To find the new materials for Flammulina velutiper cultivation and to increase the economic benefits of kenaf cultivation, its byproduct was used in the test of Flammulina velutiper cultivation in the study. The effect of moisture, pH, calcium salts, sugars and carbonitride on the Flammulina velutiper growth was measured by mycelial growth rate and biological efficiency. The optimal medium conditions of kenaf byproduct for eryngii cultivation were also determined. The results showed that the culture medium contained 50% kenaf byproduct, 30 nitrogen source , 67.5% moisture content, 0.5% calcium carbonate，0.5% calcium oxide and 1% sucrose, pH value of 6.5, the biological efficiency of Flammulina velutiper could reached up to 111.0%. The variety screening results showed that ‘Jinza 19’ grew orderly, and its biological efficiency could reach more than 106.7%. ‘Changbai 201’ had a short growing cycle and good appearance, and its biological efficiency colud reach to 101.6%. This study demonstrated that the kenaf byproduct should be a good raw material for Flammulina velutiper cultivation.