Fate of the Cry1Ab protein from Bt-maize MON810 silage in biogas production facilities

Biogas plants fuelled with renewable sources of energy are a sustainable means for power generation. In areas with high infestation levels with the European corn borer, Ostrinia nubilalis (Hbn.), it is likely that transgenic Bt-maize will be fed into agricultural biogas plants. The fate of the entomotoxic protein Cry1Ab from MON810 maize was therefore investigated in silage and biogas production-related materials in the utilization chains of two farm-scale biogas plants. The Cry1Ab content in silage exhibited no clear-cut pattern of decrease over the experimental time of 4 months. Mean content for silage was 1878 +/- 713 ng Cry1Ab g(-1). After fermentation in the biogas plants, the Cry1Ab content declined to trace amounts of around 3.5 ng g(-1) in the effluents. The limit of detection of the employed ELISA test corresponded to 0.75 ng Cry1Ab g(-1) sample material. Assays with larvae of O. nubilalis showed no bioactivity of the reactor effluents. The utilization of this residual material as fertilizer in agriculture is therefore deemed to be ecotoxicologically harmless.     

Degradation of Cry1Ab protein from genetically modified maize in the bovine gastrointestinal tract

Immunoblotting assays using commercial antibodies were established to investigate the unexpected persistence of the immunoactive Cry1Ab protein in the bovine gastrointestinal tract (GIT) previously suggested by enzyme-linked immunosorbent assay (ELISA). Samples of two different feeding experiments in cattle were analyzed with both ELISA and immunoblotting methods. Whereas results obtained by ELISA suggested that the concentration of the Cry1Ab protein increased during the GIT passage, the immunoblotting assays revealed a significant degradation of the protein in the bovine GIT. Samples showing a positive signal in the ELISA consisted of fragmented Cry1Ab protein of approximately 17 and 34 kDa size. Two independent sets of gastrointestinal samples revealed the apparent discrepancy between the results obtained by ELISA and immunoblotting, suggesting that the antibody used in the ELISA reacts with fragmented yet immunoactive epitopes of the Cry1Ab protein. It was concluded that Cry1Ab protein is degraded during digestion in cattle. To avoid misinterpretation, samples tested positive for Cry1Ab protein by ELISA should be reassessed by another technique.     

Detection of recombinant DNA segments introduced to genetically modified maize (Zea mays)

Polymerase Chain Reaction (PCR) techniques are increasingly used for the detection of genetically modified (GM) crops in foods. In this paper, recombinant DNAs introduced into the seven lines of GM maize, such as Event 176, Bt11, T25, MON810, GA21, DLL25, and MON802, are sequenced. On the basis of the obtained sequence, 14 primer pairs for the detection of the segments, such as promoter, terminator regions, and construct genes, were designed. To confirm the specificities of the designed primer pairs, PCR was performed on genomic DNAs extracted from GM and non-GM maize, GM and non-GM soy, and other cereal crops. Because the presence of the corresponding DNA segments was specifically detected in GM crops by the designed primer pairs, it was concluded that this method is useful for fast and easy screening of GM crops including unauthorized ones.     

A single nucleotide polymorphism (SNP839) in the adh1 reference gene affects the quantitation of genetically modified maize (Zea mays L.)

The real-time PCR methods recommended in the European Union for the quantitation of genetically modified (GM) maize events NK603, GA21, and MON 863 measure the number of copies of the GM event in relation to those of the maize-specific adh1 reference gene. The study reported here revealed that the targeted 70 base pair adh1 region exhibits a single nucleotide polymorphism (SNP839) that hampers the binding of the reverse primer used in the adh1 detection method. Partial fragments of the adh1-A and adh1-F allele were cloned. By allele-specific real-time PCR, it was shown that SNP839 corresponds to a common allelic polymorphism in maize. As a result, the quantitation of the GM maize events mentioned is positively or negatively biased, depending on the adh1 genotype of sample and calibrant. Therefore, it is proposed to revise the quantitative detection methods for NK603, GA21, and MON 863 maize.     

