Near-isogenic lines of durum wheat: their development and plant characteristics

Summary The effect of specific plant characteristics on the grain and biomass yield of durum wheat can be accurately determined by using isogenic lines, which, however, were not usually available. This study reported the effects of long glume, glaucousness, glume pubescence, black glume and purple culm on the yield and its associated characteristics in near-isogenic lines of durum wheat cv. LD222, which were developed by continuous backcrossing. The long glume trait which resulted in a large photosynthetic area did not enhance yield. Increased glume size associated with the P gene tended to increase the main culm dominance, characterized such as vigorous main culm and weak tillers, and plant height, but to reduce tillering and spike number. Consequently, grain yield and harvest index declined. Under the adequate water supplying condition of the present study, the glaucous trait was beneficial for grain yield. The trait of glume pubescence did not excert any significant effect on the yield related characteristics in the LD222 background. The traits of black glume and purple culm reduced the number of spikes per unit area and the number of kernels per unit area.     

Construction and characterization of a BAC library of soybean

We have constructed a soybean (Glycine max (L.) Merrill) bacterial artificial chromosome (BAC) library from green leaf protoplasts of the cultivar, Misuzudaizu. The library contains 53,760 clones with an average insert size of 116 kb. About 2.9% chloroplast DNA origin was revealed by PCR and colony hybridization. Apart from 2.8% clones having no insert, this library represents 5.2 genome equivalents. With this genome coverage, the probability of having any DNA sequence represented in the library is higher than 99.5%. Three-dimensional pools of the BAC library in combination with the use of a high efficiency genome scanning (HEGS) electrophoresis facilitate rapid and efficient PCR-based screening. An average of five positive clones were identified after screening the BAC library with SSR and STS markers. BAC-end walking was performed for three SSR associated BACs. This library will provide a good resource for positional cloning of agronomically and biologically important QTL genes that Misuzudaizu possesses.     

Purification and characterization of a corrinoid compound from Chlorella tablets as an algal health food

Vitamin B(12) content of an algal health food, Chlorella tablets (Chlorella sp.), was determined by both Lactobacillus leichmannii ATCC 7830 microbiological and intrinsic factor-chemiluminescence methods. The values of 200.9-211.6 microg/100 g dry weight determined by the chemiluminescence method were similar to the values (201.3-285.7 microg/100 g dry weight) determined by the microbiological method. A corrinoid compound was purified to homogeneity from the Chlorella tablets and characterized. The purified corrinoid compound was identified as vitamin B12, on the basis of silica gel 60 TLC, C18 reversed-phase HPLC, 1H NMR spectroscopy, and UV-Vis spectroscopy.     

Investigation of the Heavy Metal Sources in Relation to Automobiles

In order to investigate the sources of roadside heavy metal pollution, auto tire, gasoline, gas oil, kerosene, paint for road markings (white, yellow and red), the soot from the discharge spout, auto tire rubber, asphalt pavement, gray paint and anticorrosive on the guardrail were gathered together and analyzed. As a consequence, Pb may be originally from yellow and red road markings, and gray paint or anticorrosive. Hg is mainly emitted from the combustion of premium gasoline and diesel soot is an important source of As, Ni and Zn. Cd and Zn are mainly emitted from the abrasion of tire rubber. The wear of asphalt pavement is considered to be a source of Ni and V.     

Synthesis of haptens for development of antibodies to alkylphenols and evaluation and optimization of a selected antibody for ELISA development

The development of an enzyme-linked immunosorbent assay (ELISA) based on polyclonal antibodies for a class of endocrine disrupting compounds, 4-nonylphenol, is described. The parent molecule was derivatized at the ortho position of the free phenolic hydroxyl group to obtain the hapten, NP1, and it was conjugated with keyhole limpet hemocyanin, which was used as an immunogen. Four antisera were generated and screened against three coating antigens. The most sensitive ELISA from the screening tests (antiserum NP03As, 1/1000, and coating antigen NP1-BSA, 1 microg/mL) was further optimized and characterized. The influence of various physicochemical factors (organic solvent, pH, ion strength) was investigated. Methanol as the additive organic solvent was found to be the best organic solvent for the ELISA, with optimal sensitivity observed at a concentration of 5%. The ELISA parameters were changed at more acidic or basic pH values, whereas higher ionic strengths strongly suppressed the I(50) value and the maximum absorbance. The most sensitive ELISA for 4-nonylphenol exhibited an I(50) value of 38.6 +/- 5.5 microg/L, with a dynamic range from 12 to 350 microg/L, and the lower limit of detection was 7.7 +/- 1.3 microg/L. The optimized ELISA displayed no significant cross-reaction against the parent compounds, nonylphenol ethoxylates, degradation products, carboxylates, and bisphenol A, except in 4-octylphenol.     

