(I) Strains Isolation
Common methods: Tissue separation, spore separation, basal mycelium separation.
Preferred method for Flammulina velutipes: Special pith separation (due to small, thin caps). Steps: Grasp the stem, remove the cap along the stem with tweezers to expose the curved growth point at the stem top; transfer tissue from this point to a slant culture medium. This method has a high success rate.
(II) Stock Culture Production
Key note: Strain quality directly affects yield. High-quality strains (in-house developed or imported) are selected. Pure strains from fruiting bodies must undergo fruiting tests before large-scale use to avoid losses.
Common slant culture medium formulas for mother cultures:
Potato dextrose medium (PDA): 200g peeled potatoes, 20g glucose, 18-20g agar, pH 6.5-7.0, 1000ml water.
Potato comprehensive culture medium: 200g peeled potatoes, 20g glucose, 18-20g agar, 3g potassium dihydrogen phosphate, 1.5g magnesium sulfate, pH 6.5-7.0, 1000ml water.
Enoki mushroom rejuvenation medium: 200g peeled potatoes, 20g glucose, 0.5g peptone (note: original "58%" likely a typo), 18-20g agar, 3g potassium dihydrogen phosphate, 1.5g magnesium sulfate, pH 6.5-7.0, 1000ml water.
Preparation of mother culture medium: Follow conventional methods.
Propagation of mother culture: Inoculate vigorous, uninfected mycelia into sterilized blank agar slant medium under aseptic conditions; culture at ~23°C. It becomes production-ready mother culture after 7-12 days.
Note: Mother cultures easily form fruiting bodies during transport, so production test tubes must be promptly expanded into stock cultures.
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