Development of a screening method for genetically modified soybean by plasmid-based quantitative competitive polymerase chain reaction

A novel type of quantitative competitive polymerase chain reaction (QC-PCR) system for the detection and quantification of the Roundup Ready soybean (RRS) was developed. This system was designed based on the advantage of a fully validated real-time PCR method used for the quantification of RRS in Japan. A plasmid was constructed as a competitor plasmid for the detection and quantification of genetically modified soy, RRS. The plasmid contained the construct-specific sequence of RRS and the taxon-specific sequence of lectin1 (Le1), and both had 21 bp oligonucleotide insertion in the sequences. The plasmid DNA was used as a reference molecule instead of ground seeds, which enabled us to precisely and stably adjust the copy number of targets. The present study demonstrated that the novel plasmid-based QC-PCR method could be a simple and feasible alternative to the real-time PCR method used for the quantification of genetically modified organism contents.     

基于烧失量指数评估滨海沉积物中有机质特征

Understanding organic matter characteristics(e.g., amount and state) of sediments is necessary for evaluating both the sediment and water environments; however, methods that have been proposed to investigate these characteristics are relatively expensive. This study attempted to evaluate the organic matter characteristics of littoral sediments by employing solely the loss on ignition(LOI)method, which is the most economical and accessible method in developing countries. Different types of sediments were first oven dried at 100℃ and then continuously burned at 200, 300, 400, 500, and 600℃ for 4 h at each temperature. The mass LOI at each temperature was calculated and compared with the 100℃ oven-dried weight. Our results suggested that the mass LOI across the temperature range of 200–300℃(LOI_(200-300)):mass LOI at 600℃(LOI_(600)) ratio(LOI_(200-300)/LOI_(600)) could represent different types of sediments as well as the organic matter:organic C ratio. Compared with the sediments unaffected by wastewater, the sediments that were more influenced by the inflow of domestic wastewater had a higher LOI_(200-300)/LOI_(600). Interestingly, LOI_(200-300)/LOI_(600) could also represent changes in the organic matter characteristics as a function of organic matter decomposition in sediments. In conclusion, the organic matter characteristics of littoral sediments could be evaluated using solely the LOI method, particularly with LOI_(200-300)/LOI_(600).     

Characterizing nicotianamine aminotransferase: Improving its assay system and details of the regulation of its activity by Fe nutrition status

Under iron deficient conditions, graminaceous plants secrete mugineic acid family phytosiderophores (MAs) from their roots to dissolve sparingly soluble iron compounds in the rhizosphere, and take up iron in the form of an Fe³⁺-MAs complex (Takagi 1976). A good correlation has been reported between the tolerance of Fe-deficiency and the amount of secreted MAs (Takagi 1993). Therefore, by using the genes involved in MAs biosynthesis, molecular breeding might produce transgenic plants tolerant to Fe-deficiency with a high level of MAs secretion. The biosynthetic pathway of MAs from L-methionine has been clarified (Fig. 1) and the enzymes participating in this process are now being investigated to isolate the genes responsible. Nicotianamine aminotransferase (NAAT) catalyzes the amino group transfer between nicotianamine (NA) and 2-oxoglutaric acid (Fig. 1). In order to purify NAAT, enzyme assay methods for NAAT have been developed and modified (Shojima et al. 1990; Ohata et al. 1993; Kanazawa et al. 1994). Some characteristics of NAAT have been reported using these enzyme assay methods (Kanazawa et al. 1994, 1995). Here, we further investigate some characteristics of this enzyme to improve the enzyme assay method, namely 1) the effect of K⁺ and Mg²⁺ on NAAT activity in vitro, and 2) the direct influence of MAs, Fe³⁺, and Fe²⁺ on NAAT activity. In addition, based on these results, the induction of enzyme activity by Fe-deficiency and suppression of the activity by Fe-resupply was investigated, by applying the new enzyme assay method.     

Underreporting of energy intake among Japanese women aged 18-20 years and its association with reported nutrient and food group intakes

OBJECTIVES: To evaluate the ratio of energy intake to basal metabolic rate (EI/BMR) among young female Japanese adults, and to compare the lifestyle and dietary characteristics between relatively low and high reporters. DESIGN: Dietary intakes were assessed over a 1-month period with a validated, self-administered, diet history questionnaire, and lifestyle variables were assessed by a second questionnaire designed for this survey. The ratio of EI/BMR was calculated from reported energy intake and estimated basal metabolic rate. SUBJECTS: In total, 1889 female Japanese university students aged 18-20 years who were enrolled in dietetics courses. RESULTS: Ninety-five per cent of the subjects were classified into a non-obese group (body mass index (BMI) <25 kg m(-2); mean+/-standard deviation (SD): 20.8+/-2.6 kg m(-2)). EI/BMR was 1.43+/-0.40 (mean+/-SD). Sixty-eight per cent of the subjects showed an EI/BMR level below the possibly balanced value of 1.56, 37% showed EI/BMR below the minimum survival value of 1.27 and 2% of the subjects showed EI/BMR exceeding the maximum value for a sustainable lifestyle of 2.4. BMI, body weight and BMR decreased significantly with the increase in EI/BMR (P<0.001). The percentage of energy from carbohydrate was significantly higher, whereas those from fat and protein were significantly lower, among the lower EI/BMR groups. As for food groups, a significantly declining trend from the lowest to the highest EI/BMR groups was observed for cereals. CONCLUSION: Underreporting, rather than overreporting, of energy intake was predominant in this relatively lean Japanese female population. BMI was the most important factor affecting the reporting accuracy of energy intake.     

