The invention discloses a co-production medium, a preparation method and application thereof. The co-production medium includes the following raw materials: minced rice straw, citrus juice, peanut cake, potassium chloride, sodium nitrate, ammonium sulfate, sodium iodate, calcium nitrate, magnesium sulfate, zinc nitrate, manganese chloride, sodium borate, sodium molybdate, ferric nitrate, copper chloride, N-potassium lauroylglutamate, alkyl sodium sulfovinate, an acrylate conditioning agent, diethylaminoethyl glycolate, 6-BA, indolebutyric acid, naphthylacetic acid and water. The co-production medium is prepared by the steps of raw material weighing, mixing, pH adjustment and sterilization. The co-production medium provided by the invention can achieve two things at one stroke when used for cultivation of flammulina velutipes and anoectochilus roxburghii, and the obtained lentinus edodes and anoectochilus roxburghii have high yield and good quality.