A thin-layer chromatographic method for the determination of trace metals in cereal.
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Automation of the clean-up procedure for trichothecenes on a charcoal-alumina column is described. Standard high-performance liquid chromatographic equipment was used for the clean-up step. An acetonitrile-water (84 + 16, v/v) extract of the sample was cleaned up on a column packed with charcoal-alumina-Celite, which was washed with acetonitrile between each sample. The eluates were collected directly in reaction vials and evaporated to dryness. The residual water was removed azeotropically with benzene. The sample was derivatized with 1-(trimethylsilyl)imidazole and analysed by capillary gas chromatography with electron-capture detection.
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Unfermented and fermented maize dough weaning foods prepared by mothers in a Ghanaian village were examined for gram-negative bacilli (GNB) immediately after preparation and during storage to assess the antimicrobial effect of fermentation. GNB were cultured from all samples of unfermented dough (51) and from 16 of 51 samples of fermented dough. The extent of contamination was significantly higher in the unfermented dough than in fermented dough (5.9 [SEM 0.1] vs 4.0 [0.4] log10 colony forming units/g). After 6 h and 12 h storage, a higher proportion of samples of porridge made from unfermented dough contained GNB than did those made with fermented dough (45/51 vs 22/55; 49/51 vs 20/51, respectively) and levels of GNB were significantly higher in the porridge made from unfermented dough after 6 h (4.2 [0.2] vs 3.8 [0.2]). Fermentation of maize dough is an effective method to reduce contamination of maize dough weaning foods with GNB.
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Analysis of the rye cultivar Ailés of several descents derived from crosses between plants carrying specific genotypes and/or chromosome constitutions resulted in the detection of high chromosome (2.05 x 10(-2)) and gene (9.3 x 10(-3)) mutation frequencies. The existence of a transposon system responsible for this instability is suggested.
The nucleotide and deduced amino-acid sequences of a cDNA clone encoding the barley seed protein CMd are described. The sequence is homologous with those of a family of inhibitors of alpha-amylase and trypsin, except for two short insertions. The longest of these (14 residues) is at the junction between the three proposed ancestral regions that comprise this family of proteins, and has limited identity with alpha-amylases of bacterial origin.
Two different cDNA clones (SCL12-1 and SCL12-2) encoding precursors of a chloroplast ribosomal protein with homology to L12 from Escherichia coli were isolated from rye leaf cDNA libraries and sequenced. The corresponding polypeptide of rye chloroplast ribosomes was identified. The sequences for the mature proteins of M(r) 13,447 and 13,609 share 85% amino acid identity. The mature polypeptide of clone SCL12-1 has an amino acid identity of 71%, 72% or 44%, respectively, relative to L12 proteins from spinach, tobacco, or E. coli. Codon usage of the rye L12 cDNAs shows a high preference (97% and 82%) for G or C in the third base position.
Crop samples harvested and stored from three long-term agricultural experiments started in the 1840-1850s at Rothamsted Experimental Station (UK) have been analysed recently for Cd. Increased Cd burden in the soils of the experiments, which have had a range of treatments, originates mainly from atmospheric deposition. Soils treated with farmyard manure (FYM) or, in some cases, applications of phosphate fertilisers, have increased Cd levels. Herbage, wheat and barley grain from the three experiments were analysed by neutron activation analysis (NAA) and graphite furnace atomic absorption spectrometry (GFAAS). Samples were bulked for groups of years between 1860 and 1986, from variously treated plots in each experiment (control or 'nil' treatment, P-fertilised, FYM-amended, NPK-fertilised-limed and unlimed). There were marked differences in Cd concentrations between treatments. For example, uptake of Cd into herbage was greater where P fertiliser had been applied than not, and was greater from unlimed than limed soils. Offtake of Cd (mg ha(-1) year(-1)) was affected by large differences in yield and probably also by other factors. These include changes in botanical composition in the permanent grassland experiment; cultivar changes in the wheat and barley experiments; changes in soil organic matter and soil pH of some plots; changes in atmospheric deposition of Cd through time. All of these potentially confounding factors make the interpretation of results complicated. It is concluded, however, that, with one exception, there is little evidence of a long-term increase in crop Cd concentrations at Rothamsted.
Scanning tunnelling microscopy (STM) has been used to study a seed storage protein of wheat known as gamma-gliadin. The protein was deposited onto highly oriented pyrolytic graphite (HOPG) from solutions of trifluoroethanol (TFE) and 1% acetic acid. Samples were dried down and then scanned in air. Transmission electron microscopy (TEM) was also used to visualise the distribution of protein on the substrate. Small-angle X-ray scattering (SAXS) was used to compare the molecular size and shape obtained with those from the STM images.