Time course study of aluminum-induced callose formation in barley roots as observed by digital microscopy and low-vacuum scanning electron microscopy

To clarify the mechanism(s) involved in the short-term inhibition of root elongation by AI, we monitored the morphological changes of barley roots by digital microscopy. Within 30 min after exposure to 37 µM AI, the surface of the root epidermis in the region of a distance of 1.5 mm from the root tip became rough and began to show signs of damage. After 38 min, callose was rapidly excreted from the junction between the root cap and the root epidermis, and formed a spherical lump approximately 60 µm in diameter. The fine structure of the callose deposits on the root surface was analyzed by low-vacuum scanning electron microscopy. After 50 min, there was a significant increase in the callose contents in the distal 0.6 mm part. At the same time, root elongation stopped completely. Fluorescence staining indicated that callose was localized on the surface of the cell elongation area (the elongation zone of primary roots and root hairs), but not on the surface of the meristem. The root growth reduction associated with AI treatment may be due to the use of sugar substrates for callose formation instead of cellulose formation.     

Effects of Inoculation of Phototrophic Purple Bacteria on Grain Yield of Rice and Nitrogenase Activity of Paddy Soil in a Pot Experiment

Phototrophic purple bacteria (PPB) are one group of dinitrogen (N₂)-fixing bacteria often found in the anoxic and photic zone of paddy soil. In the present study, the effects of inoculation of PPB on grain yield of rice and changes in their populations and nitrogenase activity were investigated with and without surface application of rice straw in a pot experiment. As an inoculant, Rhodopseudomonas palustris strain KN122 isolated from a paddy soil was selected, and the cell suspension was inoculated into the floodwater once or three times during the cultivation. As a result, the inoculation of PPB increased the grain yield of rice. Compared to the control, the grain yield was 9% higher in the inoculated plot without rice straw application. The treatment was more effective in combination with rice straw application. In the plots where PPB were inoculated with rice straw, grain yields were 21% (single inoculation) and 29% (triple inoculation) higher. Populations of PPB in the plots without rice straw application significantly increased by the inoculation, unlike those in the plots with rice straw application, except for the bulk soil. Phototrophic and heterotrophic nitrogenase activities (acetylene-reducing activity) associated with the soils or residues of the rice straw were not affected by PPB inoculation throughout the experiment. This indicates that the inoculation of PPB into floodwater may not be effective for enhancing N₂ fixation in paddy soils and that the beneficial effect of PPB inoculation on the grain yield of rice may be due to unidentified functions of PPB other than biological N₂ fixation.     

Barium isotopes in allende meteorite: evidence against an extinct superheavy element

Carbon and chromite fractions from the Allende meteorite that contain isotopically anomalous xenon-131 to xenon-136 (carbonaceous chondrite fission or CCF xenon) at up to 5 x 10(11) atoms per gram show no detectable isotopic anomalies in barium-130 to barium-138. This rules out the possibility that the CCF xenon was formed by in situ fission of an extinct superheavy element. Apparently the CCF xenon and its carbonaceous carrier are relics from stellar nucleosynthesis.     

Hemorrhagic necrotizing splenitis in a slaughter pig infected with Arcanobacterium species

A 6-month-old barrow presented with lethargy, inappetence and dysstasia. At necropsy, multiple coalescing hemorrhagic foci were detected in the margins of the spleen. Gram-positive bacilli were isolated from the spleen, kidney, muscle and liver. Comparative 16S rDNA gene sequencing analysis of the isolates (TO16177) revealed that they would be the same species of unpublished Arcanobacterium species strain HJ57-14E (accession no. gi 18873551) (99.7% similarity based on a comparison of 675 bp). Histologic examination of the splenic tissue sections revealed extensive necrosis and inflammation, and gram-positive bacilli were discernible. Multifocal necrosis was also detected in the liver. Immunohistochemically, the isolates were cross-reacted with polyclonal antibodies against Arcanobacterium pyogenes and Actinomyces naeslundii, and the reaction was strong for the latter. Similar reactions were found in the suppurative lesions of the tonsil, and occasionally in the spleen and lymph nodes. The present results indicate that the unpublished Arcanobacterium species induced multiple organ failure accompanied by acute hemorrhagic necrotizing splenitis in this growing-finishing pig.