Aureobasidium pullulans as biological control agent: modes of action

The postharvest phase has been considered an environment very suitable for successful application of biological control agents (BCAs). However, the tri-interaction between fungal pathogen, host (fruit) and antagonist is influenced by several parameters such as temperature, oxidative stresses, oxygen composition, water activity, etc. that could be determining for the success of biocontrol. Knowledge of the modes of action of BCAs is essential in order to enhance their viability and increase their potentialities in disease control. The thesis focused on the possibility to explain the modes of action of a biological control agent (BCA): Aureobasidium pullulans, in particular the strains L1 and L8, control effective against fruit postharvest fungal pathogen. In particular in this work were studied the different modes of action of BCA, such as: i) the ability to produce volatile organic compounds (VOCs), identified by SPME- gas chromatography-mass spectrometry (GC-MS) and tested by in vitro and in vivo assays against Penicillium spp., Botrytis cinerea, Colletotrichum acutatum; ii) the ability to produce lytic enzymes (exo and endo chitinase and β-1,3-glucanase) tested against Monilinia laxa, causal agent of brown rot of stone fruits. L1 and L8 lytic enzymes were also evaluated through their relative genes by molecular tools; iii) the competition for space and nutrients, such as sugars (sucrose, glucose and fructose) and iron; the latter induced the production of siderophores, molecules with high affinity for iron chelation. A molecular investigation was carried out to better understand the gene regulation strictly correlated to the production of these chelating molucules. The competition for space against M. laxa was verified by electron microscopy techniques; iv) a depth bibliographical analysis on BCAs mechanisms of action and their possible combination with physical and chemical treatments was conducted.

Coupled wellbore-reservoir simulation of thermal effects during multiphase CO2 injection

T2Well-ECO2M is a coupled wellbore reservoir simulator still under development at Lawrence Berkeley National Laboratory (USA) with the ability to deal with a mixture of H2O-CO2-NaCl and includes the simulation of CO2 phase transition and multiphase flow. The code was originally developed for the simulation of CO2 injection into deep saline aquifers and the modelling of enhanced geothermal systems; however, the focus of this research was to modify and test T2Well-ECO2M to simulate CO2 injection into depleted gas reservoirs. To this end, the original code was properly changed in a few parts and a dedicated injection case was developed to study CO2 phase transition inside of a wellbore and the corresponding thermal effects. In the first scenario, the injection case was run applying the fully numerical approach of wellbore to formation heat exchange calculation. Results were analysed in terms of wellbore pressure and temperature vertical profiles, wellhead and bottomhole conditions, and characteristic reservoir displacement fronts. Special attention was given to the thorough analysis of bottomhole temperature as the critical parameter for hydrate formation. Besides the expected direct effect of wellbore temperature changes on reservoir conditions, the simulation results indicated also the effect of CO2 phase change in the near wellbore zone on BH pressure distribution. To test the implemented software changes, in a second scenario, the same injection case was reproduced using the improved semi-analytical time-convolution approach for wellbore to formation heat exchange calculation. The comparison of the two scenarios showed that the simulation of wellbore and reservoir parameters after one year of continuous CO2 injection are in good agreement with the computation time to solve the time-convolution semi-analytical reduced. The new updated T2Well-ECO2M version has shown to be a robust and performing wellbore-reservoir simulator that can be also used to simulate the CO2 injection into depleted gas reservoirs.