Culture medium for increasing growth speed and improving stress resistance of Esteya vermicola and preparation method of culture medium
A growth rate, medium technology, applied in the direction of microorganism-based methods, biochemical equipment and methods, with the addition of compounds to stimulate growth, etc., can solve the problems affecting the industrial production yield of Esteyavermicola fungus, low sporulation rate, growth rate, etc. Slow speed and other problems, to achieve the effect of improving growth rate and stress resistance, improving drought stress resistance, and increasing growth rate
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Embodiment 1
[0022] A kind of preparation method of the culture medium that improves growth rate and stress resistance of endoparasitic fungus Esteya vermicola of pine wood nematode, concrete steps are as follows:
[0023] (1) Peel and chop the potatoes, add 1L of water, boil for 30 minutes, filter with gauze, and collect the filtrate;
[0024] (2) Add 20g of glucose, 20g of agar, 0.1g of glycine, 5g of calcium chloride, add distilled water, adjust the pH value to 7, and set the volume to 1L;
[0025] (3) Put the solution after constant volume at 121°C, 1.05×10 5 Autoclaved under Pa for 20min to obtain culture medium.
[0026] Pour the culture medium into a petri dish under aseptic conditions. After the culture medium in the petri dish solidifies, cover the sterilized cellophane with a diameter slightly smaller than that of the petri dish; then transfer the activated pine wood nematode endoparasitic fungus Esteya vermicola to On the medium covered with cellophane, 10 plates were inoculat...
Embodiment 2
[0035] According to the preparation method of the culture medium in Example 1 and Comparative Example 1, respectively, glycine, serine, L-methionine, and L-leucine were added to the culture medium, each 100 μg / ml, and no amino acid was added as a control group. After inoculation of the fungus, 200 μl of approximately 2000 B. xylophilus nematodes were added on top of the 8-day-grown colony. The crescent-shaped spores of the fungus Esteya vermicola adhere to the body surface of the nematode, mainly the head and tail, and pierce the body surface and muscle layer of the nematode, absorb nutrients and germinate hyphae to produce spores, and finally exhaust the nutrients of the nematode. Its lethal. Observe and count the adhesion rate of pine wood nematode under a microscope for 10 hours, observe and count the lethality rate of pine wood nematode in 28 hours, the results are shown in Table 2. Each group was subjected to 3 repeated experimental treatments.
[0036] Adhesion rate an...
Embodiment 3
[0040]According to the preparation method of the culture medium in Example 1 and Comparative Example 1, respectively, glycine, serine, L-methionine, and L-leucine were added to the culture medium, each 100 μg / ml, and no amino acid was added as a control group. Eight days after fungal inoculation, spores of Esteya vermicola colonies were collected. Add 200 μl of the collected spore suspension cultured in different amino acid media to the bottom of the petri dish evenly. Expose at 25°C for 0.5, 1, 8, and 24 hours, respectively. Then add 1ml of sterile water and stir the spores with a glass rod. Aspirate 200 μl of spore suspension, add it to water agar, and test the germination rate of spores after culturing for 24 hours.
[0041] The Esteya vermicola fungus was transferred to the medium added with four different amino acids, and after culturing for 8 days, the spores of the Esteya vermicola colony were collected. Esteya vermicola spores were spread on water agar medium. The ...
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