Grifola umbellate is a famous and expensive Chinese herb medicine and the main medicinal component is polysaccharide mainly produced by its mycelia. Effects of organic nitrogen and carbon resources on mycelial growth and polysaccharides production of a medicinal mushroom, G. umbellate were studied in the submerged culture system. Soybean meal was selected as the optimal organic nitrogen source for its significant promotion effects on mycelial growth and polysaccharides synthesis based on results of an orthogonal matrix experiment design. Glucose was proved to be a preferred carbon source by comparison with fructose and sucrose. The optimal dosages of soybean meal 5.63% (w/v) and glucose 4.33% (w/v) were obtained, respectively through the curvilinear regressions experiment. The productions of mycelial dry weight, exo-polysaccharide (EPS) and inner polysaccharide (IPS), in the optimal medium, were about 3.16, 1.48 and 2.78 folds higher than those in the basal medium, respectively and culture duration was shortened to half through optimizing the organic nitrogen and carbon sources. These results will be significant contribution to process biochemistry of medicinal fungus in the submerged culture system.