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The anthropogenic contribution to the global cadmium flux exceeds natural sources by a factor of three. The most important pathway is the atmosphere; therefore, high cadmium concentrations can be found even in remote areas. On a local level, the increase in cadmium consumption can be observed in increasing concentrations in the soil, plants, and food. The question arises as to what extent the soil-plant-man-waste-soil cycle can be loaded with cadmium in order to function without negative impact on the environment. In Switzerland, 120 tons (t) of cadmium are consumed per year. Of this amount, 25 t end up in municipal solid waste, 3 t in wastewater, and 19 t in precipitation and dry fallout. As a consequence of today's waste management practice (75% incineration, 20% sanitary landfill, 5% composting; 75% of all sewage is purified), the annual input to the soil is 40 t: 18 t concentrated in landfills, 19 t dissipated via the atmosphere, and 3 t directly spread via sewage sludge, compost, and fertilizer on agricultural land. If even distribution were possible, the cadmium content of the soil would theoretically double in 150 years. The accumulation in the soil will increase the cadmium content of plants grown on such a soil. According to a simple model, the level of 3 ppm cadmium in soils should not be surpassed. At such concentrations, plants are likely to contain greater than 0.4 mg Cd/kg, a concentration which can cause toxic effects in long-term experiments. The safe level in food might be even lower. In reality, cadmium is not evenly distributed over Switzerland. According to today's practice, it must be assumed that in only 14 years the use of compost will have enriched soils to such an extent that its cadmium content will prohibit the production of food for human consumption. For sewage sludge, this timespan is 130 years. If heavy metal limits in food are to be observed, the input of such metals to the soil has to be limited. In a steady state, the cadmium input to the soil should be equal to its output via plants, leachate, and erosion. This implies that today's dissipative use of cadmium must be stopped.

A method is described for the direct quantitation of phosphatidylcholine molecular species by reverse phase high-performance liquid chromatography employing flame ionization detection. The method is shown to be applicable to plan phosphatidylcholine. The molecular species are separated with a C18 column eluted in an isocratic mode. Detection by a commercially available flame ionization detector overcomes the problems of detecting underivatized naturally occurring lipids using ultraviolet detectors, and allows direct and rapid mass determination of the resolved molecular species. Detection limits for quantitation are defined.

Spatial and temporal patterns in the fluoride content of native vegetation around two aluminium smelters in the Hunter Valley were studied between 1982 and 1985. Foliage samples were collected every month from dominant tree, shrub and herb species located up to 15 km from each smelter and, after washing, were analysed for their fluoride concentrations. At Kurri Kurri, an established smelter, fluoride emissions varied between 10 and 15 tonnes per month, or 1.8 and 2 kg [corrected] per tonne Al produced. At Tomago, they increased rapidly during start-up, peaked at 15.2 tonnes per month (0.7 kg per tonne Al), and then fell to around 10 tonnes per month. Fluoride isopleths for Angophora bakeri and a histogram for A. costata, revealed that the main areas of impact were to the immediate north and north-east of Kurri Kurri and immediate north and south-east of Tomago. Although foliar fluoride concentrations greater than background levels extended 3 km from Kurri Kurri and 1 km from Tomago, fluoride-induced, visible injury was more limited in extent. Close to the smelters tree species accumulated more foliar fluoride than shrub species, which in turn accumulated more foliar fluoride than herb species. Foliar fluoride concentrations in tree and herb species were lowest in summer; spring peaks were also apparent at Tomago.

A glucan of high molecular weight isolated from stale sugar-cane, and previously shown to have a marked effect on sucrose crystallisation processes-1, is a relatively linear dextran. Approximately 96-97% of its D-glucose residues are involved in (1 yields 6)-alpha-D linkages and constitute the linear backbone of the polymer. The remaining 3-4% of D-glucose residues form branch-points by (1 yields 3)-alpha-D linkages. The periodate-oxidation technique, which has been extensively used by other workers to determine dextran structure, gave erroneous results when applied to the dextran from stale sugar-cane.

Microsomal b-type hemoprotein designated, cytochrome b555 of C.roseus seedlings was solubilized using detergents and purified by a combination of ion exchange chromatography and gel filtration to a specific content of 18.5 nmol per mg of protein. The purified cytochrome b555 was homogeneous and estimated to have an apparent molecular weight of 16500 on SDS-PAGE. The absorption spectrum of the reduced form has major peaks at 424, 525 and 555 nm. The alpha-band of the reduced form is asymmetric with a pronounced shoulder at 559 nm. The spectrum of the pyridine ferrohemochrome shows absorption peaks at 557, 524 and 418 nm indicating that the cytochrome has protoheme prosthetic group. The purified cytochrome is autoxidizable and does not combine with carbon monoxide, azide or cyanide. It is reducible by NADH in the presence of NADH-cytochrome b555 reductase partially purified from C.roseus microsomes.