Development of an enzyme-linked immunosorbent assay for the pyrethroid cypermethrin

A competitive enzyme-linked immunosorbent assay (ELISA) for the detection of cypermethrin was developed. Two haptens, the trans- and cis-isomers of 3-[(+/-)-cyano-3-(2,2-dichloroethenyl)-2,2-dimethylcyclopropanecarbonyloxy]methyl]phenoxyacetic acid, were conjugated with thyroglobulin as immunogens. Four antisera were generated and screened against six different coating antigens. The assay that was the most sensitive for cypermethrin was optimized and characterized. The IC(50) for cypermethrin was 13.5 +/- 4.3 microg/L, and the lower detection limit (LDL) was 1.3 +/- 0.5 microg/L. This ELISA had relatively low cross-reactivities with other major pyrethroids, such as deltamethrin, phenothrin, resmethrin, fluvalinate, and permethrin. Methanol was found to be the best organic cosolvent for this ELISA, with an optimal sensitivity observed at a concentration of 40% (v/v). The assay parameters were unchanged at pH values between 5.0 and 8.0, whereas higher ionic strengths strongly suppressed the absorbances. To increase the sensitivity of the overall method, a C(18) sorbent-based solid-phase extraction was applied to various domestic and environmental water samples. The water samples, fortified with cypermethrin, were analyzed according to this method. Good recoveries and correlation with spike levels were observed.     

Inactivation of staphylococcal enterotoxin-A with an electrolyzed anodic solution.

Electrolyzed anodic NaCl solutions [EW+], prepared by the electrolysis of 0.1% NaCl, have been shown to instantly inactivate most pathogens that cause food-borne disease. Elimination of food-borne pathogens does not necessarily guarantee food safety because enterotoxins produced by pathogens may remain active. We have tested whether EW+ can inactivate Staphylococcal enterotoxin A (SEA), one of the major enterotoxins responsible for food poisoning. Fixed quantities of SEA were mixed with increasing molar ratios of EW+, and SEA was evaluated by reversed-phase passive latex agglutination (RPLA) test, immunoassay, native polyacrylamide gel electrophoresis (PAGE), and amino acid analysis after 30 min incubations. Exposure of 70 ng, or 2.6 pmol, of SEA in 25 microL of PBS to a 10-fold volume of EW+, or ca. 64.6 x 10(3)-fold molar excess of HOCl in EW+, caused a loss of immuno-reactivity between SEA and a specific anti-SEA antibody. Native PAGE indicated that EW+ caused fragmentation of SEA, and amino acid analysis indicated a loss in amino acid content, in particular Met, Tyr, Ile, Asn, and Asp. Staphylococcal enterotoxin-A excreted into culture broth was also inactivated by exposure to an excess molar ratio of EW+. Thus, EW+ may be a useful management tool to ensure food hygiene by food processing industries.     

Evaluation of alternative substrates for determining methane-oxidizing activities and methanotrophic populations in soils

The magnitude of methane emission is a net result of methane production and the oxidation rate. The possibility of measuring oxidized products of alternative substrates of methane monooxygenase was examined to determine methane-oxidizing ability of soils, and to count methanotrophic populations in soils. Wetland rice soils were incubated under methane containing air to enirch the methanotrophs. Methane loss and oxygen uptake were inhibited by acetylene, dimethylether, and nitrapyrin (N-Serve). Acetylene was used routinely, because it inhibited methane oxidation even at a low concentration of 0.03 to 0.06 l ml^-1 in the incubation headspace. Propylene at 10 kPa was used as an alternative substrate of methane monooxygenase, and the formation of propylene oxide was measured. When soils were incubated under methane, their methane-oxidizing activity increased. Propylene oxide formation increased simultaneously. Acetylene also blocked propylene oxidation. The results of several experiments and propylene oxide formation (r=0.87 after long-transformation). These results indicate that propylene oxide formation can be used as a semiquantitative measure of the methane-oxidizing activity of soils. The colonies of soluble methane monooxygenase-forming methanotrophs were counted on Cu-deficient methanotroph agar medium by the formation of naphthol from haphthalene. The counts increased from 10⁴ (0 days) to 10⁷ (21 days) g^-1 soil during oxic incubation under methane.