Abstract: Preparation of cultivation medium with corn straw for planting of Stropharia rugosoannulata is investigated. Single factor experimental design was employed to study the effects of corn straw freshness, degree of crushing, treatment method of cultivation material and moisture content of cultivation medium on the growth of Stropharia rugosoannulata. Then uniform experimental design was employed to optimize the ratio of main ingredient (corn straw) to supplementary ingredient (rice husk, wood chip and bran). The results showed that there was no significant difference between fresh corn straw and dry corn straw on the growth of Stropharia rugosoannulata. The bioconversion ratio of cut-off corn straw was higher than that of crushed corn straw, the vegetative growth time of cut-off corn straw was no significant difference in that of crushed corn straw. There was no significant difference between stacking fermentation and no fermentation on the growth of Stropharia rugosoannulata. The moisture content of cultivation material has no significant influence on the growth of Stropharia rugosoannulata when it was between 65% and 70%. The bioconversion equation of Y=(0.2283+1.6970A+2.0280C-1.1132A2-10.7559C2)×100% was obtained by regression analysis. Rice husk ratio(A) and wood chips ratio (C) had significant influence on the equation, but the bran ratio (B) had no significant influence on the equationn. When the fresh or dry corn straw was cut-off, the ratio of corn straw to rice husk, wood chips, bran was 1:0.762:0.094:0.05, the moisture content was 65%-70%, fermentation time was 3 days, the bioconversion ratio was 97.5% and the fruiting time was average 48 days. This cultivation medium with corn straw is simple and suitable for growth of Stropharia rugosoannulata.