Lai's Metarhizium anisopliae|354188 |BNCC

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Metarhizium rileyi

Subculture procedure	① Dissolve the frozen tube in a water bath (fungi at 30 ℃, bacteria at 37 ℃) and shake quickly for dissolution; ② Wipe the outer wall of the cryovial with an alcohol swab for disinfection, and then transfer it to a safe cabinet for operation; ③ Unscrew the tube cap, aspirate all the dissolved solution, and transfer it into 1-2 agar plates (each plate contains approximately 200 μL, and for anaerobic bacteria cultivation, the culture medium needs to be placed in an anaerobic environment for 24 hours to remove oxygen); ④ After spreading evenly, transfer to the above cultivation conditions for cultivation.
Growth conditions	28 ℃; 5-7 days; aerobic;
Storage conditions	2-8 ℃
Safety level	1
morphology	Small filamentous fungi have obvious colonies on comprehensive PDA medium, with white and dense hyphae in the early stage, and green spores growing all over the agar plate later
Sharing mode	Public welfare sharing

M. rileyi

Storage conditions: 2~8 ℃

No. 354188

Product format: freeze dried, 200ul

Validity : 6 years

Biosafety level: 1 , handle in ultra-clean 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 plate 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 ℃, aerobic, comprehensive PDA,5-7 days, comprehensive PDA: potato boil 1.0L, glucose 20.0g,KH2PO4 3.0g,MgSO4·7H2O 1.5g, vitamin B1 trace, agar 20.0g,pH 6.0±0.2. Sterilization at 121 ℃ for 15min. Potato boiling solution: weigh 200g of peeled potato pieces, boil in boiling water for 30min, and collect filtrate to a constant volume of 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 5-7 days.
Recovery record: According to the recovery instructions, the results of the recovery are reported as follows:

Item	test results
viability:	good viability， in 7days strain layer is obvious
colony morphology: (above)	small filamentous fungi have obvious colonies on comprehensive PDA medium, with white hyphae at the initial stage, dense and vigorous, and green spores growing on the plate later.
Conclusion:	good viability, no abnormal colony morphology, qualified

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