Response of maize (Zea mays L.) to potassium nano-silica application under drought stress

Abstract

To investigate the influence of potassium nano-silica (PNS) on maize plant under drought stress including non-stress (NS), moderate drought stress (MDS) and severe drought stress (SDS), a factorial experiment was conducted with completely randomized blocks with three replications. Drought stress decreased the concentrations in the shoot of phosphor (P), calcium (Ca), iron (Fe), zinc (Zn), manganese (Mn) and silica (Si) and nitrogen (N), P, Ca, Fe, Zn, copper (Cu), Mn and Si concentrations of seed. There was an increase in the concentration in the N seed and shoot potassium (K) concentration under drought stress. It was observed that applying PNS increased nutrient absorption. The highest concentration of N in the seed was obtained at 100?ppm PNS. The highest concentrations of seed K and N, Cu, Mn and Si in the shoot were found when 200?ppm of PNS was applied. Applying PNS had no significant effect on the concentrations of P, Ca, sodium (Na) and Cu in the seed, and of Ca and Na in the shoot. These findings demonstrate that the application of PNS can limit the negative effects of drought stress and improve plant’s resistance against drought stress.     

First application of a microsphere-based immunoassay to the detection of genetically modified organisms (GMOs): quantification of Cry1Ab protein in genetically modified maize

An innovative covalent microsphere immunoassay, based on the usage of fluorescent beads coupled to a specific antibody, was developed for the quantification of the endotoxin Cry1Ab present in MON810 and Bt11 genetically modified (GM) maize lines. In particular, a specific protocol was developed to assess the presence of Cry1Ab in a very broad range of GM maize concentrations, from 0.1 to 100% [weight of genetically modified organism (GMO)/weight]. Test linearity was achieved in the range of values from 0.1 to 3%, whereas fluorescence signal increased following a nonlinear model, reaching a plateau at 25%. The limits of detection and quantification were equal to 0.018 and 0.054%, respectively. The present study describes the first application of quantitative high-throughput immunoassays in GMO analysis.     

Denitrification with and without maize plants (Zea mays L.) under irrigated field conditions

The study was conducted under irrigated field conditions to examine the effect of maize plants on denitrification. Both planted and unplanted field plots received 150kgNha^–1 as urea. In a third treatment, which was also planted and received urea at 150kgNha^–1, the soil nitrate N content was brought up to equal to that in the unplanted plots by applying additional doses of N as calcium nitrate. Soil cores were collected 24 and 72h after irrigation and the denitrification rate was measured by the acetylene inhibition method. Nitrate-N content, aerobically mineralizable C, microbial biomass carrying capacity and denitrification potential were also studied on field-moist soil. Maize plants grown under field conditions always had the potential to increase denitrification in conditions of both high and low water-filled porosity. When nitrate-N content of the planted soil decreased due to plant uptake, denitrification was reduced in the planted soils. However, when nitrate-N uptake by plants was compensated through additional doses of nitrate fertilizer, denitrification was always higher in planted than unplanted soil. The stimulatory effect of plants on denitrification was observed at both high and low soil nitrate-N concentrations, though it was more pronounced at high nitrate-N levels. The effect of plants on denitrification and related parameters was confined to the root zone. Received: 15 April 1996     

Brassinolide effects on maize (Zea mays L.) growth and yield under waterlogged conditions

Our aim was to assess if Brassinolide (BR) could ameliorate stress caused by waterlogging on maize. Two BR levels (with and without), two maize varieties [Ikom White (IKW) and Obatanpa-98 (Oba-98)] and three growth stages [control (WL_o), seedling stage (WL₁), and tasseling stage (WL₂)] were studied under waterlogging lasting 10 days. Maize growth and development were significantly (p?≤?.05) reduced by waterlogging stress under WL₁ than WL₂. Waterlogging stress at WL₁ adversely affected (p?≤?.05) the protein and relative water contents. The nitrogen (N) content among the plant partitions (leaves, stems, and grains) were reduced (p?≤?.05) at both silking and harvest. The beneficial effect of BR was more pronounced in Oba-98 with higher protein contents, dry matter yield, N-uptake and harvest index than IKW. Oba-98 was also better yielding than IKW. Thus, in a waterlogged soil, treatment of maize plants with BR at WL₁ could induce some tolerance.     

