Microbes serve as the most important resource for drug discovery. During our screening for bioactive compounds from our natural products library, a pathogenic fungus, Microdochium majus strain 99049, from wheat was selected for further investigation. A new alkaloid named brocaeloid D (1), together with six previously characterized compounds (2-7) were identified. Compound 1 belongs to 4-oxoquinoline with C-2 reversed prenylation and a succinimide substructure. All the structures of these newly isolated compounds were determined by different means in spectroscopic experiments. The absolute configurations of 1 was further deduced from comparison of its CD spectrum with that of known compound 2. The bioactivities of these identified compounds were evaluated against several pathogenic microorganisms and cancer cell lines. Compounds 1-5 showed activity against HUH-7 human hepatoma cells with IC(50) values of 80 mug/mL. Compound 6 showed mild activity against HeLa cells (IC(50)鈥?鈥?1.9 mug/mL), weak anti-MTB activity (MIC鈥?鈥?0鈥?mug/mL), and moderate anti-MRSA activity (MIC鈥?鈥?5 mug/mL), and compound 7 showed weak anti-MRSA activity (MIC鈥?鈥?00 mug/mL). CI - (c) 2019 Production and hosting by Elsevier B.V. on behalf of KeAi Communications Co., Ltd.