Procedures allowing the transformation of a range of European elite wheat (Triticum aestivum L.) varieties via particle bombardment.

Ten current European wheat varieties were transformed at efficiencies ranging from 1-17% (mean 4% across varieties) following modifications in particle bombardment and tissue culture procedures. All plants surviving phosphinothricin selection were screened for uidA and bar gene activity, and for the presence of marker gene sequences by PCR analysis. A minimum of 35% plant 'escape' frequency was achieved with selection on 4 mg l(-1) gluphosinate ammonium after shoot initiation. Mean co-transformation frequency with various genes-of-interest was 66%. The estimated number of insertions of the uidA gene in 25 lines were; 1-2 in 32%, 3-5 in 52%, and 6-8 in 16% of lines. In T(1) progenies, marker genes segregated in a Mendelian fashion in 50% of 39 lines analysed, as determined by transgene activity assays. Based on PCR analysis, it appeared that in some lines the occurrence of distorted segregation was due to poor transmission of the transgenes.     

Ochratoxin A displaces claudins from detergent resistant membrane microdomains

Ochratoxin A (OchA) is a food-borne mycotoxin with multiple effects in vivo. Previously, we have demonstrated that the toxin can significantly impair the barrier function of the gut epithelial cell line, Caco-2. Barrier disruption involved loss of claudins 3 and 4, but not claudin 1 from the tight junction complex. In this study, we demonstrate for the first time, that OchA is able to remove claudins 3 and 4 from the detergent insoluble membrane microdomains associated with the tight junctions. However, cholesterol distribution within the microdomain was unaffected by the toxin. In addition, the thiol antioxidant, N-acetyl cysteine, preserved the microdomain localisation of claudins and also the barrier function of Caco-2 cells. This work suggests that OchA-mediated barrier toxicity is due to removal of claudins from detergent insoluble membrane microdomains. Moreover, loss of microdomain association may be due to oxidative events.     

The high-latitude terrestrial carbon sink: a model analysis

A dynamic, global vegetation model, hybrid v4.1 ( Friend et al. 1997 ), was driven by transient climate output from the UK Hadley Centre GCM (HadCM2) with the IS92a scenario of increasing atmospheric CO₂ equivalent, sulphate aerosols and predicted patterns of atmospheric N deposition. Changes in areas of vegetation types and carbon storage in biomass and soils were predicted for areas north of 50°N from 1860 to 2100. Hybrid is a combined biogeochemical, biophysical and biogeographical model of natural, potential ecosystems. The effect of periodic boreal forest fires was assessed by adding a simple stochastic fire model. Hybrid represents plant physiological and soil processes regulating the carbon, water and N cycles and competition between individuals of parameterized generalized plant types. The latter were combined to represent tundra, temperate grassland, temperate/mixed forest and coniferous forest. The model simulated the current areas and estimated carbon stocks in the four vegetation types. It was predicted that land areas above 50°N (about 23% of the vegetated global land area) are currently accumulating about 0.4 PgC y^?1 (about 30% of the estimated global terrestrial sink) and that this sink could grow to 0.8–1.0 PgC y^?1 by the second half of the next century and persist undiminished until 2100. This sink was due mainly to an increase in forest productivity and biomass in response to increasing atmospheric CO₂, temperature and N deposition, and includes an estimate of the effect of boreal forest fire, which was estimated to diminish the sink approximately by the amount of carbon emitted to the atmosphere during fires. Averaged over the region, N deposition contributed about 18% to the sink by the 2080 s. As expected, climate change (temperature, precipitation, solar radiation and saturation pressure deficit) and N deposition without increasing atmospheric CO₂ produced a carbon source. Forest areas expanded both south and north, halving the current tundra area by 2100. This expansion contributed about 30% to the sink by the 2090 s. Tundra areas which were not invaded by forest fluctuated from sink to source. It was concluded that a high latitude carbon sink exists at present and, even assuming little effect of N deposition, no forest expansion and continued boreal forest fires, the sink is likely to persist at its current level for a century.     

Cytochrome P450105D1 (CYP105D1) from Streptomyces griseus: heterologous expression, activity, and activation effects of multiple xenobiotics.

The open reading frame of CYP105D1, a soluble cytochrome P450 from Streptomyces griseus, was cloned behind the tac promoter of the bacterial expression vector pSPg1910L and expressed in Escherichia coli. The recombinant protein retained normal spectral characteristics having a Soret peak at 448 nm in the reduced carbon monoxide difference spectrum. CYP105D1 was active, obtaining reducing equivalents from endogenous E. coli ferredoxin and ferredoxin reductase redox partners present in E. coli. In vitro activity studies revealed CYP105D1 to catalyse the NADH- and NADPH-dependent oxidation of the xenobiotic substrates benzo[a]pyrene, erythromycin, warfarin, and testosterone. Furthermore, this activity could be stimulated in the presence of either alpha-benzoflavone or beta-benzoflavone in an analogous manner to that reported for mammalian P450 forms including human liver cytochrome P4503A4 (CYP3A4). The system produces an alternative to whole-cell biotransformation of xenobiotic for the production of drug metabolites and an experimental system for probing the structural features of a cytochrome P450 with a broad substrate range.     

Climate change and pollutant impacts on Scottish vegetation

Summary

By the 2050s the UK is projected to be about 1.6°C warmer, when the atmospheric CO₂ concentration will be 525 ppmv. These changes will have profound effects on the Scottish flora and fauna. Vegetation primary productivity will increase, except in dry regions, and the productivity of upland forest plantations may increase by several Yield Classes. The spread of plant species may be less than expected, but a number of slow-growing ‘stress-tolerant’ species, including montane/alpine species, are likely to be lost. Nitrogen deposited as a result of emission of NO_X from vehicles and NH₃ from agriculture is now a major source of acidity, and problems of acidification and eutrophication are linked. Despite reductions in sulphur emissions, critical loads of acid deposition are likely to be exceeded for soils in most of the Scottish uplands until at least 2005. Critical levels affecting tree growth may be exceeded where forests are in cloud for 10% of the time in areas of the Great Glen. Much of the Scottish uplands receives 25–30 kg N ha^-1 yr^-1, which may be causing change in species composition. Background tropospheric ozone concentrations are increasing. Much of the Scottish uplands experiences mean summer ozone concentrations exceeding those in southern England, but with fewer exceedances of critical levels. However, many crops and some sensitive native species are probably being adversely affected.     

Studies on pre-incubation conditions in the seeded agar plate well inhibition assay

Pre-incubation at 4°C increases the zone of bacterial growth inhibition produced by methanolic extracts of freshwater algae in the agar diffusion assay. Preloading of wells with methanol increases the inhibition zone produced by water-insoluble antibiotics.     

A computer program for processing data from amino acid analysis and for the calculation of molecular weights from those data.

A computer program is described for the calculation of the complete amino acid composition of a protein from the analytical data. The program also derives a molecular weight on the basis of the amino acid composition. The use of the program for the determination of the molecular weights of the liver carboxylesterases of chicken, horse, ox, and sheep is described.