Compositional equivalency of Cry1F corn event TC6275 and conventional corn (Zea mays L.)

Maize (Zea mays L.) plants have been transformed to express a Cry1F insecticidal crystal protein originally isolated from Bacillus thuringiensis Berliner. This protein controls lepidopteran pests of maize, including the European corn borer, Ostrinia nubilalis (Hübner). As part of the safety assessment for crops containing transgenes, a compositional analysis of the food and feed is conducted. This analysis is designed to detect unintended changes in the nutrient and antinutrient content of the raw commodities produced by the crop due to the insertion of the genes into the genomic DNA of the plant (pleotropic effects). Samples of transgenic and nontransgenic maize forage and grain were collected from six field sites located in the U.S. and Canada. Forage samples were analyzed for proximates and minerals, and grain was further analyzed for fatty acids, amino acids, vitamins, secondary metabolites, and antinutrients. Results demonstrated that maize expressing the Cry1F protein was equivalent to nontransgenic maize with respect to these important components. Comparison of the variability within the nontransgenic and transgenic hybrid, as compared to composition values reported in the literature, suggest that factors other than transgenes may contribute more substantially to the composition of crops.     

N-utilization by maize (Zea mays L.) as influenced by crop rotation and field pea (Pisum sativum L.) residue management

Abstract. A five year field experiment was conducted to assess the influence of crop rotation and field pea residue incorporation into the soil on maize yield. The data indicated a 30% increase in maize yield grown in rotation with field pea compared to when it was grown after wheat and a further increase of 35% when field pea residues were incorporated into the soil. The effect of field pea and residue incorporation was greater in the presence of fertilizer nitrogen indicating the enhanced capacity of the crop to utilize N from the residue. Legume residue management in sub-tropical regions of the world, having coarse textured soils low in organic matter, could help to increase the yield of cereals besides saving some of the expensive fertilizer input.     

Diversification of maize (Zea mays L.) through teosinte (Zea mays subsp. parviglumis Iltis & Doebley) allelic

Genetic Resources and Crop Evolution - Zea mays ssp. parviglumis is the progenitor of maize and assume to have tolerance against various biotic and abiotic stresses. It...     

Variation among maize (Zea mays L.) accessions of Bendel State,Nigeria

Summary Variability in maize zein protein band mobilities in sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) was characterized to classify 27 maize accessions (OTUs) collected from Bendel State, Nigeria. The classification of the OTUs was achieved using two numerical procedures: average linkage cluster analysis and principal component analysis (PCA). Five clusters were delineated by the cluster analysis whereas the PCA complemented the cluster analysis by separating the OTUs with yellow kernels into one group and OTUs with early maturity into another. OTUs from the same geographical contiguity commonly grouped together. However some regional overlappings of the OTUs occurred. Results of the PCA revealed that zein bands that stained less intensely more strongly separated the OTUs into various clusters than did those that stained more intensely.     

Nutrient concentration and yield of maize (Zea mays L.) after vetch (Vicia sativa L.) in conventional and reduced tillage systems

