A determination method for trace 24-epibrassinolide (EBL) in plant tissues was developed using ultra-high-performance liquid chromatography–tandem mass spectrometry (UPLC-MS/MS). The plant tissue samples were extracted using a methanol–formic acid solution, and the corresponding supernatant was purified with ODS C18 solid-phase extraction column. The extracts were separated using a Zorbax Eclipse Plus C18 (2.1 mm × 50 mm, 1.8 μm) column with methanol and 0.1% formic acid as the mobile phase. The ion source for the mass spectrometry was an electrospray ionization source with positive ion mode detection. The linear range of the target compound was 0.7~104 μg/kg, the limit of detection (LOD) was 0.11~0.37 μg/kg, the limit of quantification (LOQ) was 0.36~1.22 μg/kg, the recovery rate was 84.0~116.3%, and the relative standard deviation (RSD%) was 0.8~10.5. The samples of maize plumule, brassica rapeseed flower, and marigold leaf were detected using the external standard method. The optimization of the extraction method and detection method of EBL improved the detection sensitivity, laid a foundation for the artificial synthesis of EBL, improved the extraction rate of EBL, and provided a theoretical basis for the study of EBL in many plants.

This study evaluated the effects of different temperatures (5 °C, 10 °C, 15 °C, and 25 °C) on the f

Ensiling forage under low-temperature conditions often leads to poor fermentation and nutrient losses. This study evaluated the effects of a cold-tolerant Pediococcus pentosaceus OL77 strain on oat silage. Silages were prepared with or without Pediococcus pentosaceus inoculation (1 × 10⁵ cfu/g FM). After 90 days, OL77-treated silage showed markedly higher lactic acid (45.83 vs. 30.51 g/kg DM), lower pH (3.88 vs. 4.443), and better preservation of WSC (64.68 vs. 47.60 g/kg DM) and crude protein (89.26 vs. 65.52 g/kg DM) than the control. Microbial analysis revealed accelerated colonization by Pediococcus, reduced bacterial diversity, and faster stabilization of the fermentation process. Functional predictions indicated enhanced carbohydrate and energy metabolism. These findings demonstrate that OL77 can effectively improve fermentation quality and nutrient preservation of oat silage under low-temperature conditions, offering a practical inoculant option for cold regions.