Ecological pre-release risk assessment of two genetically engineered, bioluminescent Rhizobium meliloti strains in soil column model systems

In order to identify potential ecological risks associated with the environmental release of two Rhizobium meliloti strains, genetically engineered with the firefly-derived luciferase gene (luc), a pre-release greenhouse investigation was conducted. The upper 4 cm of soil columns (30 cm diameter; 65 cm depth), which were filled according to the horizons of an agricultural field (loamy sand), were inoculated with seeds of Medicago sativa (alfalfa) and R. meliloti cells at approximately 5×10⁶ cells·g^–1 soil. Four treatments were tested: inoculation with a non-engineered wild type strain (2011), strain L33 (luc ⁺), strain L1(luc ⁺, recA^–) and non-inoculated controls. The fate of the engineered strains was followed by two methods: (1) selective cultivation and subsequent detection of bioluminescent colonies and (2) PCR detection of the luc gene in DNA, directly extracted from soil. Strain R. meliloti L33 declined to 9.0×10⁴ cfu·g^–1 soil within 24 weeks and to 2.8×10³ cfu·g^–1 soil within 85 weeks in the upper 25 cm of the soil columns. Decline rates for R. meliloti L1 were not significantly different. Vertical distribution analysis of the recombinant cells after 37 weeks revealed that in three of four columns tested, the majority of cells (>98%) remained above 10 cm soil depth and no recombinant cells occurred below 20 cm depth. However, in one column all horizons below 20 cm were colonized with 2.2×10⁴ to 6.8×10⁴ cfu g^–1 soil. Ecological monitoring parameters included organic substance, total nitrogen, ammonium and nitrate, microbial biomass, culturable bacteria on four different growth media and the immediate utilization of 95 carbon sources (BiologGN) by soil-extracted microbial consortia. None of the parameters was specifically affected by the genetically engineered cells. Received: 6 December 1996     

Effectiveness of dicyandiamide as a nitrification inhibitor in biochar-amended soil

Overuse of nitrogen (N) fertilizers may lead to many environmental issues via N leaching into groundwater and agricultural runoff into surface water. Biochar, a sustainable soil amendment agent, has been widely studied because of its potential to retain moisture and nutrients. However, recent studies have shown that biochar has a very limited ability to improve the retention of negatively charged nitrite (NO₂^-) or nitrate (NO₃^-). Although positively charged ammonium (NH₄⁺) can be better held by biochar, it is usually susceptible to nitrification and can be easily transformed into highly mobile NO₂^-and/or NO₃^-. In practice, dicyandiamide (DCD) has been used to inhibit nitrification, preserving N in its relatively immobile form as NH₄⁺. Therefore, it is likely that the effects of DCD and biochar in soils would be synergistic. In this study, the influences of biochar on the effectiveness of DCD as a nitrification inhibitor in a biochar-amended soil were investigated by combining the experimental results of incubation, adsorption isotherm, and column transport with the simulated results of different mathematical models. Biochar was found to stimulate the degradation of DCD, as the maximum degradation rate slightly increased from 1.237 to 1.276 mg kg^-1 d^-1 but the half-saturation coefficient significantly increased from 5.766 to 9.834 mg kg^-1. Considering the fact that the availability of DCD for nitrification inhibition was continuously decreasing because of its degradation, a novel model assuming non-competitive inhibition was developed to simulate nitrification in the presence of a decreasing amount of DCD. Depending on the environmental conditions, if the degradation of DCD and NH₄⁺ in biochar-amended soil is not significant, improved contact due to the mitigated spatial separation between NH₄⁺ and DCD could possibly enhance the effectiveness of DCD.     

INVESTIGATION ABOUT DETERMINATION OF COLUMN EFFECTIVE LENGTH OF STEEL FRAMES WITH FLEXIBLE JOINTS

Moment slope displacement equations of beams with two end semirigid connection restraints are covered. The evaluation of column effective length factor both braced frames and unbraced frames with flexible joints is derived. Determination of column effective length method proposed herein is. exactly the same as those of rigid frames which was well established in Chineses Specification of Steel Structures, however, the restraining stiffness of girders to columns should be amended semirigid connection is applied.     

Experimental Investigation of "Column with Weaker Beam" Frame and "Column with Equally Strong Beam" Frame

By the way of experiment and analysis of two "one story and one bay" reinforced concrete frames, namely a "beam with weaker column" frame and a "column with equally strong beam" frame, the features of internal force redistribution are investigated and an elementary comparison about bending moment redistribution of three kinds of frames namely the "column with weaker beam", "beam with weaker column" and "column with equally strong beam" is given. Through this comparison, the objective existence of such "three kinds of frame" is proven, the moment redistribution is different for each frame and for the moment modulation, the elastic moment diagram and the difference of the internal force redistribution in different frames should be considered together.     

免疫亲合柱在小麦真菌毒素检测中的应用及毒素污染分析

利用真菌毒素特异性抗体制备的免疫亲合柱净化样品，检测了31个小麦样品中5种真菌毒素的污染情况。结果显示，31个样品污染毒素比较普遍，但污染水平低，小麦样品处于安全状态。具体情况是：AFB1，阳性率为74．19％，平均值为1．13μg／kg，中值为0．3μg／kg，最大值为11μg／kg；OTA，阳性率为100％，平均值为1．59μg／kg，中值为1μg／kg，最大值为3μg／kg；ZON，阳性率为100％，平均值为83μg／kg，中值为71μg／kg，最大值为330μg／kg；T-2，阳性率为22．58％，平均值为49μg／kg，中值为0，最大值为500μg／kg；DON，阳性率为6．45％，平均值为200μg／kg，中值为0，最大值为3800μg／kg。     