Two methods for analysis of acid-labile sulfide and zero-valence sulfur in plant extracts containing chlorophyll as well as ionic and/or nonionic detergents are presented. Both methods are based on the conversion of sulfide into methylene blue. In the first method an ethyl acetate extraction step is used to remove chlorophyll and its degradation products which otherwise prevent spectrophotometric quantitation of methylene blue. The second assay method employs 35S-labeled plant extracts. This method, which involves thin-layer chromatography and autoradiography, is potentially more sensitive than the spectrophotometric assay in detecting acid-labile sulfide and zero-valence sulfur.

Employing specific dye-stain followed by fluorescence microscopy, evidence was obtained for the presence of amyloplast nucleoids in protoplasts as well as in the amyloplasts isolated from the suspension- cultured cells of sycamore (Acer psuedoplatanus L.), which are devoid of chlorophylls and grow heterotrophically. We have isolated and partially characterized amyloplast DNA. The presence of six homologous sequences to chloroplast genes encoding (i) apoprotein of P700, (ii) 32 kDa protein (photogene 32), (iii) alpha subunit of CF1, (iv) large subunit of RuBisCO, (v) beta and epsilon subunits of CF1 and (vi) 16S ribosomal RNA, has been revealed by hybridization experiments, using gene probes from maize chloroplast DNA.

A biomonitoring study was made of the atmospheric deposition of arsenic and selenium across northern Canada utilizing Sphagnum fuscum moss. Intensive sampling was carried out adjacent to the smelter at Rouyn-Noranda, Quebec. An average background concentration of arsenic in the moss of 0.66 mg kg-1 was determined, while most selenium concentrations were below the detection limit of 0.2 mg kg-1. Elevated arsenic concentrations were found in the vicinity of the mining and smelting areas of Flin Flon, Manitoba, and Atikokan, Ontario. High concentrations of selenium were detected only near Rouyn-Noranda. Both arsenic and selenium occurred in moss at concentrations lower than found in Canadian soils.

The radioactivity of various terrestrial vegetation leaves characteristic of Mediterranean countries has been measured after the Chernobyl accident. In addition, we paid particular attention to lichens and seaweed which are considered as bioindicators of radioactive contamination. Most measurements were performed non-destructively using both coaxial and planar HPGe detectors. For odd mass radionuclides having low energy lines, such as 125Sb or 141Ce the sensitivity of the planar HPGe detector is better than the coaxial detector. The concentration of long-lived fission nuclides remaining three months after the accident were found to be enhanced in needle form leaves and in lichens. The seaweed Sphaerococcus exhibits a strong specific activity for iodine and ruthenium elements and poor concentration for caesium nuclides. The activity ratios of different isotopes of the same element measured in vegetation samples agree well with values found in airborne aerosols by other authors. The activation nuclide 110mAg is found in all samples with the same ratio 110mAg/137Cs = (1.0 +/- 0.2).10(-2) as in the soil deposition.

A procedure which uses hot methanolic HCl to digest fresh tissue and simultaneously convert the fatty acids of the leaf lipids to the corresponding methyl esters is described. Extraction of the fatty acid methyl esters into a small volume (0.3 ml) of hexane means that a sample for GLC analysis can be taken directly from the tube used for the digestion/methylation reaction. The procedure provides a fatty acid analysis which is comparable to that obtained by a more conventional technique involving separate extraction, saponification, and methylation steps, but the overall yield is reduced by 10-20%. The analysis can be made quantitative by including an internal standard with the tissue sample.

Acetohydroxyacid synthase (AHAS), also known as acetolactate synthase, has received attention recently because of the finding that it is the site of action of several new herbicides. The most commonly used assay for detecting the enzyme is spectrophotometric involving an indirect detection of the product acetolactate. The assay involves the conversion of the end product acetolactate to acetoin and the detection of acetoin via the formation of a creatine and naphthol complex. There is considerable variability in the literature as to the details of this assay. We have investigated a number of factors involved in detecting AHAS in crude ammonium sulfate precipitates using this spectrophotometric method. Substrate and cofactor saturation levels, pH optimum, and temperature optimum have been determined. We have also optimized a number of factors involved in the generation and the detection of acetoin from acetolactate. The results of these experiments can serve as a reference for new investigators in the study of AHAS.

Concentration of lead have been determined in three species of small mammal, Sorex araneus, Clethrionomys glareolus and Apodemus sylvaticus at roadside and control sites. The results are considered in relation to the lead content of the diet and the air, and it is concluded that the diet provides far the major pathway for lead exposure, even at the roadside site. Bone lead concentrations are found to be consistent with estimated lead exposure and metabolism.

