Fertile transgenic barley by particle bombardment of immature embryos

Transgenic, fertile barley (Hordeum vulgare L.) from the Finnish elite cultivar Kymppi was obtained by particle bombardment of immature embryos. Immature embryos were bombarded to the embryonic axis side and grown to plants without selection. Neomycin phosphotransferase II (NPTII) activity was screened in small plantlets. One out of a total of 227 plants expressed the transferred nptII gene. This plant has until now produced 98 fertile spikes (T₀), and four of the 90 T₀ spikes analyzed to date contained the nptII gene. These shoots were further analyzed and they expressed the transferred gene. From green grains, embryos were isolated and grown to plantlets (T₁). The four transgenic shoots of Toivo (the T₀ plant) produced 25 plantlets as T₁ progeny. Altogether fifteen of these T₁ plants carried the transferred nptII gene as detected with the PCR technique, fourteen of which expressed the nptII gene. The integration and inheritance of the transferred nptII gene was confirmed by Southern blot hybridization. Although present as several copies, the transferred gene was inherited as a single Mendelian locus into the T₂ progeny.     

Biomass allocation and nutrient use in fast-growing woody and herbaceous perennials used for phytoremediation

This study assessed the suitability of two deciduous woody perennials (Salix spp. and Populus spp.) and two summer green herbaceous perennials (Phragmites australis and Urtica dioica) for purification of nutrient enriched wastewater. The main hypothesis tested was that species with a particular trait combination of high relative growth rate (RGR), low nutrient productivity (A) and high mean residence time (MRT) of nutrients would be most effective in accumulating nutrients. The nitrogen and phosphorus use efficiency at the whole plant level was analysed. Four treatments comprising two possible phytoremediation substrates (municipal wastewater and landfill leachate) and two control plant nutrition situations (balanced nutrient solution and pure water) were applied in four replications to the four plant species. Generally, all four species studied showed a high RGR and a low P productivity in the balanced nutrient solution treatment, while the opposite (low RGR and high P productivity) was seen in the phytoremediation substrate and pure water treatments. The general conclusion is that if P is present in marginal proportions in the wastewater, a vegetation filter with Phragmites would have an advantage since biomass and nutrient accumulation in Phragmites does not decrease as much during phytoremediation as that in deciduous woody perennials.     

Clonal growth in a species-rich grassland: Results of a 20-year fertilization experiment

We investigated the influence of fertilization on the abundance of species with different clonal growth characteristics using the data from a 20-year fertilization experiment from the Laelatu wooded meadow, in Estonia. The experiment comprised four different fertilization treatments and created a gradient of nitrogen availability. The vegetation composition was recorded every year by measuring the proportions of aboveground biomass for all species. For each species, four parameters of vegetative propagation were measured: speed of ramet vegetative mobility (annual increment of rhizome length), frequency of rhizome branching, placement of branches, and ramet life span. The weighted average of each parameter was calculated for each plot both at the beginning and at the end of the experiment using the relative abundances of the species in the plot as weights. The community changes resulting from the fertilization are reflected in the significant changes of the average values of all studied clonal growth parameters. Increased levels of phosphorus and potassium led to a community with an increased average vegetative mobility and rhizome branching. Both of these traits, however, declined with the increasing availability of nitrogen. The proportion of species with long-living ramets in the community decreased with the increase in the productivity irrespective of the fertilizer used. There was a strong positive correlation between the average ramet life span of the community and the number of species on the plot. We concluded that fertilization increased the ramet turnover rate in this meadow community and reduced species richness. Thus, our results contradict the prediction of a higher ramet turnover rate in species-rich compared to the species-poor grasslands.     

Association between cagA and vacA genotypes and pathogenesis in a Helicobacter pylori infected population from South-eastern Sweden

ABSTRACT: BACKGROUND: Chronic gastritis, peptic ulcer disease, and gastric cancer have been shown to be related toinfection with Helicobacter pylori (H. pylori). Two major virulence factors of H. pylori,CagA and VacA, have been associated with these sequelae of the infection. In this study, totalDNA was isolated from gastric biopsy specimens to assess the cagA and vacA genotypes. RESULTS: Variations in H. pylori cagA EPIYA motifs and the mosaic structure of vacA s/m/i/dayregions were analysed in 155 H. pylori-positive gastric biopsies from 71 individuals usingPCR and sequencing. Analysis of a possible association between cagA and vacA genotypesand gastroduodenal pathogenesis was made by logistic regression analysis. We found that H. pylori strains with variation in the number of cagA EPIYA motif variants present in the samebiopsy correlated with peptic ulcer, while occurrence of two or more EPIYA-C motifs wasassociated with atrophy in the gastric mucosa. No statistically significant relation betweenvacA genotypes and gastroduodenal pathogenesis was observed. CONCLUSIONS: The results of this study indicate that cagA genotypes may be important determinants in thedevelopment of gastroduodenal sequelae of H. pylori infection. In contrast to other studies,vacA genotypes were not related to disease progression or outcome. In order to fullyunderstand the relations between cagA, vacA and gastroduodenal pathogenesis, themechanisms by which CagA and VacA act and interact need to be further investigated.     

Catalytic convergence of manganese and iron lipoxygenases by replacement of a single amino Acid

Lipoxygenases (LOXs) contain a hydrophobic substrate channel with the conserved Gly/Ala determinant of regio- and stereospecificity and a conserved Leu residue near the catalytic non-heme iron. Our goal was to study the importance of this region (Gly(332), Leu(336), and Phe(337)) of a lipoxygenase with catalytic manganese (13R-MnLOX). Recombinant 13R-MnLOX oxidizes 18:2n-6 and 18:3n-3 to 13R-, 11(S or R)-, and 9S-hydroperoxy metabolites (～80-85, 15-20, and 2-3%, respectively) by suprafacial hydrogen abstraction and oxygenation. Replacement of Phe(337) with Ile changed the stereochemistry of the 13-hydroperoxy metabolites of 18:2n-6 and 18:3n-3 (from ～100% R to 69-74% S) with little effect on regiospecificity. The abstraction of the pro-S hydrogen of 18:2n-6 was retained, suggesting antarafacial hydrogen abstraction and oxygenation. Replacement of Leu(336) with smaller hydrophobic residues (Val, Ala, and Gly) shifted the oxygenation from C-13 toward C-9 with formation of 9S- and 9R-hydroperoxy metabolites of 18:2n-6 and 18:3n-3. Replacement of Gly(332) and Leu(336) with larger hydrophobic residues (G332A and L336F) selectively augmented dehydration of 13R-hydroperoxyoctadeca-9Z,11E,15Z-trienoic acid and increased the oxidation at C-13 of 18:1n-6. We conclude that hydrophobic replacements of Leu(336) can modify the hydroperoxide configurations at C-9 with little effect on the R configuration at C-13 of the 18:2n-6 and 18:3n-3 metabolites. Replacement of Phe(337) with Ile changed the stereospecific oxidation of 18:2n-6 and 18:3n-3 with formation of 13S-hydroperoxides by hydrogen abstraction and oxygenation in analogy with soybean LOX-1.