The effects of tillage on plant availability nutrients are critical to develop nutrient management strategies to optimize yield. The objective of this study was to evaluate differences in the concentration of 19-nutrient and yield after 4-year of conventional [moldboard plough (MT)] and two reduced [rototiller (ST) and chisel (CT)] tillage systems in maize (Zea mays L.) after winter vetch (Vicia sativa L.) in double crop one year. Three tillage systems were compared from 2005 to 2008 in area located in the western region of Turkey under semi-arid Mediterranean clay-loam soils. Nutrients were determined in maize leaves, stems, roots and soils. Results show that the concentration of macronutrients were found to be higher in leaves and stems of maize than roots in three tillage systems of all years, while the opposite was true for micronutrients. Among the macro and micro-nutrients, there was no effect of tillage on nutrient concentration in all maize tissues for sulfur, magnesium, sodium and copper (S, Mg, Na and Cu). However, the nitrogen, potassium, calcium, boron, zinc, manganese, iron, aluminum, barium, cadmium, cobalt, chromium, nickel, lead and selenium (N, K, Ca, B, Zn, Mn, Fe, Al, Ba, Cd, Co, Cr, Ni, Pb and Se) were affected by tillage. ST increased N in stems of 2006 compared with other systems. K in roots of 2006 was 52 and 30% greater in CT than in ST and MT, respectively, while ST and MT of 2007 resulted in 38 and 41% greater than CT. In two of four years, ST contributed to higher grain yield compared with other systems. In general, ST can effectively contribute to increase maize yield following winter vetch compared with MT under this region. Results suggested the need for different management systems associated with reduced tillage including rotation, particularly for basic nutrients. Further, results showed similarities and differences with other studies under tillage with maize following winter vetch.     

Influence of rhizosphere bacteria on morphological characteristics of maize seedlings (Zea mays L.)

The effect of a rhizosphere microflora on some morphological and physiological plant characteristics was studied with maize seedlings grown for five days in a mineral nutrient solution. In the presence of the microorganisms the root dry weight is lower than that of axenically grown plants due to a smaller diameter of the primary root. In addition, the root content of some vitamins and sugars is affected. Pure cultures of rhizosphere bacteria were isolated and their influence on morphological characteristics of the maize plant was classified. Whereas one culture retards the overall plant development, the remaining nine cultures exert a significant influence only on specific morphological parameters. These results are discussed as an indication of the participation of phytohormones in interactions between roots and bacteria.     

Cell wall composition in juvenile and adult leaves of maize (Zea mays L.)

Many leaf characteristics vary with position along the culm in maize (Zea mays L.) due to the existence of vegetative phase change and heteroblasty. The objective of this work was to determine if differences in cell wall composition exist among developmental phases and between Cg1, a developmental mutant, and wild-type maize. In one experiment, the middle third of fully elongated leaf blades from lower and upper regions of the shoot was harvested (midribs removed) and analyzed for several cell wall components. Averaged over five inbreds (De811, Ia5125, Mo17, P39, and Wh8584), lower leaf blades had higher levels of xylose and lower levels of total uronosyls, glucose, arabinose, and galactose (P < 0.05) than did upper leaf blades. With the exception of glucose, upper and lower leaves of Cg1 plants varied in the same manner as their near-isogenic siblings, except cell walls of Cg1 plants were more "juvenile" than cell walls of wild-type siblings at the same leaf stage. These data support the hypothesis that Cg1 delays but does not eliminate the transition from juvenile-vegetative to adult-vegetative phase. In a second experiment, juvenile (leaves 3 and 5), transition (leaf 7), and adult (leaves 9 and 11) leaves from inbreds B73 and De811 were harvested and analyzed as in the first experiment. As leaf number rose, total cell wall content of sample dry matter, total neutral sugars, glucose, xylose, and ester-linked monomers of p-coumaric acid and total ferulates including ferulate dimers increased linearly while total uronosyls acids, arabinose, and galactose declined linearly (P < 0.05). Glucose and xylose are major cell wall components released from cellulose and xylans after acid hydrolysis. Pectin, a minor component of grass cell walls, is composed of galacturonosyls, arabinose, and galactose. Secondary cell wall deposition increased between leaves 3 and 11 in a heteroblastic series, due to either increased cell wall content concomitant with decreased cell lumen size, changes in proportion of cell types (i.e., sclerenchyma), or a combination of these factors.     

