Chemical and spectroscopic analyses ((13)C cross-polarization-magic angle spinning NMR and attenuated total reflection Fourier transform infrared spectroscopies) were carried out on the wood of Vitis vinifera cv. Sangiovese with brown-red discoloration and black streaks caused by esca disease. The analyses of the brown-red wood revealed the destruction of hemicelluloses and noncrystalline cellulose as well as modifications in the pectic and ligninic wood fractions. The pectic fraction consisted of carbohydrates associated with polyphenols. The lignin fraction exhibited only a few changes in the aromatic systems and a partial demethylation, and it appeared to be associated with condensed phenolic components probably arising from response polyphenols. The degradation of hemicelluloses and noncrystalline cellulose in brown-red wood, where the pathogens Phaeoacremonium aleophilum and Phaeomoniella chlamydospora prevail with respect to the other fungus Fomitiporia mediterranea, was consistent with reports on the degradative activity of such fungi in vitro carried out on model substrates. The observed alterations could also be attributed to the radical oxidation process caused by the oxidative response of defense itself triggered by infection, as suggested by the accumulation of postinfectional compounds. The analyses of wood tissue with black streaks showed less marked deterioration; here, an increase in pectic and phenolic substances, which probably accumulate in the xylem vessels as a response to the infection, was observed.
The influence of different combinations of illumination and shaking on the growth dynamics, pathogenicity and toxin production of the fungus Fusarium oxysporum f. sp. orthoceras, a biocontrol agent of Orobanche cumana, was studied. The fastest biomass accumulation was obtained under shaking, with or without illumination, with the highest biomass obtained after 3-4 weeks of growth. The biological activity of chloroform extracts of the culture filtrate was characterised: it contained at least two main toxic metabolites that caused necrosis and wilting of various plants and led to mortality of germinating seeds of O. cernua, O. aegyptiaca, O. ramosa and O. cumana. The highest toxic activity of the chloroform extract was obtained under illumination without shaking after 3-4 weeks of growth. The two toxic metabolites were purified and identified as fusaric acid (FA) and 9,10-dehydrofusaric acid (DFA). Both FA and DFA production began in the first week of growth, increasing gradually to their maxima after 4 weeks. The highest level of pathogenic activity of the fungus was obtained after three or more weeks of fungal growth. It can be concluded that in order to produce high levels of toxin and pathogenic activity, the fungus should be grown under illumination without shaking for 4 weeks
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