A method is described for the liquid scintillation counting of 14C in plant tissues. Samples are fixed, in the scintillation vial, in a solution of ethanol and acetic acid (3:1) and decolorized with commercial bleach before the addition of scintillation liquid. The method was compared to other techniques of tissue oxidation or digestion and found to be equally effective at least with thin tissue samples. The technique is simple, rapid, and inexpensive and does not result in loss of 14C.

Problems encountered in obtaining reliable analytical data by HPLC for the free nucleotide constituents of plant tissues are considered and methods of overcoming them experimentally assessed. Major problems include suppression of residual phosphatase activity during extraction, and removal of pigments, phenolics, alkaloids, and other uv-absorbing nonnucleotides, prior to HPLC. An optimal combination of extraction and pre-HPLC purification techniques is discussed which, in combination with HPLC by anion exchange, yields quantitatively reliable data. The optimized procedure involves extraction with a monophasic mixture of methanol: chloroform:formic acid:water and purification of the nucleotide extract by a batch treatment with poly-N-vinylpyrrolidone, followed by ligand-exchange chromatography. The main HPLC separation uses mu Bondapak NH2 in a linear phosphate gradient and gives good resolution of all the commonly occurring plant nucleotides in a single chromatographic run.

An enzyme-linked immunosorbent assay (ELISA) which used 4-methylumbelliferyl phosphate as an enzyme substrate was used to quantify two plant cytokinins. This assay detected as little as 0.03 pmol (approximately 10 pg) of cytokinin in microplate wells coated with a cytokinin-ovalbumin conjugate. The method measured competition between free cytokinin and the bound conjugate for reaction with monoclonal anticytokinin antibodies and used a standard curve prepared by use of known amounts of free cytokinin to quantify hormone levels in unknown samples. Standard curves which consisted of logit/log plots of fluorescence units versus picomoles of competing cytokinin measured from 0.03 to 256 pmol (approximately 10-85,000 picograms) of zeatin riboside (ZR) or isopentenyl adenosine. The fluorescence ELISA was compared with radioimmunoassay for the quantification of ZR in wheat (Triticum aestivum L., cultivar Stephens) seed samples. This fluorescence ELISA method is recommended for use in combination with a fractionation method, such as HPLC, to quantify cytokinins present in plant extracts.

After sampling over regular intervals from 1985 to 1987 the heavy metals Cd, Cu, Pb and Zn were quantitatively determined by atomic emission spectrometry/inductively coupled plasma in above-ground parts of the moss Polytrichum formosum (Hedw.). All metals present a typical summer-winter oscillation (seasonal variation), which is much greater than the "inter-stand" variation of Polytrichum formosum within the forest ecosystem under investigation. To obtain comparable results on a regional or global scale it is suggested to collect all samples of Polytrichum formosum for biomonitoring purposes in the last week of September.

Two giant pandas were used to assess the utilization of bamboo as a feedstuff. Three 1-week-long digestion trials were conducted during which feed intake and fecal output were recorded. Passage of digesta was measured from both fluid and particulate markers administered at feeding. Results indicate that the giant panda, although highly specialized for the consumption of bamboo, is a very inefficient digester of bamboo. The pandas consumed up to 6% of body weight in dry matter per day, with bamboo dry matter digestibility averaging less than 20%. Apparent digestion coefficients for the structural carbohydrates of bamboo (27% for hemicellulose and 8% for cellulose) indicate that, unlike most herbivores, pandas do not rely heavily on the microbial degradation of plant material to fulfill their nutritional requirements. Additionally, the passage of digesta through the gastrointestinal tract of the panda was extremely rapid, with complete clearance of markers in less than 12 hours. The giant panda seems to have specialized to a feed source high in plant fiber without extensive modification of the digestive tract by selectivity in feeding, effective mastication, ingestion of large quantities, digestion of cellular contents rather than cell walls and rapid fecal excretion of the undigested residue.

Triclopyr, after esterification, is shown to be a suitable candidate for detection by gas chromatography-electron-capture negative-ion chemical ionization mass spectrometry forming a characteristic carboxylate anion which offers a high detection sensitivity. A detection limit of 70 fg reaching the ionizer is indicated. Low backgrounds and an absence of chemical interferences are shown for vegetation extracts, using a simple method of extraction and derivatisation. A similar behaviour is demonstrated for 2,4-D and 2,4,5-T.

