The chemical composition of the essential oil obtained from the peel of Bingtang sweet orange (Citrus sinensis Osbeck) was analysed by gas chromatography and gas chromatography ⁄ mass spectrometry (GC ⁄ MS). Twenty-seven components were identified. The monoterpenes and sesquiterpene hydrocarbons with 96.03% (w ⁄ w) of the total oil were the principal compound groups. Among which, limonene was observed dominant (77.49%), followed by myrcene (6.27%), a-farnesene (3.64%), c-terpinene (3.34%), apinene (1.49%), sabinene (1.29%) and other minor components. Results by disc diffusion method and minimum inhibitory concentration (MIC) determination method showed that the essential oil had a wide spectrum of antimicrobial activities against Staphylococcus aureus, Penicillium chrysogenum, Bacillus subtilis, Escherichia coli and Saccharomyces cerevisiae, with their inhibition zones ranging from 14.57 mm to 23.37 mm and the MIC ranging from 4.66 lL mL )1 to 18.75 lL mL )1 .
Citral exhibits strong antifungal activity against Penicillium digitatum. In this study, 41 over-expressed and 84 repressed proteins in P. digitatum after 1.0 µL/mL of citral exposure for 30 min were identified by the iTRAQ technique. The proteins were closely related with oxidative phosphorylation, the TCA cycle and RNA transport. The mitochondrial complex I, complex II, complex III, complex IV and complex V, which are involved in oxidative phosphorylation were drastically affected. Among of them, the activities of mitochondrial complex I and complex IV were apparently suppressed, whereas those of mitochondrial complex II, complex III and complex V were significantly induced. Meanwhile, citral apparently triggered a reduction in the intracellular ATP, the mitochondrial membrane potential (MMP) and glutathione content, in contrast to an increase in the glutathione S-transferase activity and the accumulation of reactive oxygen species (ROS). Addition of exogenous cysteine decreased the antifungal activity. In addition, cysteine maintained the basal ROS level, deferred the decrease of MMP and the membrane damage. These results indicate that citral inhibited the growth of P. digitatum by damaging oxidative phosphorylation and cell membranes through the massive accumulation of ROS.
Ergosterol (ERG) is a potential target for the development of antifungal agents against Penicillium digitatum, the pathogen of green mold in citrus fruits. This study examined the mechanism by which citronellal, a typical terpenoid of Cymbopogon nardus essential oil, acts on ergosterol to exhibit its antifungal activity against P. digitatum. We previously reported that citronellal inhibited the growth of P. digitatum with minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) of 1.36 and 2.72 mg/mL, respectively. In citronellal-treated cells, the membrane integrity and ergosterol contents significantly decreased, whereas lanosterol, which serves as a precursor for ergosterol biosynthesis, massively accumulated. Addition of 150 mg/L of exogenous ergosterol decreased the inhibitory rate of citronellal, restoring the ergosterol content and hence the membrane structure to normal levels, and triggered expression of nearly all ERG genes. Based on our findings, we deduce that citronellal damages the cell membrane integrity of P. digitatum by down-regulating the ERG genes responsible for conversion of lanosterol to ergosterol, the key downregulated gene being ERG3, due to the observed accumulation of ergosta-7,22-dienol.
BACKGROUND: To improve the properties and functionality of bighead carp (Aristichthys nobilis) muscle, bighead carp surimi was inoculated with combinations of Saccharomyces cerevisiae, Lactobacillus and Monascus and fermented, and then bighead carp sausages were prepared. The characteristics of fermented bighead carp surimi and sausages were investigated.
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