1,5-Dihydroxy-3-methoxy-7-methylanthracene-9,10- dione (shiraiarin) is a kind of antitumor and antibacterial anthraquinone, and was produced for the first time from the submerged fermentation of Shiraia bambusicola, as confirmed by ESI-MS and NMR. The production of shiraiarin was significantly influenced when varying the carbon source, and a high amount of shiraiarin was only achieved when using lactose. The production of shiraiarin was also stimulated when using NaNO3 as the nitrogen source, whereas other nitrogen sources inhibited its production. Shiraiarin was formed during the stationary phase with a pH value higher than 8. The production of shiraiarin was inhibited by sporulation.