A kind of metarhizium anisopliae and its biological control method and application in the pupal stage of Spodoptera frugiperda
A technology of Spodoptera frugiperda and Metarhizium anisopliae, applied in the field of microorganisms
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Embodiment 1
[0038] Embodiment 1: Isolation and identification of Metarhizium anisopliae FJMR2 bacterial strain
[0039] Isolation, purification and pathogenicity determination of 1FJMR2 strain
[0040] Metarhizium anisopliae FJMR2 of the present invention was collected on August 6, 2019 from the larvae of the diseased Spodoptera moth in the corn planting area of Dongfeng Town, Jian'ou City, Fujian Province. Put the collected carcasses in a 5mL sterile EP tube and bring them back, and gently shake the carcasses on PDA medium (200g of potatoes, 20g of glucose, 20g of agar, 1000mL of distilled water) until some conidia fall on the medium , cultured in a biochemical incubator at 26°C, and after sporulation, the conidia were transferred to a new PDA medium for purification to obtain the FJMR2 strain.
[0041] The purified FJMR2 strain was cultivated in a biochemical incubator at 26°C with SMAY medium (40 g maltose, 10 g peptone, 2 g yeast extract, 20 g agar, and 1000 mL distilled water). T...
Embodiment 2
[0046] Example 2: Infection characteristics of Metarhizium anisopliae FJMR2 to larvae of Spodoptera frugiperda at different ages
[0047] a. Inoculate Metarhizium anisopliae FJMR2 on SMAY solid slant medium, culture at 26°C for 7-9 days, and collect spores after the culture;
[0048] b. Dip a small amount of spore powder with a hairbrush, and brush it on the body walls of young larvae (1-3 instar larvae) and advanced larvae (4-5 instar) Spodoptera frugiperda larvae respectively, cultivate them at 26°C for 7-12 days, and observe Metarhizium anisopliae FJMR2 Infection characteristics of Metarhizium anisopliae FJMR2 on different ages of Spodoptera frugiperda larvae.
[0049] Results: The young larvae were infected with Metarhizium anisopliae and died on the 4th day at the earliest. In the early stage of death, the surface of the larvae was covered with a thin layer of hyphae. In the later stage, the larvae shriveled and stiffened, and no spores appeared. The time of death was re...
Embodiment 3
[0051] Embodiment 3: The pathogenicity determination of FJMR2 bacterial strain to Spodoptera frugiperda pupae
[0052] 1 Determination of the pathogenicity of different FJMR2 spore concentrations on Spodoptera frugiperda pupae
[0053] The culture method of Example 2 was used to collect Metarhizium anisopliae FJMR2 spores. The spores of Metarhizium anisopliae FJMR2 were prepared with an aqueous solution with a sterile mass percentage of 0.05% Tween-80 to a concentration of 3.00×10 8 Spores / mL, 3.00×10 7 Spores / mL, 3.00×10 6 Spores / mL and 3.00×10 5 Spore / mL spore suspension, 0.05% Tween-80 sterile water was used as blank control. Healthy 1-day-old Spodoptera frugiperda pupae of uniform size were selected for inoculation by dipping method. Place the pupae for testing in the prepared spore suspensions of different concentrations and immerse them for 5 seconds, take them out and dry them naturally, then place them in an incubator at 26°C, with a relative humidity of 70%, and ...
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