Cathamus tinctorius, Coptis japonica and Asarum sieboldii were tested as substrate for the production of Flammulina velutipes. Among the C. tinctoris , C. japonica and A. sieboldii , C. tinctoris was the best substrate for the production of fruitbody. The effects of addition of C. tinctoris to sawdust substrate resulted in the increased mycelial growth on inoculum culture, 3.1% in ratio of fully culture and shorted one day in culture period. C. tinctoris was decreased 6.1% in ratio of fully culture, 11.0% in ratio of fruitbody productive culm. The addition of C.tinctoris, C.japonica to sawdust substrate increased 134.6%, 114.1% on the yield of the mushroom fruitbody respectively . But A. sieboldii decreased the mycelial growth and pineheading ratio delayed the production of fruitbody.