To explore the chemical diversity of metabolites from new species of Dothideomycetes, the ex-type strain of <i>Sparticola junci</i> was investigated. Seven highly oxygenated and functionalized spirodioxynaphthalene natural products incorporating carboxyalkylidene-cyclopentanoid (<b>1</b>-<b>4</b>), carboxyl-functionalized oxabicyclo[3.3.0]octane (<b>5</b>-<b>6</b>), and annelated 2-cyclopentenone/δ-lactone (<b>7</b>) units, sparticolins A-G, were isolated from submerged cultures of the fungus. Their chemical structures including their relative (and absolute) configurations were established through spectroscopic and X-ray crystallographic analyses. Sparticolin B (<b>2</b>) exhibited inhibitory activity against the Gram-positive bacteria <i>Bacillus subtilis</i>, <i>Micrococcus luteus</i>, and <i>Staphylococcus aureus</i>, while sparticolin G (<b>7</b>) showed antifungal activities against <i>Schizosaccharomyces pombe</i> and <i>Mucor hiemalis</i>. All other sparticolins were only weakly active against <i>S. aureus</i> and also showed weak activities against the nematode <i>Caenorhabditis elegans</i>. Compounds <b>2</b> and <b>7</b> also showed moderate cytotoxic activities against seven mammalian cell lines.