Identification of genes involved in fungal responses to strigolactones using mutants from fungal pathogens.

Strigolactones (SLs) as components of root exudates induce hyphal branching of arbuscular mycorrhizal (AM) fungi which is thought to favor the establishment of the beneficial symbiosis. Little is known on how AM fungi respond to SLs. Since AM fungi are poor model systems due to their obligate biotrophism and the lack of genetic transformation protocols, we took advantage of the sensitivity of several phytopathogenic fungi to GR24, a synthetic SLs analog. With the aim to identify the molecular determinants involved in SLs response in AM fungi and assuming conserved mechanisms in the fungal kingdom, we exploited the fungal pathogens Botrytis cinerea and Cryphonectria parasitica, for which mutant collections are available. Exposure of B. cinerea and C. parasitica to GR24 embedded in solid medium led to reduction of fungal radial growth. We set up the screening of a set of well-characterized gene deletion mutants to isolate genotypes with altered responses to SLs. Two B. cinerea mutants (defective of BcTrr1, a thioredoxin reductase and BcLTF1, a GATA transcription factor) turned out to be less responsive to GR24. One feature shared by the two mutants is the overproduction of reactive oxygen species (ROS). Indeed, an oxidizing effect was observed in a B. cinerea strain expressing a redox-sensitive GFP2 in the mitochondrial intermembrane space upon exposure to GR24. ROS and mitochondria are, therefore, emerging as mediators of SLs actions.

Study of suitability of Fricke-gel-layer dosimeters for in-air measurements to characterise epithermal/thermal neutron beams for NCT.

The reliability of Fricke gel dosimeters in form of layers for measurements aimed at the characterization of epithermal neutron beams has been studied. By means of dosimeters of different isotopic composition (standard, containing (10)B or prepared with heavy water) placed against the collimator exit, the spatial distribution of gamma and fast neutron doses and of thermal neutron fluence are attained. In order to investigate the accuracy of the results obtained with in-air measurements, suitable MC simulations have been developed and experimental measurements have been performed utilizing Fricke gel dosimeters, thermoluminescence detectors and activation foils. The studies were related to the epithermal beam designed for BNCT irradiations at the research reactor LVR-15 (Rez). The results of calculation and measurements have revealed good consistency of gamma dose and fast neutron 2D distributions obtained with gel dosimeters in form of layers. In contrast, noticeable modification of thermal neutron fluence is caused by the neutron moderation produced by the dosimeter material. Fricke gel dosimeters in thin cylinders, with diameter not greater than 3mm, have proved to give good results for thermal neutron profiling. For greater accuracy of all results, a better knowledge of the dependence of gel dosimeter sensitivity on radiation LET is needed.

Measurements of 2D distributions of absorbed dose in protontherapy with Gafchromic EBT3 films.

A study of the response of EBT3 films to protons has been carried out with the aim of finding a simple modality to achieve dose images in which the effect of the film sensitivity dependence on radiation LET is amended. Light transmittance images (around 630 nm) were acquired by means of a CCD camera and the difference of optical density was assumed as dosimeter response. The calibration of EBT3 film was performed by means of protons of 173.61 MeV. Some EBT3 films were exposed, in a solid-water phantom, to proton beams of three different energies (89.17 MeV, 110.96 MeV and 130.57 MeV) and the obtained depth-dose profiles were compared with the calculated profiles. From the ratios of calculated and measured Bragg peaks, a trend of the decrease in EBT3 sensitivity with increasing peak depth has been deduced. A method for correcting the data measured with EBT3 films, utilizing the file of irradiation planning data, has been proposed and tested. The results confirm that the method can be advantageously applied for obtaining spatial distribution of the absorbed dose in proton therapy.