Thin-layer chromatographic analysis of flower color phenotypes in Dendranthema grandiflorum Ramatuelle inbreds and clonal cultivars

Summary Four glasshouse and 13 garden chrysanthemum cultivars or inbred seedlings from the color groups white, yellow, pink/purple, and orange/red/bronze were analyzed with thin-layer chromatography (TLC) for qualitative and quantitative pigment constituents. TLC confirmed the presence of eight carotenoids in yellow and bronze cultivars. Qualitative differences exised only for spots 5–8. Rf values for lutein matched those of spot 4. Flavonoid analysis of purples and bronzes revealed two anthocyanidins, one of which was cyanidin, one flavone, and two phenylpropanoids. White cultivars had four phenylpropanoids, two of which were caffeic and ferulic acids. In most cases, inbred and non-inbred cultivars within a given phenotypic class did not exhibit qualitative differences. Color analysis could be pursued at any level of inbreeding, but inheritance studies would be severely hampered using non-inbred genotypes of this hexaploid floricultural crop.     

Herbicidal Potential and Quantification of Suspected Allelochemicals from Four Grass Crop Extracts

Unknown compounds in crop plants are inhibitory to seed germination and early seedling growth of weed plants. A Petri dish assay showed that barley (Hordeum vulgare L.), oats (Avena sativa L.), rice (Oryza sativa L.) and wheat (Triticum aestivum L.) extracts significantly reduced root growth of alfalfa (Medicago sativa L.), barnyard grass (Echinochloa crus‐galli, Beauv. var. oryzicola Ohwi.) and eclipta (Eclipta prostrata L.). As the concentration of crop extracts increased, root growth of the test plants were significantly reduced. A high‐performance liquid chromatography analysis with nine standard phenolic compounds showed that the concentrations and compositions of allelopathic compounds depend on the extracted plant extracts. Caffeic acid, hydro‐cinnamic acid, ferulic acid, m‐coumaric acid, p‐coumaric acid and coumarin were present in all the crop plant species, and hydro‐cinnamic acid were detected as the highest amount. Coumarin at 10^?3 m significantly inhibited root growth of alfalfa and barnyard grass more than that of eclipta. The research suggests that extracts of barley, oats, rice and wheat have an allelopathic effect on alfalfa, barnyard grass and eclipta and that the findings of bioassay were considerably correlated with the type and amount of causative allelochemicals, indicating that the allelopathic effects on three test plants were ranked in order of wheat (highest), barley, rice and oats (lowest). The results may have value in enabling weed control based on natural plant extracts or crop residues in the fields.     

α-Tomatine and other steroidal glycoalkaloids in fruits of tomato lines resistant to the glasshouse whitefly (Trialeurodes vaporariorum Westw.)

Summary The total steroidal alkaloid compositions of the mature-harvested fruits of the tomato cultivar Allround (Lycopersicon esculentum), of the species L. hirsutum glabratum, resistant to the glasshouse whitefly (Trialeurodes vaporariorum), and of four resistant lines were determined by capillary gas chromatography. The contents of -tomatine in the red-ripe fruits of Allround and in those of the lines were low (5 mg/kg fresh weight). The green-mature fruits of the wild species had a high content (3390 mg/kg fresh weight) of -tomatine. The total alkaloid profiles of Allround and of the lines were similar. Comparison of flame ionisation detector response with thermoionic detector response indicated that in addition to tomatidine, the aglycone of -tomatine, a small amount of one other steroidal alkaloid might be present in Allround and in the lines. The wild species contained five compounds which possibly are steroidal alkaloids.The safety level of -tomatine in tomatoes is discussed. It was concluded that, in respect of the glycoalkaloids of the mature-harvested fruits, the resistant tomato breeding lines are as safe for human cosumption as the standard cultivar.     

Determination of Phytic Acid in Cottonseed Kernels by Ion Chromatography

Cottonseed meal is an important plant protein resource of feed industry and breeding industry, although it contains many antinutritional factors. Phytic acid, as one of the mainly antinutritional factor, cannot be digested easily by monogastric animals, such as fowls and pigs, consequently results in the decreased bio-availability of material elements and leads to the environment pollution by the phytate in excreta. Thus far, study of breeding cottons containing low phytic acid gradually becomes the hotspot, and the method of fully extracting phytic acid from cotton seeds and precise measurement is becoming more and more important and necessary. A new method for the rapid extraction and determination of phytic acid in cottonseed kernels by ion chromatography was developed in this study. After the diethyl ether treatment of dry cottonseed kernel powder, the samples were suspended in 10ml 0.66 mol·L^－1 HCl and boiled for 1 hour. Then the samples were shaken at 4 ℃ for 12 hours and centrifuged. The supernatant was diluted and purified for loading. To analyze the sample, DIONEX ICS-2000 ion chromatography, AG11-HC Guard (4 mm×50 mm) protection column and AG11-HC Guard (4 mm×250 mm) separation column were used in controlled conditions (washing buffer: 30.0 mmol·L^－1 KOH; ream speed: 1.0 mL·min^－1; collecting time: 25 min; 75 mA current; temperature: 30 ℃; loading amount: 20 μL). The results showed that R2 of linear regression is 0.999, which means the regression relation is great. In addition, the standard deviations of all the indexes are within 5% and the average recovery rate is 98.64%-102.31%. The detection limit, showing the minimum detectable concentration of analyte in the sample, is 0.059 μg·mL^－1, and the quantitation limit, showing the minimum amount of analyte in a sample can be quantitatively determined, is 0.196 μg·mL^－1. This method is easy and accurate with great reproducibility, showing a strong application value.