As an edible and medicinal fungus, Dictyophora indusiata is well-known for its morphological elegance, distinctive taste, high nutritional value, and therapeutic properties. In this study, eighteen compounds (1–18) were isolated and identified from the ethanolic extract of D. indusiata; four (1–4) were previously undescribed. Their molecular structures and absolute configurations were determined via a comprehensive analysis of spectroscopic data (1D/2D NMR, HRESIMS, ECD, and XRD). Seven isolated compounds were examined for their anti-inflammatory activities using an in vitro model of lipopolysaccharide (LPS)-simulated BV-2 microglial cells. Compound 3 displayed the strongest inhibitory effect on tumor necrosis factor-α (TNF-α) expression, with an IC50 value of 11.9 μM. Compound 16 exhibited the highest inhibitory activity on interleukin-6 (IL-6) production, with an IC50 value of 13.53 μM. Compound 17 showed the most potent anti-inflammatory capacity by inhibiting the LPS-induced generation of nitric oxide (NO) (IC50: 10.86 μM) and interleukin-1β (IL-1β) (IC50: 23.9 μM) and by significantly suppressing induced nitric oxide synthase (iNOS) and phosphorylated nuclear factor-kappa B inhibitor-α (p-IκB-α) expression at concentrations of 5 μM and 20 μM, respectively (p < 0.01). The modes of interactions between the isolated compounds and the target inflammation-related proteins were investigated in a preliminary molecular docking study. These results provided insight into the chemodiversity and potential anti-inflammatory activities of metabolites with small molecular weights in the mushroom D. indusiata.
Neem (Azadirachta indica) extract is well-known as a natural pesticide for the control of agricultural pests. Azadirachtin A and its structural analogues are considered as active compounds. However, the amounts of azadirachtins varies in neem extracts, providing a variety of insecticidal activities. In this study, a novel method of automated online solid-phase extraction coupled with liquid chromatography/quadrupole-time-of-flight mass spectrometry (SPE-LC–Q-TOF–MS) was developed and validated for simultaneous quantification of five azadirachtins (azadirachtins A, B, D, H and I) in seed and leaf extracts of A. indica. Different experimental parameters (such as SPE cartridge, injection volume and washing step) were optimized. The optimized SPE-LC–Q-TOF–MS method showed good recovery (82.0–102.8%), linearity (r2 ≥ 0.9991) and precision (0.83–4.83%). The limit of detections (LODs) for the five analytes ranged from 0.34 to 0.76 ng mL−1. The validated method was successfully applied for determination of the analytes in the neem leaves and seeds from different locations and a neem formulation. The online SPE-LC–Q-TOF–MS method was found to be a simple, precise and accurate and can be used as a powerful tool for quality control of neem extracts or its formulations.
To clarify the residue formation of prothioconazole and its main metabolite in wheat plants, the uptake, translocation, and metabolism of prothioconazole in wheat roots and leaves were investigated by nutrient solution culture and the spraying method. The results showed that prothioconazole and its metabolites showed a trend of increasing and then decreasing in wheat plants under two treatment methods, and the concentration of prothioconazole and its metabolites was higher in the high-concentration group (5000 μg/mL) than in the low-concentration group (1000 μg/mL). The transferability from stem to leaf was stronger than that from root to stem. In the nutrient solution culture, prothioconazole and its metabolites were mainly enriched in wheat roots. The concentration of prothioconazole in wheat roots increases with the increase in prothioconazole concentration and was significantly higher than the prothioconazole concentration on stems and leaves. In wheat leaves in the spraying method, prothioconazole and its metabolites were conducted from leaves to stems and roots up to the nutrient solution. Prothioconazole-desthio was detectable in wheat nutrient solution, while prothioconazole was not detected. Analysis of actual samples of 9 wheat grains and 28 flours showed that the residues of prothioconazole and its metabolites met the maximum residue limit (0.1 mg/mL) set in China and by the Codex Alimentarius Commission. The results will provide a theoretical basis for the scientific use of prothioconazole and food security assurance.
Combined seed treatment with neonicotinoids and fungicides offers a potential control measure for pest management at the wheat seeding stage. In this study, a novel, highly-efficient seed-coating agent was prepared using thifluzamide, fludioxonil, and clothianidin as its active components and other additives (abbreviated to TFC). Laboratory experiments and field trials revealed a positive effect on germination, plant height, and root length, with 90% control efficiency on wheat sharp eyespots and aphid infestations. Meanwhile, the distribution of thifluzamide, fludioxonil, and clothianidin residues in the wheat plants at harvest was below 0.05 mg/kg both at the recommended dosage and at 2.0 times the recommended dose. Furthermore, an artificial soil assay of biotoxicity in earthworms revealed a low level of toxicity at LC50 > 10 mg/kg. Overall, these findings suggest that TFC has the potential to control major soil-borne diseases and pest infestations in wheat, offering an environmentally-friendly alternative to more toxic pesticides.