The invention discloses a method for rapidly extracting long-fragment DNA from fresh fungus mycelia. The method comprises the following steps: thallus preparation: extracting fresh cultured fungus mycelia, carrying out centrifugation to remove moisture, weighing the mycelia, and rapidly grinding the mycelia into powder in liquid nitrogen; cell lysis: adding a DNA extracting buffer solution, adding60 mu L of 20mg/mL proteinase K, uniformly mixing, and carrying out water bath at 65 DEG for 30 minutes; DNA extraction: adding 0.33mL of KAc, carrying out water bath for 20 minutes, carrying out extraction by virtue of a mixture of phenol, chloroform and isoamyl alcohol, carrying out centrifugation, extracting supernate, adding absolute ethyl alcohol for precipitation, standing at a low temperature for 30 minutes, carrying out centrifugation, and collecting precipitates; purification: adding RNaseA, processing at 37 DEG C for 30 minutes, respectively extracting by virtue of a mixture of phenol, chloroform and isoamylol in a ratio of 25 to 24 to 1 and a mixture of chloroform and isoamylol in a ratio of 24 to 1, and carrying out low-temperature centrifugation for 5 minutes; and extractingsupernate, adding absolute ethyl alcohol for precipitation, selecting precipitates, rinsing, carrying out air drying, dissolving the precipitates into a TE buffer solution, and carrying out preservation for later use. The DNA amount of filamentous fungi extracted by virtue of the method is increased twice, the concentration of the filamentous fungi reaches 277ng/mu L, the lengths of fragments aremore than or equal to 20000bps, and the filamentous fungi are not degraded and are high in purity.