Four new polyketides, botryosphaerones A -D (1 -4, resp.), were obtained from the fermentation culture of Botryosphaeria australis strain ZJ12-1A, together with four known compounds, Omethylasparvenone (5), 6-ethyl-2,7-dimethoxyjuglon (6) and its monoacetyl derivative 7, and Omethylaspmenone (8). Their structures were elucidated by spectroscopic analyses, including 1D-and 2D-NMR experiments, and HR-Q-TOF mass spectrometry, and by comparison with reported data. All compounds were evaluated for their cytotoxic and antimicrobial activities in vitro. Only compounds 6 and 7 showed cytotoxic and antimicrobial activities, as already reported.Introduction. -Plant-associated fungi, including endophytic and epiphytic fungi, are important sources of bioactive compounds. Previously, we mainly focused on the bioactive metabolites from endophytic fungi [1 -5]. In our search for antitumor compounds from epiphytic fungi, we observed that the fermentation extracts of strain ZJ12-1 exhibited strong cytotoxic activity. This strain ZJ12-1A was originally obtained from the plant epidermis of Sonneratia apetala Buch. Ham (a kind of mangroves), and it was identified as Botryosphaeria sp. based on its complete ITS4-5.8S-ITS5 gene sequences. Various compounds were isolated from the genus Botryosphaeria. Some of them showed interesting biological activities; i.e., antifungal and cytotoxic depsidones [6], antimicrobe diterpenoids [7] and antiseptic mellein [8], antimicrobial lasiodiplodin [9], dihydronaphthalenones [10], and dihydrobenzofuran, and antibacterial primin [11]. The current study on the chemical constituents of strain ZJ12-1A resulted in the isolation of four new naphthalenones, named botryosphaerones A -D (1 -4, resp.), together with four known compounds 5 -8 (Fig. 1). Here, we report the isolation and structure elucidation of compounds 1 -4. The in vitro antimicrobial and cytotoxic testing of compounds 1 -8 are also described.