of detection (LOD) and the limit of quantification (LOQ) of the developed ELISA were 0.47 ng/mL (equivalent to 0.19 microg/g in foods) and 0.94 ng/mL (equivalent to 0.38 microg/g in foods), respectively. The recovery ranged from 87.7 to 98.7%, whereas the intra- and interassay coefficients of variation were less than 4.2 and 7.5%, respectively. This study showed that the developed ELISA method is a specific, precise, and reliable tool for the quantitative analysis of the soybean protein in processed foods.     

Effect of plant density on growth and yield of new soybean genotypes grown under early planting condition in southwestern Japan

Soybeans planted in early to mid-June (early) are less affected by rainfall during rainy season than those conventionally planted in early to mid-July in southwestern Japan. Also, narrow row cultivation is expected to increase soybean yield and save labor for inter-tillage and ridging. Field experiments were performed in 2014 and 2015 to test the effect of plant density (high, middle, and low) under early planting condition on growth, yield, and several agronomical traits of Sachiyutaka A1 and three new genotypes (Sakukei 155, Kanto 127 and Shikoku 15). Early planting was performed in mid- to late June, even though rainy season started in early June. Higher plant densities produced 13% greater yield than low plant density through an increase in biomass accumulation, especially at R5. Among yield components, only pods m^?2 was significantly and positively correlated with yield, indicating that an increase in pods m^?2 led to a greater yield with higher plant densities. The yields of Sachiyutaka A1 were relatively stable for two years, but the lodging resistance should be further improved. Shikoku 15 had greater yield potential and lodging resistance, but its resistance to damping-off disease should be improved. Sakukei 155 with medium plant density produced relatively high and stable yield with less lodging. Although the yield of Kanto 127 fluctuated between experimental years, this genotype showed higher yield potential in higher plant densities with less lodging in 2015. Thus, Sakukei 155 and Kanto 127 with high or medium density may be suitable for early planting in this region.     

Control mechanisms for producing antimicrobial factors in ruminant mammary gland

Mastitis, a symptom of inflammation in mammary tissue by infection with various kinds of bacteria, causes huge economic losses in the milk industry. One of the popular methods for treatment of mastitis is antibiotics, although this prohibits milk shipping and sometimes causes resistant microbes. Therefore, a new strategy to treat mastitis without antibiotics is eagerly required around the world. Antimicrobial factors belong to innate immunity and can start their function extremely early after bacterial stimulation. These factors have antimicrobial activity for a broad spectrum of bacteria. Elucidation of causal mechanisms and functions of antimicrobial factors in the mammary gland is thought to result in suitable methods for prevention and treatment of mastitis. Therefore, this review introduces traits of some antimicrobial factors and the mechanisms for expressing, producing and secreting them in the mammary gland. For antimicrobial factors, lingual antimicrobial peptide (LAP), S100A7, cathelicidin and lactoferrin are controlled in different sites and different time courses, suggesting that antimicrobial factors play different roles for local defense against bacterial infection in the mammary gland. These findings will contribute to the development of prevention and treatment methods for mastitis.     

Effect of Steel Slag to Improve Soil Quality of Tsunami-Impacted Land while Reducing the Risk of Heavy Metal Bioaccumulation

After the tsunami caused by the Great East Japan Earthquake, marine sediment was taken from the sea bottom and deposited over local agricultural fields. The marine sediment already contained an unknown amount of heavy metals, due to anthropogenic activities prior to the tsunami, which might affect plants, animals, and humans. Furthermore, soil salinity in tsunami-inundated land greatly increased. Three different amounts of steel slag were employed as pretreatment agent in order to improve agricultural soil quality. The soil samples treated with 2% of steel slag present a remarkable increase of A. thaliana biomass production with low BCF and TF values for most of the heavy metals. It was concluded that steel slag pretreatment used in the tsunami-inundated agricultural lands produced a noteworthy improvement in soil quality which lead to a positive stimulative effect on plant growth, and the slag addition treatment proved to be a promising treatment that might be used for phytostabilization of slightly contaminated soils.     

Three-dimensional structural analysis of mitochondria composing each subtype of fast-twitch muscle fibers in chicken

In a previous study, the three-dimensional structures of mitochondria in type I and type IIb muscle fibers of chicken were analyzed. The study reported differences in the shape of the mitochondria and the distribution of lipid droplets. In this study, we three-dimensionally analyzed mitochondria and lipid droplets of type II muscle fiber subtypes IIa, IIb, and IIc of chicken lateral iliotibial muscle in the same field of view using correlative light electron microscopy (CLEM) and array tomography methods. The reconstructed images showed that the mitochondria of type IIa muscle fiber were thick and aligned along the myofibrils, and many lipid droplets were embedded in the mitochondria. The mitochondria of type IIb muscle fibers were intermittent, aligned along the myofibrils, and showed contact between adjacent horizontal mitochondria. No lipid droplets were observed in type IIb muscle fiber. In type IIc muscle fiber, we observed irregularly shaped mitochondria with small diameters aligned along the myofibrils. Lipid droplets not only were embedded in the mitochondria but also existed independently in some cases. The combination of array tomography and CLEM methods enabled three-dimensional electron microscopic observation of mitochondria in different subtypes of type II muscle fibers. The subtypes of type II muscle fibers differed in mitochondrial occupancy and morphology and in lipid droplet distribution, and characteristics that had been demonstrated biochemically were also demonstrated ultrastructurally.     

一组新的小麦EST-SSR标记开发及其在遗传图谱构建中的应用

为了丰富小麦功能分子标记的数量,为小麦功能基因的定位、比较基因组学、小麦起源和进化等方面的研究奠定基础。从前期利用小麦转录组数据鉴定出的8 389个EST-SSR序列中,随机选取585对引物得分大于95的标记进行分析,以川麦42和川农16杂交后自交获得的重组自交系群体为试验材料,利用新开发出的EST-SSR标记,并结合SSR标记和SRAP标记,构建了一张遗传图谱。结果表明,585对EST-SSR引物中有555对能在亲本川麦42和川农16中扩增出稳定清晰的条带,引物有效性为94.87%。其中有44对EST-SSR引物在川麦42和川农16中表现出明显的、稳定的多态性,多态性率为7.93%。所构建的遗传图谱中含有本次新开发的EST-SSR标记的连锁群共12个,由163个标记组成,包括17个EST-SSR标记,73个SSR标记和73个SRAP标记,覆盖小麦基因组长度1 551.5 cM,相邻标记之间的平均距离为13.11 cM。本研究丰富了小麦功能分子标记的数量,为小麦功能基因的定位、比较基因组学、小麦起源和进化等方面的研究奠定基础。