Eelworm killing metarhizium and application thereof
A technology for Metarhizium anisopliae and knot nematode, applied in the field of Metarhizium anisopliae, can solve the problems of loss of preparation vitality, difficult commercial production, poor strain colonization ability, etc., and achieves the effects of broad application prospect, good colonization effect and convenient application.
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Embodiment 1
[0039] Example 1. Isolation, Identification and Preservation of Metarhizium anisopliae var. anisopliae HYN-1-2 CGMCCNo.5433
[0040] Collect Hebei Yongnian root-knot nematode diseased root of cucumber, wash the diseased root with tap water for 3-5 minutes to remove the soil, then put the diseased root under the dissecting microscope, pick out the oocyst with small forceps, and treat the oocyst with 1% sodium hypochlorite for 1 minute on the surface of the oocyst Disinfect and release nematode eggs, pass the sodium hypochlorite egg suspension through 200 μm and 30 μm mesh sieves, rinse 3 times with sterilized distilled water to remove sodium hypochlorite, spread approximately every 100 eggs on a PDA plate, and observe the appearance of colonies every day . Finally, a fungal strain that kills nematodes was isolated from the eggs of root-knot nematode of cucumber, named HYN-1-2, and was deposited in the General Microbiology Center of China Committee for the Collection of Microbial ...
Embodiment 2
[0042] Embodiment 2, the cultivation of Metarhizium anisopliae var. anisopliae HYN-1-2
[0043] 1. Test tube culture
[0044] The hyphae or spores of Metarhizium anisopliae HYN-1-2 are inoculated on the slant of the test tube PDA medium, cultured at 25-27° C. for 72-96 hours, and the test tube species is obtained.
[0045] 2. Bacteria Shaker Expansion Cultivation
[0046] The slant seeds were inoculated into the liquid YES-2 medium in the Erlenmeyer flask, and cultured on a shaking table at 25-26° C. and 120-180 rpm.
[0047] 3. Liquid fermentation production
[0048] Inoculate the metarhizium anisopliae HYN-1-2 bacterium liquid that expands step by step in the fermenter (fermentation medium is YES-2 medium) according to the volume ratio of 1:10, stir and ventilate at 25-26 ℃ (air flow 1 : 1), cultivated for 48 hours, and the concentration of Metarhizium anisopliae HYN-1-2 was 4×10 9 -7×10 9 cfu / mL of fermentation broth.
[0049] The formula of the above-mentioned YES-2 ...
Embodiment 3
[0050] Example 3, Preparation of Microspherical Preparation of Metarhizium anisopliae var. anisopliae HYN-1-2
[0051] 1. Preparation of microsphere sodium alginate gel (100 liters): slowly add 2.6 kg of sodium alginate into 100 liters of tap water at 80°C, stir continuously to dissolve it, and obtain an aqueous solution of sodium alginate, which is cooled to about 30℃, standby;
[0052] 2. Get example 2 to obtain the concentration of Metarhizium anisopliae HYN-1-2 as 4×10 9 100 liters of cfu / mL fermented liquid was homogenized and crushed, and 24 kg of diatomaceous earth was added to mix well, then added to 100 liters of sodium alginate aqueous solution prepared in step 1, fully stirred and mixed to obtain a suspension;
[0053] 3. Preparation of CaCl 2 Solution: prepare 100 liters of calcium chloride solution with a molar concentration of 0.2M with tap water, and set aside;
[0054] 4. Pass the suspension obtained in step 2 through a sieve with an aperture of 0.2 mm, and ...
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