The antifungal effects of thyme, cinnamon bark and clove bud essential oils (EOs) were investigated in vitro on Colletotrichum acutatum mycelial growth, conidial germination, appressoria formation, and in vivo on strawberry fruit disease incidence. All tested EOs, incorporated in potato-dextrose agar, inhibited C. acutatum mycelial growth, and had a fungistatic effect at concentration 667 μl/l of medium. Volatiles of cinnamon bark, thyme and clove bud EOs completely prevented conidial germination at the lowest concentrations of 1.53, 15.3 and 76.5 μl/l of air, respectively, and disabled appressoria formation at concentration of 1.53 μl/l of air. On inoculated strawberry fruit, thyme and cinnamon bark EO volatiles reduced anthracnose incidence at concentrations above 15.3 and 76.5 μl/l of air, respectively. GC-FID and GC-MS analysis showed that major components of thyme EO were p-cymene, thymol, α-terpineol, carvacrol; cinnamon bark EO: transcinnameldehyde, trans-cinnamyl acetate; clove bud EO: eugenol and β-caryophyllene. Our results suggest that volatiles of thyme and cinnamon bark EOs are effective against C. acutatum both in vitro and in vivo.
A polygalacturonase (PG) was extracted and purified from decayed tissue of 'Anjou' pear fruit inoculated with Penicillium expansum. Ammonium sulfate precipitation, gel filtration, and cation exchange chromatography were used to purify the enzyme. Both chromatographic methods revealed a single peak corresponding to PG activity. PG enzyme activity from healthy and wounded pear tissue was undetectable, which supports the claim that the purified PG is of fungal origin. The purified enzyme had a molecular mass of 41 kDa and a pI of 7.8. Activity of the PG was not associated with a glycosylated protein. The enzyme was active over a broad pH range from 3 to 6, with optimal activity at 4.5 in sodium citrate and sodium acetate buffers. The optimal temperature for activity was 37 degrees C but the enzyme was also active at 0, 5, 10, 20, and 50 degrees C. Thin-layer chromatographic analysis of PG hydrolysis products showed that the enzyme exhibits endo- and exo-activity. The purified enzyme macerated tissue in vitro causing approximately 30% reduction in mass of pear plugs compared with approximately 17% reduction for apple. Additionally, it produced 1.5-fold more soluble polyuronides on pear than apple tissue. This work shows for the first time the production of a PG by P. expansum during postharvest decay of pear fruit is different from the previously described PG produced in decayed apple fruit by the same pathogen.
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