Due to hydrophobicity, plant essential oil components transcinnamaldehyde and thymol exert broad-spectrum antimicrobial activities by interfering with the structures and functions of microbial cell membranes, yet their antifungal properties against food-spoilage yeast Zygosaccharomyces rouxii are still unclear. This work aimed to elucidate their antifungal activities and mechanisms against Z. rouxii. Minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) were measured using broth dilution method to evaluate the antifungal activities. Membrane permeability, potential and integrity, intracellular ATP content, and intracellular pH (pH in ) were determined, and membrane damage was observed by scanning electron microscope (SEM) to investigate the antifungal mechanisms. The MICs of transcinnamaldehyde and thymol were 0.3125 and 0.0625 mg/ml, respectively, and the MFCs were 1.25 and 0.125 mg/ml, respectively. Increased membrane permeability, membrane hyperpolarization, decreased membrane integrity, reduced intracellular ATP content, and lowered pH in were found after transcinnamaldehyde or thymol treatment at the MIC and MFC levels, and thymol exhibited more severe influence on cell membrane. SEM observation further confirmed that trans-cinnamaldehyde caused cell membrane shrinkage and invagination, while thymol resulted in perforation in cell membrane. These results suggest that trans-cinnamaldehyde and thymol exert anti-Z. rouxii activities by inducing cell membrane damage and subsequent intracellular homeostasis disruption. Cellular contents leakage and intracellular homeostasis disruption probably proceed more rapidly after thymol inducing perforation in the cell surface, thereby thymol exerting better anti-Z. rouxii activity.

This work mainly aimed to evaluate the feasibility of Fourier transform-near infrared spectroscopy (FT-NIRS) combined with chemometrics in early identification of freshness decay of fresh-cut kiwifruit during simulated cold chain storage, with organoleptic evaluation as a reference. By linear discriminant analysis, the freshness decay could be identified after only 2 days of cold storage, corresponding to freshness level of 3.41 ± 0.27 N (hardness), 0.70 ± 0.05 g/kg (total acid), 8.62 ± 0.06 g/100 g (reducing sugars), 62.04 ± 1.03 mg/100 g (vitamin C) and 2.05 ± 0.11 log 10 CFU/g (total plate count).Organoleptic evaluators could not perceive the freshness decay that occurred after 2 days of cold storage. Moreover, the freshness decay could be well quantitatively predicted by partial least squares regression, with low RMSE p (0.18-05.42) and high R 2 (0.90-0.96). FT-NIRS appears to be a promising option for early warning of the freshness decay of fresh-cut kiwifruit during cold chain storage, thereby preventing serious spoilage and ensuring fresh fruits for consumers.

This work aimed for the first time to provide detailed insights into thymol and trans-cinnamaldehyde's mechanisms of action on the food-spoilage yeast Zygosaccharomyces rouxii and offers evidence in favor of the activation of an apoptosis-like phenotype. The action mechanisms of thymol and transcinnamaldehyde were investigated by the measurement of a series of typical apoptotic features using flow cytometer or microplate reader. Moreover, quantitative reverse transcription PCR (QRT-PCR) was performed to investigate the effects of thymol and trans-cinnamaldehyde on the transcription of key regulators of apoptosis in Z. rouxii. The results indicated that the treatment of Z. rouxii with thymol or trans-cinnamaldehyde (minimum inhibitory and subinhibitory concentrations) triggered reactive oxygen species (ROS) accumulation, elevated intracellular Ca 2+ level, and depolarized mitochondrial membrane potential (MMP) coupled with hallmarks of apoptosis including mitochondrial cytochrome c (cyt c) release, metacaspase activation, phosphatidylserine (PS) exposure, and DNA fragmentation. Moreover, thymol or trans-cinnamaldehyde treatment upregulated the transcription of proapoptotic regulators including Yca1, Dnm1, Nuc1, Ndi1, and Mmi1 and downregulated the transcription of antiapoptotic regulators of Fis1 and Cdc48, further confirming the apoptosis induced by thymol and trans-cinnamaldehyde in Z. rouxii. In summary, thymol and transcinnamaldehyde probably induced apoptosis through a metacaspase-dependent mitochondrial pathway in Z. roxuii. These findings provide theoretical support for the development of safe and efficient agents used in food preservation.