Metarhizium anisopliae|114445 |BNCC

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Metarhizium anisopliae (Metschn.) Sorokīn

Subculture procedure	① Prepare 1-2 plates as mentioned above; ② Open the safety cabinet, burn the top with an alcohol lamp, quickly drip sterile water to break it, and then use tweezers to crush it; ③ Take 0.5mL of sterile water and transfer it into a freeze-drying tube. After fully dissolving the bacterial powder, transfer it into a agar plate at a rate of 200 μ L per piece and apply evenly; ④ Place the petri dish under the above cultivation conditions for cultivation, and once the bacterial strain grows, it can be used.
Growth conditions	28 ℃; 7-10 days; aerobic
Storage conditions	2-8 ℃
Safety level	1
morphology	A small filamentous fungus， which formed distinct colonies on comprehensive PDA medium. The colonies were white with dense and vigorous mycelia， later producing light yellow-green spores， exhibiting spreading growth towards the agar plate edge， and showing a yellow reverse on the medium.
Sharing mode	Public welfare sharing

Metarhizium anisopliae

Storage conditions: 2~8 ℃

No. 114445

Product format： freeze dried, 200ul

Validity period: 6 years

Biosafety level: 1, handle in ultra-clear table or safety cabinet

Receiving notice: if any abnormality is found on the day of receipt, please contact the customer service within 24 hours. If it is overdue, it is deemed that the bacteria are well. The freeze dried culture shall be used up once and shall not be retained. The bacterial viability will resume after 1-2 generation of recovery and can be used normally. Handling instructions of agar slant and suspension liquid is enclosed. Please operate in strict accordance with this instruction, otherwise the replacement of bacteria are not be available in case of aberrant growth and loss of viability.

Growth conditions: 28°C, aerobic, integrated PDA, 7-10 days. Comprehensive PDA: Potato cooking liquid 1.0L, glucose 20.0g, KH2PO4 3.0g, MgSO4·7H2O 1.5g, trace amount of vitamin B1, agar 20.0g, pH 6.0±0.2. Sterilize at 121℃ for 15min. Potato cooking liquid: Weigh 200g of peeled potato pieces, boil in boiling water for 30min, collect the filtrate and dilute to 1.0L.

Recovery steps:

(1)Prepare 1-2 of above mentioned plates;

(2)Open it in the biosafety cabinet, sterilize of the ampoule, heat the tip in a flame, quickly drop sterile water to creak it, then break it with forceps;

(3)Draw 0.5ml of sterile water into the freeze dried ampoule, make it fully dissolved, and distribute the solution to the plates well in 200ul/plate;

(4) Put the plates under the above culture conditions for cultivation for 7-10 days.

Recovery record: According to the recovery instructions, the results of the recovery are reported as follows:

Item	test results
viability	good viability, in 7 days plate colony is obvious
colony morphology:	small filamentous fungi with obvious colonies on comprehensive PDA medium, white, The hyphae are dense and vigorous, and then produce light yellow spores, which spread and grow to the edge of the plate, and the back of the medium is yellow
Conclusion:	good viability, no abnormal colony morphology, qualified

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