The dynamic changes of the levels of volatile organic compounds (VOCs) produced by Bacillus subtilis CF-3 and their biocontrol effects on common fungal pathogens were researched in this study. The results showed that the VOCs in 24-h fermentation liquid (24hFL) of B. subtilis CF-3 inhibited mycelial growth of Botrytis cinerea, Colletotrichum gloeosporioides, Penicillium expansum, Monilinia fructicola, and Alternaria alternata, with a mean inhibition rate of 59.97%. The inhibitory effect on M. fructicola and C. gloeosporioides was the highest; they were therefore selected as target fungal pathogens for further experiments. Based on headspace solid-phase microextraction combined with gas chromatography-mass spectrometry (HS-SPME-GC-MS), 74 potential VOCs were identified during the fermentation: 15 alcohols, 18 ketones, 4 pyrazines, 4 esters, 10 acids, 5 phenols, 3 hydrocarbons, 3 amines, 2 aldehydes, 5 ethers, and 5 other components. At different fermentation times, the type and content of VOCs were different. Most of the potential VOCs (62 VOCs) were identified in the 48hFL. The inhibition rates of all VOCs reached their peaks (73.46% on M. fructicola and 63.63% on C. gloeosporioides) in the 24hFL. Among the identified VOCs, 2,4-di-tert-butylphenol, 1-octanol, and benzothiazole showed significant positive correlations with the rates of M. fructicola and C. gloeosporioides inhibition. Benzoic acid and benzaldehyde showed a significant positive correlation with the rates of M. fructicola inhibition, and anisole and 3-methylbutanal showed a significant positive correlation with the rates of C. gloeosporioides inhibition. In vitro, 2,4-di-tert-butylphenol showed a strong inhibitory effect on both M. fructicola and C. gloeosporioides. In vivo, benzothiazole showed the strongest inhibitory effect on the mycelial extensions of both M. fructicola and C. gloeosporioides, which also led to an increased rate of healthy fruit. The results of the present study clarified that 2,4-di-tert-butylthiophenol and benzothiazole are key inhibitory VOCs produced by B. subtilis CF-3.

In this study, we investigated the effects of volatile organic compounds (VOCs) produced by Bacillus subtilis CF-3 on the growth and development of Colletotrichum gloeosporioides and evaluated the elicitation of active defense responses in harvested litchi fruits. In vitro experiments were conducted to explore the bacteriostatic effect of VOCs in inhibiting pathogenic fungi by means of plate enthalpy test, scanning electron microscopy, transmission electron microscopy, and gas chromatography–mass spectrometry (GC–MS). The results showed that 2,4-di-tert-butylphenol and CF-3 24-h fermentation broth (24hFB) can significantly inhibit the germination of fungal spores, disrupt hyphal and cell morphology, and decrease cell membrane fluidity and integrity, resulting in the changes of indexes. In addition, the bacteriostasis of VOCs in the defensive ability of litchi fruits to C. gloeosporioides was studied, and it was shown that 2,4-di-tert-butylphenol and CF-3 24hFB can inhibit the activity of the pathogenic enzymes (pectinase and cellulase) secreted by C. gloeosporioides to reduce the decomposition of plant tissues, activate antioxidant enzymes (peroxidase, polyphenol oxidase, catalase, and superoxide dismutase) in the fruit to eliminate excessive reactive oxygen species in fruits in order to reduce plant cell damage and activate disease resistance enzymes (phenylalanineammonialyase, chitinases, β-1,3-glucanase) to enhance the resistance of litchi fruits to C. gloeosporioides and inhibit its growth. This study investigated the bacteriostasis of VOCs in inhibiting C. gloeosporioides and inducing the resistance of litchi fruits, providing a theoretical basis for future applications.

In this study, we evaluated the effects of volatile organic compounds (VOCs) produced by Bacillus subtilis CF-3 in inhibiting Monilinia fructicola in vitro and in vivo. In the in vitro experiments, the effect of VOCs on the growth of the pathogenic fungi was explored by using plate enthalpy test; mycelial morphology was studied by scanning electron and transmission electron microscopy; and fatty acid contents in the cell membrane were assessed by gas chromatography-mass spectrometry (GC-MS). The results indicated that treatment with benzothiazole and CF-3 for 24 h, in the form of a fermentation broth (24hFB), significantly inhibited the germination of fungal spores, modified hyphal and cell morphology, and decreased the cell membrane fluidity and integrity. In the in vivo experiments, the effect of VOCs on the defense mechanism of peach fruit toward M. fructicola was studied, and we found that benzothiazole and CF-3 24hFB inhibited the activity of the pathogenic enzymes (pectinase, cellulase) secreted by M. fructicola to reduce the decomposition of plant tissues, activate the antioxidant enzymes (peroxidase, polyphenol oxidase, catalase, and superoxide dismutase) in the fruit to eliminate excessive reactive oxygen species in order to reduce plant cell damage, and trigger the disease-resistant enzymes (phenylalanine ammonia-lyase, chitinases, and β-1,3-glucanase) to enhance the resistance of peach fruit to M. fructicola and inhibit its growth. This study suggests that CF-3 VOCs could activate disease-resistant enzymes to prevent the invasion of pathogenic fungi and induce resistance in peach, thereby providing a theoretical basis for future applications.

Chemical components are one of the most significant traits and attributes of plant tissues, and lead to their different functions. In this study, the composition of Amomun tsao-ko essential oils (AEOs) from different regions was first determined by a combination of gas chromatography–mass spectrometry (GC-MS) and gas chromatography–ion mobility spectrometry (GC-IMS). In total, 141 compounds were identified, of which terpenes and aldehydes were the main groups. Orthogonal partial least square discriminant analysis (OPLS-DA) distinguished the samples from different regions clearly, and the main differences were terpenes, aldehydes, and esters. Meanwhile, AEOs showed strong antibacterial activity against Staphylococcus aureus (S. aureus), and the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) reached 0.20 mg/mL and 0.39–0.78 mg/mL, respectively. From correlation analysis, 1,8-cineole, (E)-dec-2-enal, citral, $\alpha$-pinene, and $\alpha$-terpineol were determined to be the potential antibacterial compounds. This study provides the basis for the variety optimization of A. tsao-ko and its application as a natural food preservative.