Batch cultural method of nematophagous fungus in laboratory
A nematode-eating fungus and a culture method technology, applied in the field of batch cultivation of strains laboratory for the biological control of animal parasitic nematode diseases, can solve the problems of not being able to meet the requirements of batch cultivation and low production of nematode-eating fungi
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Embodiment 1
[0014] (1) The nematode fungus-Duddingtonia flagrans (synonymous with Arthorbotiys flagrans) strains stored in a test tube slant at 4°C were transferred to 1% bran Plate culture medium, cultured in the dark at 26°C for 6 days, and the culture with mycelia covered with the plate was used as the primary seed. Wherein, the preparation method of 1% bran plate culture medium is: take by weighing 10g fresh bran and add distilled water 1000ml, boil on low heat for 1 hour, filter with gauze, after precipitation, get the supernatant and mix after adding 2g of agar powder per 100ml, Then divide it into Erlenmeyer flasks, and sterilize under high pressure at 121°C for 20 minutes.
[0015] (2) Use a sterile scalpel to cut off 2 squares of 4 mm × 4 mm from the culture containing the first-class seeds, transfer it to a Erlenmeyer flask containing 2% bran liquid medium, and cultivate it at 26 ° C, first 80 r / Min shaking for 60 hours, then 160r / min shaking culture for 72 hours, the liquid ...
Embodiment 2
[0018] (1) Transfer the Arthorbotiys oligospora strains stored in the slant of the test tube at 4°C to 1% bran plate medium, and culture them in the dark at 25°C for 4 days, and the mycelia covered the plate. Two thirds of the later cultures were used as primary seeds.
[0019] (2) Use a sterile scalpel to cut off 3 squares of 4 mm × 4 mm from the culture containing the first-class seeds, transfer it to a Erlenmeyer flask containing 2% bran liquid medium, and cultivate it at 25 ° C, first 80 r / Min shaking for 48 hours, then 160r / min shaking culture for 72 hours, this liquid culture as the secondary seed.
[0020] (3) Pack corn grain and wheat grain compound culture medium into polyethylene plastic bags with a length of 37 cm and a width of 11 cm. One side of the plastic bag is equipped with a sterilizing filter membrane with a diameter of 5 cm to facilitate ventilation, and it is autoclaved at 121 ° C for 30 Minutes, after cooling, inoculate the secondary seeds into the abo...
Embodiment 3
[0022] (1) Transfer the strains of Pochonia chlamydosporia (Pochonia chlamydosporia) stored on the slant of the test tube at 4°C to 1% bran plate medium, culture them in the dark at 28°C for 7 days, and the mycelium grows to Two-thirds of the culture on the plate was used as primary seed.
[0023] (2) Use a sterile scalpel to cut off 3 squares of 4 mm × 4 mm from the culture containing the first-class seeds, transfer it to a Erlenmeyer flask containing 2% bran liquid medium, and cultivate it at 28 ° C, first 80 r / Min shaking for 72 hours, and then 160r / min shaking culture for 72 hours, the liquid culture was used as the secondary seed.
[0024] (3) Pack corn grains and wheat grains compound culture medium into polyethylene plastic bags with a length of 37 cm and a width of 11 cm. One side of the plastic bag is equipped with a sterilizing filter membrane with a diameter of 5 cm to facilitate ventilation, and it is autoclaved at 121 ° C. After cooling for 30 minutes, inoculat...
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