Threshold effects on the lectin-mediated aggregation of synthetic glycolipid-containing liposomes

Cholesterol analogs containing sugar residues linked by spacer groups to the cholesterol O can be incorporated into egg yolk lecithin small unilamellar liposomes. The synthetic glycolipid analogs distribute evenly on both sides of the bilayer. These liposomes are aggregated by the appropriate lectin. For example, when the sugar residue is a β-galactoside the liposomes are aggregated by ricin and when it is an α-mannoside they are aggregated by Con A. The lectin-mediated aggregation of these liposomes is reversed by the addition of the appropriate sugar. The rates but not the extents of aggregation of these liposomes are highly sensitive to the amount of glycolipid incorporated. Below approximately 5% glycolipid incorporation the rate of the lectin-mediated aggregation of these liposomes is exceedingly slow, whereas above this level rapid aggregation proceeds. At all concentrations studied the synthetic glycolipids are incorporated in a unimodal fashion so that the observed threshold effects cannot be based on possible differences in the manner in which the glycolipids are incorporated at different concentrations. This conclusion is based on (1) studies with galactose oxidase that show that the percentage of galactose oxidation in a liposome prepared from a galactosyl-containing glycolipid is independent of glycolipid concentration, and (2) studies on the aggregation of liposomes containing mixed glycolipids in which the glycolipids are shown to behave independently. The importance of a critical density of membrane-bound receptors in order for aggregation to occur is discussed.     

Toxin production by a rhizobacterialPseudomonas sp. that inhibits wheat root growth

We studied the production of a toxin inhibitory to both winter wheat (Triticum aestivum L.) root growth andEscherichia coli that was produced by a rhizobacterial pseudomonad. Of several carbon sources tested, the most rapid growth and highest toxin concentrations were obtained with glucose, glycerol, or trehalose. Toxin production was repressed with L-cysteine as the nitrogen source. Toxin was produced during the late exponential and early stationary phase of growth by the bacterium and, contrary to studies with other toxins, was unaffected by Fe and P concentrations in the growth medium. Toxin production by the bacterium was the same at growth temperatures of 25 and 15°C while it produced less at 5°C. If the bacterium was able to grow, it produced toxin. No compound tested induced an increase in toxin production indicating toxin production is constitutive.Contribution from the Agric. Res. Serv., U.S. Dept. of Agriculture in cooperation with the College of Agric. and Home Econ., Res. Ctr., Washington State University, Pullman, Washington 99164.     

Changes in wheat bulb fly (Delia coarctata) populations in East Anglia in relation to crop rotations, climatic data and damage forecasting

Records of wheat bulb fly egg (Delia coarctata) population densities in fields sampled throughout East Anglia in the autumns of 1953–1990 are presented. In descending order of risk, fallow, potato, pea (mainly vining), sugar beet and oilseed rape are the main crops preceding wheat or barley which attract oviposition. A declining trend of egg populations observed over the study period may be associated with climatic changes as well as with the elimination of fallow in the rotation and the dramatic increase in the use of insecticides against the pest. Negative correlations (P < 0.05) in mean annual egg numbers were found with departures from average of July temperature and January air or soil temperature; positive correlations (P<0.05) with departure from average of August raindays. Stepwise regression analysis was used to identify the most important relationships of meteorological variables with mean annual egg numbers, or the proportion of fields sampled with egg numbers in excess of the action threshold of 2.5 million eggs/ha. Up to 59% of the variation in the annual proportion of fields above threshold was accounted for in a regression equation using departures from average of July temperature, August raindays and the percentage of average of October (preceding year) rainfall. Estimated mean annual egg populations and the proportion of fields above threshold showed a good fit with the observed values. The findings are discussed and compared with previous work. The forecasting model may be readily incorporated as a regional risk-prediction component of a knowledge-based system for the management of wheat bulb fly control. Regional forecasts of wheat bulb fly oviposition from this work should be tested and modified as necessary according to experience or changing climatic or agricultural factors.     

Leaching losses of nitrogen from a clay soil under grass and cereal crops in Finland

Summary A 16-plot experimental field was established in 1975 on a clay soil in Jokioinen, Finland. The water discharge through tile drains was measured and its ammonium and nitrate N contents determined for each plot separately. The surface runoff was also measured and analysed. The annual runoff and the N leached from the surface of moderately fertilized (100 kg/ha/y N) cereal plots varied during 1976–1982 from 21 to 301 mm and from 2 to 7 kg/ha, respectively. The discharge of water and leaching of N through subdrains varied from 65 to 225 mm and from 1 to 38 kg/ha, respectively. The highest leaching was probably caused by a previous fallow. The annual N uptake by the crop varied between 41 and 122 kg/ha.Of the fertilizer-N used for cereals, 20% of that applied in the autumn was lost, but only 1 to 4 per cent was lost when applied in the spring. There was much less N leaching from ley than from barley plots, although the former was given twice as much N. The rate of N fertilization had only a very slight effect on N leaching from both ley and barley plots.The results were compared with those obtained in lysimeters filled with clay, silt, sand and peat soils. No definite conclusions can be drawn because the lysimeter experimental data are only for the first year.     

Headgroup dynamics of an integral membrane glycoprotein.

Glycophorin, an integral membrane glycoprotein known to be a receptor for several lectins, has been spin labelled specifically on headgroup terminal sugars. The labelled derivative has been studied in solution and also in various model membranes in an attempt to determine the factors which control headgroup dynamics. Under conditions which mimic those in a living cell the oligosaccharide chains show a uniform, relatively high freedom of motion, with individual sugar correlation times on the order of 6 × 10^?10 sec to 8 × 10^?10 sec depending upon the extent of glycoprotein headgroup involvement with other glycocalyx components. They exhibit no detectable occupancy of lipid or protein hydrophobic domains. Oligosaccharide dynamics are insensitive to factors which act upon that portion of the polypeptide backbone which inserts into the membrane, however a specific recognition event markedly reduces terminal sugar mobility.     

Efficiency in cloning and sequencing using the single-stranded bacteriophage M13

A number of approaches to sequence DNA by the chain termination method are based on cloning into M13 phage vectors and the use of a universal primer. In this paper we investigate some of the factors which influence the speed and efficiency of these approaches. A modification of the template preparation, sequencing reaction, and gel system was used to obtain more reliable and clearer ladder gels. Redundancy and deficiency of shotgun DNA sequencing were reduced by a mapping technique and the use of synthetic primers. The mapping and cloning technique was used to organize the data entry so that the computer time necessary to reconstruct the sequence out of overlaps was reduced.     

Level ofFusarium infection in wheat spikelets related to location and number of inoculated spores

The flowering time is the most susceptible period for primary infection of wheat heads byFusarium spp. During this period spores can be deposited into the opened wheat florets where they may later cause infections. We quantitatively explored the relationship between variables related to the flowering process and the infection level byFusarium graminearum in single spikelets. We imitated open (chasmogamous) and closed (cleistogamous) flowering by injecting well-defined amounts of spores into and between wheat florets. Applying the spores between the florets resulted in weaker disease symptoms and significantly lower amounts ofFusarium mycotoxins. With larger numbers of spores, the disease symptoms became more pronounced and the mycotoxin amounts per spikelet increased significantly. Our results indicate that the probability of primary infection is approximately proportional to the number of spores reaching the open florets during the flowering process. The breeding of wheat lines which flower partially or completely cleistogamously might reduce theFusarium susceptibility in wheat.