The Agrobacterium-mediated transformation of common wheat (Triticum aestivum L.) and triticale (x Triticosecale Wittmack): role of the binary vector system and selection cassettes

The influence of two binary vector systems, pGreen and pCAMBIA, on the Agrobacterium-mediated transformation ability of wheat and triticale was studied. Both vectors carried selection cassettes with bar or nptII driven by different promoters. Two cultivars of wheat, Kontesa and Torka, and one cultivar of triticale, Wanad, were tested. The transformation rates for the wheat cultivars ranged from 0.00 to 3.58% and from 0.00 to 6.79% for triticale. The best values for wheat were 3.58% for Kontesa and 3.14% for Torka, and these were obtained after transformation with the pGreen vector carrying the nptII selection gene under the control of 35S promoter. In the case of the bar selection system, the best transformation rates were, respectively, 1.46 and 1.79%. Such rates were obtained when the 35S::bar cassette was carried by the pCAMBIA vector; they were significantly lower with the pGreen vector. The triticale cultivar Wanad had its highest transformation rate after transformation with nptII driven by 35S in pCAMBIA. The bar selection system for the same triticale cultivar was better when the gene was driven by nos and the selection cassette was carried by pGreen. The integration of the transgenes was confirmed with at least three pairs of specific starters amplifying the fragments of nptII, bar, or gus. The expression of selection genes, measured by reverse transcriptase polymerase chain reaction (RT-PCR) in relation to the actin gene, was low, ranging from 0.00 to 0.63 for nptII and from 0.00 to 0.33 for bar. The highest relative transcript accumulation was observed for nptII driven by 35S and expressed in Kontesa that had been transformed with pGreen

Transgene × Environment Interactions in Genetically Modified Wheat

BACKGROUND: The introduction of transgenes into plants may cause unintended phenotypic effects which could have an impact on the plant itself and the environment. Little is published in the scientific literature about the interrelation of environmental factors and possible unintended effects in genetically modified (GM) plants. METHODS AND FINDINGS: We studied transgenic bread wheat Triticum aestivum lines expressing the wheat Pm3b gene against the fungus powdery mildew Blumeria graminis f.sp. tritici. Four independent offspring pairs, each consisting of a GM line and its corresponding non-GM control line, were grown under different soil nutrient conditions and with and without fungicide treatment in the glasshouse. Furthermore, we performed a field experiment with a similar design to validate our glasshouse results. The transgene increased the resistance to powdery mildew in all environments. However, GM plants reacted sensitive to fungicide spraying in the glasshouse. Without fungicide treatment, in the glasshouse GM lines had increased vegetative biomass and seed number and a twofold yield compared with control lines. In the field these results were reversed. Fertilization generally increased GM/control differences in the glasshouse but not in the field. Two of four GM lines showed up to 56% yield reduction and a 40-fold increase of infection with ergot disease Claviceps purpurea compared with their control lines in the field experiment; one GM line was very similar to its control. CONCLUSIONS: Our results demonstrate that, depending on the insertion event, a particular transgene can have large effects on the entire phenotype of a plant and that these effects can sometimes be reversed when plants are moved from the glasshouse to the field. However, it remains unclear which mechanisms underlie these effects and how they may affect concepts in molecular plant breeding and plant evolutionary ecology

Il bilancio consolidato

Il capitolo si propone di introdurre il lettore alla tematica del bilancio consolidato, illustrandone le finalità conoscitive, gli ambiti di applicazione e le principali metodologie e tecniche di redazione sulla base dei principi contabili internazionali

Fusarium graminearum exploits ethylene signalling to colonize dicotyledonous and monocotyledonous plants

Ethylene signalling affects the resistance of dicotyledonous plant species to diverse pathogens but almost nothing is known about the role of this pathway in monocotyledonous crop species. Fusarium graminearum causes Fusarium head blight (FHB) of cereals, contaminating grain with mycotoxins such as deoxynivalenol (DON). Very little is known about the mechanisms of resistance/susceptibility to this disease. * Genetic and chemical genetic studies were used to examine the influence of ethylene (ET) signalling and perception on infection of dicotyledonous (Arabidopsis) and monocotyledonous (wheat and barley) species by F. graminearum. Arabidopsis mutants with reduced ET signalling or perception were more resistant to F. graminearum than wild-type, while mutants with enhanced ET production were more susceptible. These findings were confirmed by chemical genetic studies of Arabidopsis, wheat and barley. Attenuation of expression of EIN2 in wheat, a gene encoding a core component of ethylene signalling, reduced both disease symptoms and DON contamination of grain. Fusarium graminearum appears to exploit ethylene signalling in both monocotyledonous and dicotyledonous species. This demonstration of translation from model to crop species provides a foundation for improving resistance of cereal crops to FHB through identification of allelic variation for components of the ethylene-signalling pathway