不同种植密度下丛枝菌根真菌对玉米磷吸收的影响

【目的】利用土著丛枝菌根真菌(arbuscular mycorrhizal fungi,AM真菌)与作物形成互惠互利的共生关系提高作物对土壤磷的利用效率是解决农业生产中磷供需矛盾的主要途径之一,本研究在大田玉米不同种植密度条件下,研究AM真菌对玉米根系的侵染及磷吸收作用,为揭示集约化玉米高效获取磷的机理提供理论依据。【方法】以大田作物玉米的两种种植密度(5104 plants/hm2和9104 plants/hm2)体系为研究对象,在田间原位埋设PVC管装置,通过测定菌丝生长室中的菌丝密度和有效磷耗竭来确定不同种植密度体系条件下AM真菌对玉米磷吸收的作用。【结果】相对于低密度种植群体,高密度群体显著降低了玉米拔节期土壤有效磷的耗竭量,同时增加了玉米地上部的磷含量,即磷吸收效率,增幅达20%; 在玉米拔节期,增加种植密度使根际的根外菌丝生物量(菌丝密度)降低了4%,而非根际土壤中的根外菌丝生物量(菌丝密度)增加了37%; 高密度玉米种植密度群体中AM真菌的根外菌丝对土壤有效磷耗竭的贡献增加了22%。【结论】集约化玉米生产中土著AM真菌依然帮助植株从土壤中吸收有效磷; 高密度体系下玉米对磷的吸收更加依赖于AM真菌。高密度种植增加AM真菌对玉米的侵染、 根外菌丝量和对土壤有效磷的吸收。     

A comparison of the availability of three zinc sources to maize (Zea mays L.) under greenhouse conditions

 The objective of this study was to compare the availability of Zn from granular ZnSO₄, a granular Zn humate-lignosulfonate (ZnHL), and liquid Zn ethylenediaminetetraacetate (ZnEDTA). The two granular sources were applied directly (“as is”) and also powdered and mixed with the soil. A calcareous sandy loam soil with low available Zn was used. Zn rates were 0, 4, 8, 16, and 32 mg pot^–1, and two crops of maize (Zea mays L.) were grown. For the first crop, all Zn sources provided an excellent dry matter response when powdered and mixed with the soil. The ZnEDTA was superior with regard to Zn uptake. The availability of ZnSO₄ granules was almost zero for the first crop. The availability of the ZnHL complex was better than ZnSO₄ when applied in granular form, but not when the two materials were powdered and mixed with the entire soil mass, suggesting that availability differences between these sources were due to physical, not chemical, factors. After mixing with the soil, only small differences existed between the Zn sources for the second crop. All sources provided for a good dry matter and Zn uptake response, and all Zn sources were about the same in increasing diethylenetriaminepentaacetate (DTPA)-extractable Zn levels at the end of the experiment. This research suggests that the three Zn materials were equal long-term Zn sources, but that the short-term response varied dramatically, depending on how the materials were applied. Received: 8 March 1999     

Kinetics of 15N-labelled nitrate uptake by maize (Zea mays L.) root segments

Abstract

Isotherms and kinetic constants of nitrate uptake by excised root segments from the apical root zone of 6-d-old maize seedlings pretreated with nitrate were investigated using ¹⁵N-labelled nitrate. The isotherms were resolved into two systems namely a multiphasic saturable system at substrate concentrations lower than 2 mol m^?-3 and a linear system at higher concentrations. The detailed analysis of the multiphasic saturable system suggested the existence of at least three phases, which followed the Michaelis-Menten kinetics. The I _max and K _m of each phase increase from the lower phase to the upper phase. The distance from the root tip and the presence of stele affected considerably the linear system but only slightly the saturable system.     

玉米幼胚离体培养和植株再生

系统研究了不同材料、不同培养基对玉米幼胚愈伤组织诱导及继代分化、植传再生的影响，结果发现，8个玉米自交系幼胚均能诱导出愈伤组织。但在继代过程中差异较大。将能继代的愈伤组织转入分化培养基进行生根培养。5010和综3自交系获得了再生植株。并且植株移栽可成活。