Metarhizium anisopliae mfyy090714 and its application
A technology of MFYY090714, Metarhizium anisopliae, applied in the application, fungi, biocide and other directions, to achieve the effect of large spore production, simple culture, and difficult to produce drug resistance
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Embodiment 1
[0009] Embodiment one, the isolation of pathogenic bacteria, identification
[0010] 1. Materials and methods
[0011] 1.1 Materials
[0012] Adults of the diseased white-backed planthopper Sogatella furcifera (Horváth) infected by an entomophytic fungus were collected from terraced paddy fields in Yuanyang, Yunnan Province.
[0013] Potato dextrose (PDA) medium: 200g of peeled potatoes, cut into small pieces, add 1L of water, filter with gauze after 20min, add 20g of glucose and 15-20g of agar to the filtrate, heat to dissolve, and set to 1000mL Aseptic operating conditions: all All the utensils and utensils were sterilized in a high-temperature autoclave (121°C, 30min), and the inoculation and other operations were all carried out in an ultra-clean workbench.
[0014] Culture conditions: Culture in a 26°C light (12L:12D) incubator. After the colonies are formed, transfer to the test tube PDA slope, cultivate for 3 to 4 days, and transfer to a 4°C refrigerator for storage. ...
Embodiment 2
[0025] Embodiment two, biological characteristics of metarhizium anisopliae purified strain
[0026] 1. Materials and methods
[0027] 1.1 The strains to be tested Select a plate that grows uniformly and vigorously after purification as the strains to be tested. The hyphae were inoculated on PDA medium again, and cultivated in a constant temperature light (12L:12D) incubator at 26°C.
[0028] 1.2 Determination of colony growth rate and spore production. Take a plate of Metarhizium anisopliae that has been cultured in advance and use an 8mm puncher to punch a hole to get the bacteria, inoculate it on a new medium, do three repetitions, and place it at 26°C Cultured in a constant temperature and light (12D:12L) incubator, the diameter was regularly measured and recorded every day until the colony was full of the culture medium. Use a hole puncher with a diameter of 8mm to take the bacterium cake at the same position of the culture medium, add 1% Tween-80 and 20ml sterile water...
Embodiment 3
[0039] Example 3. Indoor pathogenicity determination of purified strains of Metarhizium anisopliae to white-backed planthopper
[0040] 1. Materials and methods
[0041]1.1 Source of tested insects
[0042] Select white-backed planthopper adults of the same batch, disease-free, and of the same size 24 hours after eclosion as test insects, and place the tested white-backed planthopper adults on rice seedlings planted in insect cages at 26±1°C and 12L: reared under the photoperiod conditions of 12D.
[0043] 1.2 Preparation of spore suspension
[0044] Get the purifying Metarhizium anisopliae that grow well and wash the conidia with sterile water and filter to prepare 10 8 Spores / ml conidia suspension, use a sterile capillary pipette to take a drop of the filtrate onto a hemocytometer, count the spores under a microscope and record the data, and dilute to 10 with sterile water containing 0.5% Tween 80 8 、10 7 、10 6 、10 5 、10 4 The spore suspensions with five concentratio...
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