Three phenolic constituents of Magnolia grandiflora L. were shown to possess significant antimicrobial activity using an agar well diffusion assay. Magnolol, honokiol, and 3.5'-diallyl-2'-hydroxy-4-methoxybiphenyl exhibited significant activity against Gram-positive and acid-fast bacteria and fungi. The minimum inhibitory concentrations were determined for each compound using a twofold serial dilution assay.

A thermospray liquid chromatography-mass spectrometry interface connected to a double-focusing magnetic sector instrument has been applied successfully to the solution of a number of analytical problems in food and agricultural research. Examples of the qualitative analysis of some anti-nutrients, plant growth control hormones, biotechnology products and a plant polyphenol are presented and discussed.

A phytochemical investigation of an alcoholic extract of the petioles of Nelumbo lutea resulted in the identification of the alkaloids N-methylasimilobine, anonaine, and roemerine. The alkaloids nuciferine, armepavine, N-nornuciferine, and N-norarmepavine, previously previously reported in the whole plant, were also identified.

Antisera raised against a highly conserved amino acid sequence (G alpha-common peptide) of animal Gs alpha, Gi alpha, Go alpha and Gt alpha recognize, in plasma membranes of several higher plants, sets of proteins of Mr = 37 and 31 kDa (Vicia faba), 36 and 31 kDa (Arabidopsis thaliana) and 38 and 34 kDa (Commelina communis). The A. thaliana proteins were solubilized and partially purified. They bind [35S]GTP gamma S with high affinity (apparent Kd approximately 10 nM) and, with lower affinity, GTP but not the other nucleotides tested (ATP, CTP, ITP, UTP).

This paper describes a new glycosyl-sequencing method. This method was made possible by the ability to fractionate complex mixtures of peralkylated oligosaccharides by reversed-phase, high-pressure liquid chromatography. The fractionation ability of the reversed-phase system allows the isolation and subsequent unambiguous identification by g.l.c.-m.s. of disaccharides, almost all trisaccharides, and, in some cases, tetrasaccharides generated by successive partial acid hydrolysis, reduction, and ethylation of a permethylated, complex carbohydrate. As these small oligosaccharides overlap within the unhydrolyzed, complex carbohydrate, the oligosaccharide sequences may be pieced together, and, with the glycosyl-linkage composition of the intact complex carbohydrate, can be used to determine the glycosyl sequence of the complex carbohydrate. The details of the sequencing method are illustrated by the elucidation of the glycosyl sequences of three complex carbohydrates. These examples demonstrate the wide variety of complex carbohydrates whose structures can be ascertained by the new sequencing technique. Two of the examples are the commercially available polysaccharides, lichenan and xanthan, whose structures have already been reported. The other example is a nonasaccharide derived from xyloglucan, a structural polymer of plant cell-walls. The glycosyl residues of the complex carbohydrates studied include hexosyl, deoxyhexosyl, pentosyl, glycosyluronic, and pyruvic acetal-substituted hexosyl residues. It will be demonstrated that the new glycosyl-sequencing technique is not compromised by the presence, in the carbohydrate to be analyzed, of glycosyl linkages possessing very different acid labilities. Two major advantages of this sequencing technique are that it is relatively rapid and that it requires only milligram quantities of carbohydrate.

A simplified and sensitive method for the analysis of eight carbamate insecticide residues in foliage, forest soil and fish tissue by direct gas-liquid chromatography (GLC) is described. After fortification, the carbamate residues were extracted from the natural substrates by homogenization in ethyl acetate. The interfering co-extratives present in the crude extracts were removed by filtration through Whatman GF/A glass microfibre after coagulation. The carbamate residues were re-extracted into dichloromethane and directly analyzed intact by GLC with a Tracor Model 702 nitrogen-phosphorus detector. The rate of recovery for aminocarb, carbofuran, carbaryl, methomyl, mexacarbate, pirimicarb and propoxur was better than 80% at 5.0-ppm and 0.50-ppm levels. Slightly less than 60% was recovered for methiocarb.

A simple soluble protein extraction method was developed for glanded cotton (Gossypium hirsutum L.) tissues. Gossypol, a major component of glands, is known to crosslink and precipitate proteins in cotton tissue homogenates. Established phenolic removal reagents were evaluated as gossypol binding agents and found to be less than effective in enhancing cotton leaf-soluble protein recovery. Several other amines, including a number of affinity support bound amines, were tested and found relatively ineffectual when compared with urea as cotton protein protectants. Urea and (NH4)2SO4, the next most active agent found in the study, were compared on both whole-leaf homogenates and artificial mixtures containing known quantities of poly-L-lysine and a clathrate of gossypol and acetic acid. Urea treatment resulted in both an increased number of stained bands and more concentrated staining of bands than any other treatment on polyacrylamide gels. Selected enzymes demonstrated increased activities in urea homogenates compared with other treatments.