Changes in plant metabolism induced by dioxygenase inhibitors and their effect on the epiphytic microbial community and fire blight (Erwinia amylovora) control

Fire blight, caused by the gram negative bacterium Erwinia amylovora, is one of the most destructive bacterial diseases of Pomaceous plants. Therefore, the development of reliable methods to control this disease is desperately needed. This research investigated the possibility to interfere, by altering plant metabolism, on the interactions occurring between Erwinia amylovora, the host plant and the epiphytic microbial community in order to obtain a more effective control of fire blight. Prohexadione-calcium and trinexapac-ethyl, two dioxygenase inhibitors, were chosen as a chemical tool to influence plant metabolism. These compounds inhibit the 2-oxoglutarate-dependent dioxygenases and, therefore, they greatly influence plant metabolism. Moreover, dioxygenase inhibitors were found to enhance plant resistance to a wide range of pathogens. In particular, dioxygenase inhibitors application seems a promising method to control fire blight. From cited literature, it is assumed that these compounds increase plant defence mainly by a transient alteration of flavonoids metabolism. We tried to demonstrate, that the reduction of susceptibility to disease could be partially due to an indirect influence on the microbial community established on plant surface. The possibility to influence the interactions occurring in the epiphytic microbial community is particularly interesting, in fact, the relationships among different bacterial populations on plant surface is a key factor for a more effective biological control of plant diseases. Furthermore, we evaluated the possibility to combine the application of dioxygenase inhibitors with biological control in order to develop an integrate strategy for control of fire blight. The first step for this study was the isolation of a pathogenic strain of E. amylovora. In addition, we isolated different epiphytic bacteria, which respond to general requirements for biological control agents. Successively, the effect of dioxygenase inhibitors treatment on microbial community was investigated on different plant organs (stigmas, nectaries and leaves). An increase in epiphytic microbial population was found. Further experiments were performed with aim to explain this effect. In particular, changes in sugar content of nectar were observed. These changes, decreasing the osmotic potential of nectar, might allow a more consistent growth of epiphytic bacteria on blossoms. On leaves were found similar differences as well. As far as the interactions between E. amylovora and host plant, they were deeply investigated by advanced microscopical analysis. The influence of dioxygenase inhibitors and SAR inducers application on the infection process and migration of pathogen inside different plant tissues was studied. These microscopical techniques, combined with the use of gpf-labelled E. amylovora, allowed the development of a bioassay method for resistance inducers efficacy screening. The final part of the work demonstrated that the reduction of disease susceptibility observed in plants treated with prohexadione-calcium is mainly due to the accumulation of a novel phytoalexins: luteoforol. This 3-deoxyflavonoid was proven to have a strong antimicrobial activity.

Bioremediation of polychlorinated biphenyls (PCBs) polluted marine sediments: a study of the effectiveness of bioaugmentation and biostimulation via microbial electrochemical technologies and polyhydroxyalkanoates

Polychlorinated biphenyls (PCBs) are chemicals largely employed in the industry, banned at the end of the last century yet still persistent in the environment. Bioremediation, namely exploiting bacteria to reduce PCBs’ toxicity, is receiving attention as a promising approach to remediate polluted site in situ. Natural bioremediation is constrained by several factors as the low amount of the required growth substrates (e.g. electron donors, oxygen) and the scarcity of bacteria able to metabolize PCBs. In this regard, use of biodegradable polymers or applied potentials have been demonstrated effective in priming bioremediation of freshwater environments (e.g. river sediments) polluted by chlorinated solvents or PCBs. Yet, little is known regarding the application in marine sediments, where the abundance of anaerobic competitors (i.e. sulfate reducing bacteria) and the different sediment’s features might affect the bioremediation. In this study, polyhydroxyalkanoates (PHAs) and Microbial Electrochemical Technologies (METs) were applied for the first time to prime bioremediation of PCBs polluted marine sediments. The influence of PHAs was studied on the main anaerobic metabolisms and on the microbial community of the heavily polluted sediments coming from the Pialassa della Baiona, a micro-tidal coastal lagoon in Ravenna, and from Mar Piccolo, the marine basin aside Taranto. The impact of METs was deepened by monitoring the physical-chemical parameters and the main anaerobic metabolisms of the sediments coming from Ravenna. The effectiveness of biostimulating with PHAs depended on the features of the treated site, possibly due to the availability of the amendments and to the competition of the indigenous microbial communities. The bioelectrochemical stimulation inhibited the bioremediation process. In both cases, the presence of an inoculated bacterial community was required to perform bioremediation. The collected results led to a comprehensive analysis of the available literature, questioning what could be the further approaches for an effective in situ bioremediation.