Triallate residues in barley seedlings and soil samples were determined by gas chromatography with ion-trap detection. Soil was extracted with methanol on a mechanical shaker, and plants were extracted with acetonitrile in a Sorvall homogenizer. After evaporation of the organic solvents, the residue was dissolved in hexane, and plants extracts were cleaned-up on an alumina column. Gas chromatographic analysis was carried out using a BP-1 fused-silica capillary column with helium as carrier gas. To quantitate residues the total-ion chromatogram was obtained and then the selected-ion monitoring chromatograms were displayed at m/z 86 for triallate and at m/z 154 for the internal standard, methyl-(4-amino-2-chloro)-benzoate. The average recovery through the method from barley and soil samples was always higher than 80%. The limit of detection in the selected-ion mode was 0.01 mg/kg. Barley and soil samples treated with triallate were also analysed. A good agreement was observed between results obtained by this method and by gas chromatography with nitrogen-phosphorus detection.
We report the cloning and characterization of an element (BIS 1) in barley. Related sequences were also found in wheat and rye genomes. BIS 1-related sequences may be some of the more frequent of the complex repeats in the barley genome, and their dispersion throughout the barley genome suggests that they are capable of both mobility and amplification. BIS 1 sequences have been used to study the gross structure of the barley genome. These studies indicate that the genome may be formed from larger genomic structures of tens of kilobases which may be repeated. Surprisingly, these studies also show that these large genomic structures also occur in similar proportions and at similar size distribution in the wheat genome.
Two genes encoding family 11 endo-beta-1,4-xylanases (XylA, XylB) from Fusarium graminearum were cloned and expressed in Escherichia coli. The amount of active endoxylanase in the cytoplasmic soluble fraction was considerably improved by varying different expression parameters, including host strain and temperature during induction. Both recombinant endoxylanases showed a temperature optimum around 35 degrees C and neutral pH optima (around pH 7 and 8 for XylB and XylA, respectively). For the first time this allowed one to test endoxylanases of a phytopathogenic organism for inhibition by proteinaceous endoxylanase inhibitors TAXI and XIP. Whereas XylA and XylB were inhibited by TAXI-I, no inhibition activity could be detected upon incubation with XIP-I. The insensitivity of both F. graminearum endoxylanases towards XIP is surprising, since the latter is typically active against endoxylanases produced by (aerobic) fungi. As F. graminearum is an important phytopathogen, these findings have implications for the role of endoxylanase inhibitors in plant defence.
A total of 344 samples comprising of different vegetables, fodder and grain crops were obtained from a long-term experiment under sewage irrigation. The aerobic bacterial plate counts for vegetables, fodder and grain crops ranged between 2 x 10(6) and 3.5 x 10(7), 6 x 10(6) and 3 x 10(8), 2 x 10(5) and 3.8 x 10(10), respectively, while the corresponding Faecal coliform ranged between < 2 and 9 x 10(5), 9 x 10(2) and 2 x 10(5) and < 2, indicating that the pathogenic loads got reduced below permissible level in the produce that was harvested after sun drying in the field itself, whereas the parts coming in direct contact were the most severely contaminated. The health hazards could be markedly lowered with adoption of some of the low cost practices such as repeated washings, exposure of the produce to sunlight and raising the crops on beds. The coliform counts in vegetables were within permissible limits by two washings with water, exposing these to sunlight for about 4 h and removing the two outmost leaves of cabbage. Also, cutting above some height from ground level (0.10 m) in sorghum reduced the pollution load in fodder crops.
Feeding experiments with diets containing Fusarium toxin-contaminated wheat were conducted to clarify the pathogenesis of enzymatic and histopathological effects of Fusarium toxins on porcine liver cells. A total of 36 prepuberal gilts were divided into four groups and fed diets with increasing proportions of Fusarium toxin-contaminated wheat at a total wheat proportion of 40% over a period of 35 days. The concentrations of the indicator toxins deoxynivalenol (DON) and zearalenone (ZON) which were analyzed by HPLC methods were 210/4, 3070/88, 6100/235, and 9570/358 microg/kg in the diets fed to groups I-IV, respectively. The feeding of mycotoxin-contaminated diets did not cause gross pathological findings in the livers of the animals. Liver tissues were subjected to enzymatic, histological, and ultrastructural examinations. The percentages of the stained areas in periodic acid-Schiff (PAS), Berlin-Blue, and Masson Goldner's trichrome stainings were calculated using the AnalySIS 3.4-system. Significant histopathological findings of alterations with varying degrees in glycogen reduction and increase of hemosiderin particles were found in the liver cells of groups II, III and IV. The thickness of interlobular connective tissue septum in liver cells was significantly increased in groups III and IV. Qualitative ultrastructural alterations were observed in hepatocytes of gilts in groups III and IV. Dependent upon the mycotoxin concentration in the diet, the hepatocytes developed a dose-dependent, extensive, smooth endoplasmic reticulum, exhibited loss of ribosomes, and acquired an increased number of fatty and autophagic vacuoles. However, liver damage as measured by prominent elevated transaminase activities in serum was not detected. Together, the histopathological results provide evidence of liver dysfunction in the absence of clinical signs, especially in pigs fed higher concentrations of Fusarium toxin-contaminated wheat.