Antifungal activity of Ligusticum chuanxiong essential oil and its active composition butylidenephthalide against Sclerotium rolfsii.

BACKGROUND: Peanut stem rot caused by Sclerotium rolfsii is an epidemic disastrous soil-borne disease. Recently, natural products tend to be safe alternative antifungal agents to combat pathogens. RESULTS: This work determined the preliminary antifungal activity of 29 essential oils against S. rolfsii and found that Ligusticum chuanxiong essential oil (LCEO) showed the best antifungal activity, with an EC50 value of 81.79 mg L-1 . Sixteen components (98.78%) were identified in LCEO by gas chromatography-mass spectrometry analysis, the majority by volume comprising five phthalides (93.14%). Among these five phthalides, butylidenephthalide was the most effective compound against S. rolfsii. Butylidenephthalide not only exhibited favorable in vitro antifungal activity against the mycelial growth, sclerotia production and germination of S. rolfsi, but also presented efficient in vivo efficacy in the control of peanut stem rot. Seven days after application in the glasshouse, the protective and curative efficacy of butylidenephthalide at 300 mg L-1 (52.02%, 44.88%) and LCEO at 1000 mg L-1 (49.60%, 44.29%) against S. rolfsii were similar to that of the reference fungicide polyoxin at 300 mg L-1 (54.61%, 48.28%). Butylidenephthalide also significantly decreased the oxalic acid and polygalacturonase content of S. rolfsii, suggesting a decreased infection ability on plants. Results of biochemical actions indicated that butylidenephthalide did not have any effect on the cell membrane integrity and permeability but significantly decreased nutrient contents, disrupted the mitochondrial membrane, inhibited energy metabolism and induced reactive oxygen species (ROS) accumulation of S. rolfsii. CONCLUSION: Our results could provide an important reference for understanding the application potential and mechanisms of butylidenephthalide in the control of S. rolfsii. © 2023 Society of Chemical Industry.

iTRAQ proteomic analysis of the inhibitory effect of 1,6-O,O-diacetylbritannilactone on the plant pathogenic oomycete Phytophthora capsici.

Phytophthora capsici is a highly destructive oomycete of vegetables; its management is challenging due to its broad host range, rapid dispersion, resilient spores and severe fungicide resistance. Identifying an effective alternative fungicide is important for the control of P. capsici. 1,6-O,O-diacetylbritannilactone (ABLOO), one of the secondary metabolites of Inula Britannica, showed a favorable inhibitory activity against P. capsici at different developmental stages, with a sensitivity order as follows: sporangia formation (30.45 mg/L) > zoospore discharge (77.69 mg/L) > mycelial growth (93.18 mg/L) > cystospore germination (591.48 mg/L). To investigate the mode of action of ABLOO in P. capsici, iTRAQ-based quantitative proteomic analysis was performed by comparing the expression levels of proteins in the control and ABLOO-treated (400 mg/L, inhibition rate of 80%) mycelial groups. A total of 65 downregulated and 75 upregulated proteins were identified in the proteomic analysis. Functional enrichment analyses showed that proteins with transmembrane transport activity were significantly inhibited, while proteins involved in energy production were significantly increased, including proteins involved in ubiquinone and other terpenoid-quinone biosynthesis, oxidative phosphorylation, and glycolysis/gluconeogenesis. The morphological results indicated that ABLOO treatment could decrease the thickness of the cell walls of P. capsici mycelia. Correspondingly, biochemical results showed that ABLOO treatment reduced the ß-1,3-glucan contents (the key component of the cell wall of P. capsici) and increased the cell membrane permeability of P. capsici. ABLOO may exhibit antioomycete activity by destroying the cell membrane of P. capsici. This study provides new evidence regarding the inhibitory mechanisms of ABLOO against P. capsici.

Comparative Analysis of Botrytis cinerea in Response to the Microbial Secondary Metabolite Benzothiazole Using iTRAQ-Based Quantitative Proteomics.

Benzothiazole is a microbial volatile compound with strong antifungal activity against the phytopathogenic fungus Botrytis cinerea, but its mode of action against fungi remains largely unknown. Understanding the molecular mechanisms underlying its activity could aid the design and synthesis of similar compounds against pathogenic fungi. Based on the results of morphological and antifungal activity assays, B. cinerea was exposed to 2.5 µl/liter of benzothiazole for 12, 24, and 48 h, and an isobaric tags for relative and absolute quantitation-based quantitative proteomic analysis showed that 378 out of 5,110 identified proteins were differentially expressed proteins (DEPs). The majority of these DEPs were associated with carbohydrate metabolism, oxidation reduction processes, and energy production. Further analysis showed that benzothiazole inhibited mitochondrial membrane organization and decreased the mitochondrial membrane potential of B. cinerea. In addition, the key enzymes of the glyoxylate cycle were downregulated after benzothiazole treatment, and a biochemical analysis indicated that inhibition of the glyoxylate cycle by benzothiazole blocked nutrient availability and interfered with adenosine triphosphate generation. This study provides markers for future research of the molecular responses of B. cinerea to benzothiazole stress.

Proteomic profile of the Bradysia odoriphaga in response to the microbial secondary metabolite benzothiazole.

Benzothiazole, a microbial secondary metabolite, has been demonstrated to possess fumigant activity against Sclerotinia sclerotiorum, Ditylenchus destructor and Bradysia odoriphaga. However, to facilitate the development of novel microbial pesticides, the mode of action of benzothiazole needs to be elucidated. Here, we employed iTRAQ-based quantitative proteomics analysis to investigate the effects of benzothiazole on the proteomic expression of B. odoriphaga. In response to benzothiazole, 92 of 863 identified proteins in B. odoriphaga exhibited altered levels of expression, among which 14 proteins were related to the action mechanism of benzothiazole, 11 proteins were involved in stress responses, and 67 proteins were associated with the adaptation of B. odoriphaga to benzothiazole. Further bioinformatics analysis indicated that the reduction in energy metabolism, inhibition of the detoxification process and interference with DNA and RNA synthesis were potentially associated with the mode of action of benzothiazole. The myosin heavy chain, succinyl-CoA synthetase and Ca+-transporting ATPase proteins may be related to the stress response. Increased expression of proteins involved in carbohydrate metabolism, energy production and conversion pathways was responsible for the adaptive response of B. odoriphaga. The results of this study provide novel insight into the molecular mechanisms of benzothiazole at a large-scale translation level and will facilitate the elucidation of the mechanism of action of benzothiazole.