Metarhizium anisopliae strain and application thereof
A technology of Metarhizium anisopliae and Metarhizium anisopliae, applied in the fields of application, fungi, and treatment of microorganisms with electricity/wave energy, can solve the problems of human and animal health hazards, environmental pollution, etc., and achieve the effect of strong insecticidal ability
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Embodiment 1
[0027] A strain of Metarhizium anisopliae of the present invention, the strain is Metarhizium anisopliae JXAS-03; the preservation name is Metarhizium anisopliae JXAS-03; it is preserved in China Typical Culture Collection Center, and the preservation address is Wuhan City, Hubei Province China Type Culture Collection Center of Wuhan University, Bayi Road, Hongshan District; date of preservation: August 14, 2019; preservation number: CCTCC M 2019626.
[0028] Metarhizium anisopliae JXAS-03 on the PDA plate, the initial vegetative growth stage is white, small filamentous fungus, grows radially outward, and the fungus becomes green or yellow-green in the later stage of sporulation, and the center of the colony is occasionally dotted of white.
[0029] The nucleotide sequence of the described Metarhizium anisopliae is shown in SEQ ID NO:1.
[0030] The Metarhizium anisopliae bacterial agent prepared by the Metarhizium anisopliae of the present invention. Its active ingredient i...
Embodiment 2
[0040] The difference between embodiment 2 and embodiment 1 is: the preparation method of scarab metarhizium anisopliae bacterial agent of the present invention, comprises the steps:
[0041] In step (1), single colony of bacterial strain is placed on the PDA plate containing the chloramphenicol of 25 μ g / ml, and temperature is 25 ℃, culture 14 days, until produce green spore, wash spore with 0.05% Tween-80, make Obtain Metarhizium anisopliae spore suspension bacterial agent;
[0042] In step (2), or pick a single colony in (1), inoculate it into LB culture solution containing 25 μg / ml chloramphenicol, at a temperature of 25°C, at a stirring rate of 100 rpm, and culture on a shaker for 3 days , preparing the fermentation culture of the Metarhizium anisopliae;
[0043] In step (3), after the fermentation culture in (2) is ultrasonically disrupted, the lysate is centrifuged at 3000 rpm / min, and the precipitate is the metarhizium anisopliae cell ultrasonic lysis precipitate; the...
Embodiment 3
[0045] The difference between embodiment 3 and embodiment 1 is: the preparation method of scarab metarhizium anisopliae bacterial agent of the present invention, comprises the steps:
[0046] In step (1), single colonies of the bacterial strain are placed on a PDA plate containing 25 μg / ml of chloramphenicol, and the temperature is 28° C., cultivated for 10 days until green spores are produced, and the spores are washed with 0.05% Tween-80 to prepare Obtain Metarhizium anisopliae spore suspension bacterial agent;
[0047] In step (2), or pick a single colony in (1), inoculate it into LB culture solution containing 25 μg / ml chloramphenicol, at a temperature of 28°C, at a stirring rate of 150 rpm, and culture on a shaker for 8 days , preparing the fermentation culture of the Metarhizium anisopliae;
[0048] In step (3), after the fermentation culture in (2) is sonicated, the lysate is centrifuged at 4000rpm / min, and the precipitate is the metarhizium anisopliae cell ultrasonic ...
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