Culture medium for quickly screening filamentous fungi for producing polyunsaturated fatty acid and application of culture medium
An unsaturated fatty acid and culture medium technology is applied in the field of rapid screening of medium for polyunsaturated fatty acid-producing filamentous fungi, which can solve problems such as unreported, and achieve breakthroughs in time-consuming and labor-intensive, improving screening efficiency, and simplifying the screening process. Effect
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Embodiment 1
[0036] Example 1 Establishment of efficient screening method for polyunsaturated fatty acid-producing filamentous fungi
[0037] (1) Selection of bacterial strain: laboratory wild-type oleaginous filamentous fungus Mortierella alpina (Mortierella alpinaATCC 32222);
[0038] (2) Establishment of the screening method: the screening medium was based on PDA medium, and Mortierella alpina ATCC 32222, an oil-producing filamentous fungus already in our laboratory, was used as the test bacteria. Among them, the composition of the screening medium is: glucose 20g / L, potato extract 4g / L, agar 20g / L, streptomycin 30mg / L, cerulenin 1×10 -7 mol / L, TTC added concentration 0.5-2.0g / L. Incubate in the dark at 28°C for 3-7 days.
[0039] (3) Primary screening of filamentous fungi producing polyunsaturated fatty acids: prepare fresh Mortierella alpina ATCC32222 spore liquid, carry out plate coating after appropriate gradient dilution with 0.9% normal saline (the composition of plate culture m...
Embodiment 2
[0048] Example 2: Screening of filamentous fungi producing polyunsaturated fatty acids in the natural environment
[0049] Soil samples were collected from grasslands in the Qilian Mountain pastoral area at the junction of Qinghai-Gansu, and the surface soil was removed with a cleaning tool shovel, and about 150g of soil at a distance of 5-10cm was collected and packed in a sterilized centrifuge tube.
[0050] Same as Example 1, by dilution coating method, using the addition of 1×10 of the present invention -7 mol / L cerulenin and 0.5g / L triphenyltetrazolium chloride (TTC) culture medium to isolate oleaginous filamentous fungi, and culture them in the dark. The color (red) is dark and light, and the strains with fast growth speed, large colony diameter and red colony are selected as the target bacteria, and the target bacteria are transferred to the PDA medium plate for purification by using the mycelium edge purification method, and the purified bacteria are slanted. Store at...
Embodiment 3
[0053] Example 3: Screening of filamentous fungi producing polyunsaturated fatty acids in the natural environment
[0054] Soil samples were collected from the forest land of the Inner Mongolia Agricultural University campus, and the surface soil was removed with a cleaning tool shovel, and about 150 g of soil at a distance of 5-10 cm was collected and packed in a sterilized centrifuge tube.
[0055] Same as Example 1, by dilution coating method, using the addition of 1 × 10 -7 mol / L cerulenin and 0.5g / L triphenyltetrazolium chloride (TTC) culture medium to isolate oleaginous filamentous fungi, and culture them in the dark. The color (red) is light or dark, and the dark red target bacteria are transferred to the PDA medium plate for purification using the hyphae edge purification method, and the purified bacteria are stored on a slant at 4°C. Co-isolated and purified 5 strains of filamentous fungi.
[0056] The 5 strains of filamentous fungi screened were cultured in shake f...
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